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QUARTERLY JOURNAL 
OF EXPERIMENTAL PHYSIOLOGY 


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OUARTERLY 
POURNAL OF 
EXPERIMENTAL 
may SIOLOGY 


LONDON : 


EDITORS 

E. A. SCHAFER, EDINBURGH 
W. D. HALLIBURTON, LONDON 
C. S. SHERRINGTON, OXFORD 
E. H. STARLING, LONDON 

A. D. WALLER, LONDON 


VOLUME XI. 


pit. 
ipa eA 


CHARLES GRIFFIN AND COMPANY, LIMITED 
EXETER STREET, STRAND 


LOL 


PRINTED IN GREAT BRITAIN BY 
NEILL AND CO., LTD., 
EDINBURGH. 


7 g 
= 


CONTENTS OF VOL. XI. 


M. IraGaki, The Influence of Corpus luteum Extracts upon Plain Muscle, 
especially that of the Uterus. With Appendix by W. W. Taylor. 
(From the Department of Physiology, Edinburgh University.) With 
14 figures in the text 


M, Iracaki. On the Activn of various Extracts obtained from the Cow’s 
Ovaries upon the Muscular Tissue of the Uterus, Intestine, and Blood- 
Vessels. (From the Laboratory of Physiology, Edinburgh University.) 
With 12 figures in the text 


M. Iracaki. The Action of certain Gland Extracts and Drugs upon the 
Uterus of the Rat. (From the Laboratory of Physiology, Edinburgh 
University.) With 4 figures in the text 


P. T. Herrinc. The Effect of Thyroid-Feeding on the Weight of the 
Suprarenals and on their Adrenalin Content. (From the Physiology 
Department, University of St Andrews) . 


W. Cramer and R. M‘Caty. Carbohydrate Metabolism in Relation to the 
Thyroid Gland :—II.: The Effect of Thyroid-Feeding on the Gaseous 
Metabolism. (From the Imperial Cancer Research Fund, London, 
and from the Physiology Department, Edinburgh University.) Wath 
11 figures 


W. Brain Bett. Experimental Operations on the Pituitary. With 57 
tllustrations in the text 


J. AreyLu CamppeLt, Singapore. The Chemistry of Fossil Bone. (From 
the Departments of Physiology of the University of Edinburgh and 
of the Medical College, Singapore) 


A. S. F. Leyron and C. S. SHERRINGTON. Observations on the Excitable 
Cortex of the Chimpanzee, Orang-Utan, and Gorilla. With 30 figures 
in the text . . 


W.H. Tuompson. The Metabolism of Voluntary Muscle :—I.: The Effect 
of Prolonged Excitation of Motor Nerves on the Creatine Content of 
Limb Muscles. (From the Physiology Laboratory, Trinity College, 
Dublin) . . ; : : 


PAGE 


39 


47 


127 


135 


223 


v1 


P. T. Herrinc. The Action of Thyroid upon the Growth of the Body and 
Organs of the White Rat. (From the Physiology Department of the 
University of St Andrews) : : : : 

Masanaru Kosma, Fleet Surgeon, Imperial Japanese Navy. Studies on 
the Endocrine Glands :—Paper I.: The Relations between the Pancreas 
and Thyroid and Parathyroid Glands. (From the Physiology Depart- 
ment of Edinburgh University.) With 29 figures in the text and 


2 coloured plates 


Masanaru Kosima, Fleet Surgeon, Imperial Japanese Navy. Studies on 
the Endocrine Glands:—II.: The Relations of the Pituitary Body with 
the Thyroid and Parathyroid and certain other Endocrine Organs in the 
Rat. (From the Physiology Department of Edinburgh University.) 
With 13 figures in the text and 1 colowred plate . 


Masanaru Kogima, Fleet Surgeon, Imperial Japanese Navy. Studies on 
the Endocrine Glands :—III.: The Effects on the Thyroid and Para- 
thyroid of the Rat of administering Thyroid Extract and certain other 
Autacoids and Salts. (From the Physiology Department of Edinburgh 
University.) Wath 5 figures in the text . 


Masanaru Kogima, Fleet Surgeon, Imperial Japanese Navy. Studies on the 
Endocrine Glands :—IV.: The Effects in the Dog upon the Remainder 
of the Thyroid and Parathyroid of Partial Removal of those Organs. 
(From the Physiology Department of Edinburgh University.) With 
2 figures in the text 


Masanaru Kogsima, Fleet Surgeon, Imperial Japanese Navy. Studies on 
the Endocrine Organs:—V.: Effects upon Metabolism of Castration, of 
Thyroidectomy, of Parathyroidectomy, and of Thyroid and Parathyroid 
Feeding. (From the Physiology Department of Edinburgh University.) 
With 13 tables and 14 charts in an Appendix 


hk. J. S. M‘Dowauyt. A Cross-Striated Mammalian Muscle Preparation. 


(From the Department of Physiology, Edinburgh University.) With 
9 figures in the tert : 


PAGE 


255 


319 


337 


347 


351 


38] 


ALPHABETICAL LIST OF AUTHORS 


ARGYLL CampBELL, J. The Chemistry of Fossil Bone 

Buair Bert, W. Experimental Operations on the Pituitary 

Cramer, W., and R. M‘Catt. Carbohydrate Metabolism in Relation to 
the Thyroid Gland:—lII.: The Effect of Thyroid-Feeding on the 
Gaseous Metabolism : 

Herrinc, P. T. The Effect of Thyroid-Feeding on the Weight of the 
Suprarenals and on their Adrenalin Content 

——, The Action of Thyroid upon the Growth of the Body and Organs of 
the White Rat . 

IraGaki, M. The Influence of Corpus luteum Extracts upon Plain Muscle, 
especially that of the Uterus 


— ., The Action of various Extracts obtained from the Cow’s Ovaries upon 
the Muscular Tissue of the Uterus, Intestine, and Blood-Vessels 


——, The Action of certain Gland Extracts and Drugs upon the Uterus of 


the Rat . 


Kosima, Masauaru. Studies on the Endocrine Glands:—Paper I.: The 
Relations between the Pancreas and Thyroid and Parathyroid Glands 


——., Studies on the Endocrine Glands :—I1.: The Relations of the Pituitary 
Body with the Thyroid and Parathyroid and certain other Endocrine 
Organs in the Rat 

——,, Studies on the Endocrine Glands :—II].: The Effects on the Thyroid 
and Parathyroid of the Rat of administering Thyroid Extract and 
certain other Autacoids and Salts ; 

—— , Studies on the Endocrine Glands :—IV.: The Effects in the Dog upon 
the Remainder of the Thyroid and Parathyroid of Partial Removal of 
those Organs 

—-, Studies on the Endocrine Glands :—V.: Effects upon Metabolism of 
Castration, of Thyroidectomy, of Parathyroidectomy, and of Thyroid 
and Parathyroid Feeding . : ; g 

Leyton, A. S. F., and C. S. Suerrineton. Observations on the Excitable 
Cortex of the Chimpanzee, Orang-Utan, and Gorilla 


337 


347 


351 


135 


Vili 


M‘Catt, R., and W. Cramer. Carbohydrate Metabolism in Relation to 
the Thyroid Gland :—II.: The Effect of Thyroid-Feeding on the Gaseous 
Metabolism 


M‘Dowatt, R. J. 8S. A Cross-Striated Mammalian Muscle Preparation 


~y 


SHERRINGTON, ©. S., and A. S. F. Leyron. Observations on the Excitable 
Cortex of the Chimpanzee, Orang-Utan, and Gorilla 


Tuompeson, W. H. The Metabolism of Voluntary Muscle :—I.: The Effect 
of Prolonged Excitation of Motor Nerves on the Creatine Content of 
Limb Muscles 


PAGER 


59 
381 


135 


223 


QUARTERLY JOURNAL 
OF EXPERIMENTAL PHYSIOLOGY 


THE INFLUENCE OF CORPUS LUTEUM EXTRACTS UPON 
PLAIN MUSCLE, ESPECIALLY THAT OF THE UTERUS. 
By M. Itacaki. With Appendix by W. W. Taytor. (From the 
Department of Physiology, Edinburgh University.) (With fourteen 
figures in the text.) 
(Received for publication 26th July 1916.) 


THAT the corpus luteum is a gland of internal secretion appears to have 
been first suggested by Prenant (1), who arrived at this conclusion from 
the morphological character of its cells. Prenant’s conclusion has been 
very generally accepted, and a number of suggestions have been made 
regarding the nature of the effects which its supposed internal secretion 
produces. The literature of the subject will be found in detail in 
Marshall’s Physiology of Reproduction; the most recent suggestions 
are also referred to in Sir Edward Schiafer’s work on The Endocrine 
Organs, pp. 145-148, and except so far as they bear upon present investi- 
gations need not be further mentioned in this paper. 

It seems somewhat strange that, although the action of extracts from 
various ductless glands upon the uterus, and especially that of the supra- 
renal and pituitary body, has been investigated by several observers, very 
few papers have dealt with the influence of extracts of corpus luteum upon 
the contractions of the uterus, although the functions of these organs 
are closely related. I have only been able to find references in papers 
by Stickel (2), Fuchs (8), Guggisberg (4), and Ott and Scott (5), 
and the results which they have arrived at have differed consider- 
ably. Ott and Scott state that corpus luteum extract causes an increase 
in the contractions of the uterus both in pregnant and non-pregnant 
animals (rabbits and cats). Stickel found that in the rabbit parturition 
was hastened by injection of corpus luteum extract. Guggisberg failed 
to confirm Stickel’s observation, and often obtained, on the other hand, 
inhibition of uterine contractions both in pregnant and in non-pregnant 
guinea-pigs. It seemed therefore desirable to submit the subject to renewed 
investigation, and accordingly at the suggestion of Professor Schafer I have 
undertaken a series of experiments with a view to determining what effect, if 
any, is prcduced by extracts of corpus luteum upon the uterus; incidentally 
making a certain number of observations upon the action of the same 
extracts on other muscular tissue, especially that of the intestine. 

VOL. XI., NO. 1.—1917. t 


2 Itagaki 


METHOD OF PREPARING THE EXTRACTS. 


My extracts were in all cases made from the ovaries of the cow and 
sheep, the corpora lutea being removed and prepared as soon as they were 
received from the slaughter-house, i.e. within an hour or two after the 
animals had been killed. Occasionally the ovaries were kept overnight 
in an ice-chest at a temperature of from 3° to 7°C. The method which I 
have used for making the extracts has been as follows :—The corpora lutea, 
were shelled out from the ovaries by the aid of two pairs of forceps; the 
capsule or theca of each corpus luteum was then peeled off, and the 
remainder of the gland was minced and thoroughly pounded in a mortar 
until it formed a nearly homogeneous paste. Part of the paste was. 
taken and weighed, mixed with a definite amount of Locke’s solution, and 
boiled for a few minutes. It was then rapidly cooled, filtered, and made 
up to the original volume by the addition of distilled water. The reaction 
of such an extract is generally found to be neutral. This may be spoken 
of as extract of fresh corpus luteum. 

The remainder of the corpus luteum paste was spread out on a clean 
glass plate into a thin layer. This was placed in an incubator at a 


temperature of 37° to 40°C. for some hours until completely dry. It was _ 


then scraped off, transferred to a mortar, ground to powder, and kept in a 
desiccator. Roughly, the fresh corpus luteum by the process of drying 
was reduced in weight to one-fifth; in other words, it originally con- 
tained about 80 per cent. of water. 


TABLE I. 
Weight 
Date. Samples. eas Pa | proportion 
(weight). (weight). fresh to dew. 
GENS ie 10°] 1:9 5°32 
15,1.15 ie 11°3 1:9 6:00 
22.1.15 TET 19°1 3°75 5'1 
29.1.15 Vi: 141 2°5 5°64 
8.2.15 Ve 14°] 3°55 39 
15.2.15 VI. 21°30) 4°7 4°532 
22.2.15 VII 25°4 4:95 501 
4.3.15 Vill 29°35 54 5°435 
| 18.38.15 IX. 15°7 3°05 5°01 
| 25.3.15 x 66 12 55 
| 2.4.15 xa: 21°8 4°] 5314 
Average 5:16 


A weighed portion of this dried powdered corpus luteum was takem 
when required, mixed with a definite amount of Locke’s solution, boiled 


and cooled. It was then filtered, and distilled water was added to make. 
the solution up to its original volume. The reaction of this solution was. 


a 


Influence of Corpus luteum Extracts upon Plain Muscle 3 


also generally neutral. It will be referred to as extract of dried corpus 
luteum. 


METHOD OF RECORDING THE CONTRACTIONS OF THE UTERUS. 


The method which I have used for recording the contractions is 
a modification of that used by Magnus (6) in his investigations into the 
movements of the plain muscular tissue of the alimentary canal. The 
method has since been applied to the uterus (cat, dog, rabbit, and man) 
by Kehrer (7) and others. The following are the details of the method 
as employed by me:—A glass tube, 10 cm. long and 1°25 cm. internal 


Fic. 1,.—Diagram of tube for immersion of excised tissue in Ringer’s fluid 
and for replacement of this fluid by extracts. 


diameter, corked at the bottom and open at the top, serves to hold the 
tissue under investigation. Two side tubes which are fused in the main 
tube serve as inlet and outlet respectively for the fluid in which the uterus 
is to be immersed. The shape and relative position of these tubes are 
shown in the accompanying figure (fig. 1). The outlet tube is somewhat 
larger in diameter than the inlet. By means of a finely drawn glass tube, 
which was allowed to dip from above into the main vessel, oxygen was 
supplied in fine bubbles to the immersion fluid. 

The piece of tissue taken (usually from 1 to 1'5 cm. long) was fixed at 
the lower end to a hooked wire passed through the cork. The upper end 
of the tissue was attached by a fine thread to a recording lever of the first 
kind arranged to write upon a vertical drum moved by an electromotor at 
a slow rate, a time-tracing in minutes being always recorded underneath 
the myographic curve. The whole apparatus was immersed nearly to the 
open upper end of the main tube in a glass beaker filled with warm water,, 


4, Itagaki 


and this again in a considerably larger vessel. The latter was kept at a 
uniform temperature, and besides containing the above-described apparatus 
it served for holding small beakers containing different fluids for investiga- 
tion, which were therefore kept at the same temperature as that of the 
preparation upon which they were to act. 

The above modification of Magnus’ apparatus, which was devised by 
Professor Schafer, has two advantages over the original. In the first 
place, one is able to change the immersion fluid without emptying the 
tube in which the tissue is placed, so that the latter is completely sur- 
rounded by fluid during the whole of the experiment. There is further no 
risk of mechanically displacing the lever during the changing of the fluid, 
and the temperature of the fluid undergoes no alteration at any time. 

Mode of Preparation of the Tissue.—The animals from which the 
tissue was to be taken were always placed under chloroform. I found 
that the excised uterus from such animals nearly always shows fairly 
good contractions and regular rhythm. When completely anzsthetised 
the animal was bled to death by cutting through both carotid arteries. 
The abdomen was then opened freely, the intestine moved to one side, and 
the uterus found. The vagina having been cut through, the cut end was 
held by forceps and the cornua uteri separated from the adjoining 
structures. The whole of the uterus was in this way removed from the 
body, and was then placed in cold Ringer solution. The Ringer which 
I have employed throughout my experiments was prepared from Locke's 
formula, and was usually that furnished by Parke, Davis & Co. The 
formula is as follows :— 


NaHCO, ; . 0-01 per cent. 
CaCl, ; : : ; 0024.2 
KCl : : : 0042S ee 
NaCl . : : 09 


No glucose was added to the solution. 

As a general rule, a piece of one of the cornua was cut out and used as 
a whole. The record was mainly one of the longitudinal fibres, for, as 
Kehrer (7) and others have pointed out, the longitudinal fibres in the 
uterus are much stronger than the circular and exhibit far more powerful 
contractions. When larger animals were used, the whole of the cornu was 
not taken, but only a longitudinal piece of the wall. In some experiments 
portions of the uterus were used which had been kept immersed in Locke’s 


solution in an ice-chamber at a temperature of from 3° to 7° C. for from one 
to three days. 


In all the animals which I have investigated the normal movements of 
the uterine musculature are of two kinds, viz.: (1) regularly recurring 
ryhthmic contractions and relaxations comparable to the “pendulum 


Influence of Corpus luteum Extracts upon Plain Muscle 5 
movements” of the intestine; and (2) alterations in the general c ndition 
of contraction of the tissue, ie. alterations in tone, these being either 1 
the direction of increase of contraction or of diminution. The greater 
number of my experiments were made upon the uterus of the rat, and 


these may therefore be considered first. 


EXPERIMENTS UPON THE RATS UTERUS. 


At the moment when the ordinary Locke-Ringer solution was replaced 
by that containing extract of corpus luteum the immersed portion ot 


Fic. 2.—Tracing of cornu of rat’s uterus showing effect of immersion in extract of corpus luteum 
of cow. The tracing shows increase of tone, with rhythmic movements the amplitude of 
which gradually increases, the tonic contraction remaining at about thesame level. The time 
markings in this and in all the succeeding figures are in one minute. 


uterus showed in the majority of cases marked increase of tone, having 
almost the appearance of tetanus. A typical instance of such a tracing 
is shown in fig. 2, in which it will be observed that the increase of tone 
soon begins to show rhythmic contractions and relaxations, the amplitude 
of which gradually increases, although the maximum tonic contraction 
remains at very much the same point. On substituting normal Locke 
solution the effect at once begins to pass off. 

Variations were observed in the effect of the extract upon the rhythmic 
contractions. Sometimes these remained at the same rate as in ordinary 
Locke solution; at other times they became more frequent. In some cases 
the uterus was quiescent before the action of the extract. Under these 
circumstances extract of corpus luteum usually stimulated the uterus to 


9 


spontaneous contraction. This is shown in fig. 8. An effect of corpus 


6 Itagaki 


luteum upon the excised organ could be determined even after the uterus 
had been kept as long as three days in the ice-chest, although under these 
circumstances the result is less marked (fig. 4). 


Fic, 3.—Quiescent uterus of rat stimulated to activity by immersion in extract of 
corpus luteum. 


~ 


Fic, 4.—Tracing from the cornu of a rat’s uterus which had been kept in an ice-chest for three 
days. Between the two marks extract of corpus luteum was acting upon the uterus, Notice 
its stimulant action. (The signals should be shifted a little to the left.) 


In rare cases the effect of Ringer extract of corpus luteum, whether 
fresh or dried, produced, instead of increase of tone and increased rapidity 
of the rhythmic contractions, the opposite effect, viz. diminution of tone, 
which might amount to complete loss of tone, and diminution in height, or 


ain Muscle 


Influence of Corpus luteum Extracts upon P 


YASTOTIIUIT SBA STLIOZT 


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8 Itagaki 


even abolition of the rhythmic movements. Such a result is shown 
in fig. 5. 

The existence of pregnancy does not appear to have any specific in- 
fluence on the effect produced by corpus luteum extract upon the uterus 
of the rat. Usually the effect is that of increase of tone and increase of 
rate of rhythm of the individual contractions, but in one case out of four 
which were investigated inhibition was produced instead of increase. 

Sixty-seven experiments were made in all on the rat's uterus. Of these 
sixty-three were from non-pregnant animals. In fifty-three cases an in- 
creased contraction was produced. In seven the result was inhibition. In 
the remaining three no perceptible change was caused. 


~XPERIMENTS UPON THE RABBIT’S UTERUS. 


As a general rule the cornu of the rabbit’s uterus is too stiff and thick 
to be used as a whole, and in these cases the record was made from a 
longitudinal strip of the wall. In seven cases the uterus was taken from 
non-pregnant animals, in three from pregnant rabbits. The usual result 


Fic. 6.—Rabbit-uterus, Showing increase of tone and increased rapidity of rhythmic 
contractions as the result of immersion in corpus luteum extract (5 per cent.) 


in both cases was increase of tone, sometimes accompanied by an increased 
rate of the rhythmic contractions. Fig. 6 may be given as an example of 
this result. Inhibition was evident in two eases, one non-pregnant and 
one pregnant. After two days in the ice-chest the ordinary result was 
still obtainable. A few experiments were made with the rabbit’s uterus in 
situ, an injection of extract being made into the jugular vein. This usually 


Influence of Corpus luteum Extracts upon Plain Muscel 9 


caused a contraction of tetanic character showing a definite increase of tone 
which lasted for a few minutes (fig. 7). 


Fic. 


orpus luteum extract upon the contraction of the rabbit's ut in situ): 
10 c.c. ‘of ne per cent, extract of dried corpus luteum was il intraven vusly. This pro- 
duced at first « contraction of a somewhat prolonged character 


EXPERIMENTS UPON THE CAt’s UTERUS. 

A few experiments were made with the cat, one upon a pregnant uterus, 
the rest non-pregnant. The usual result was as with the rat, increase of 
tone, both with the fresh uterus and with that which had been kept in the 
ice-chest, although to a much less extent in the latter case. The increase 
of tone might be accompanied by acceleration of the rhythmic movements. 
In one of the experiments there was diminution of tone. The increase of 
tone is shown in fig. 8; diminution in fig. 9. 


EXPERIMENTS UPON THE Doa’s UTERUS. 


Only one experiment was made; upon a bitch’s uterus. In this case 
there was both an increase of tone and an increased rate of frequency of 
the rhythmic contractions (fig. 10). 


EXPERIMENTS UPON THE UTERUS OF THE GUINEA-PIG. 


Four experiments were made on the guinea-pig, all on non-pregnant 
animals. The result was a very great increase of tone with a tetanic 
character of the rhythmic contractions, which showed almost complete 
fusion. On substituting ordinary Locke there was at once great diminu- 
tion of tone, followed by large rhythmic movements. 


EXPERIMENTS UPON PLAIN MUSCLE OTHER THAN THAT OF UTERUS. 


Intestine of Rat.—A number of experiments—ten in all—were 
made upon pieces of the jejunum of the rat removed from just below 
the duodenum and placed in Locke solution. The pieces were emptied of 
their contents, washed with Locke solution, tied at both ends with thread, 


Itagaki 


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"09 JO oORIJXA TINayNt snd.too9 “yua0 ted g. 


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Influence of Corpus luteum Extracts upon Plain Muscle l] 


and fixed in the recording apparatus. Three out of the ten experiment 
show diminution of tone and inhibition of the “pendulum movements 
One shows diminution of tone only. Of the remaining seven, one shows 


Fic. 9.—Cornu of cat-uterus (excised). Showing diminution of tone caused by corpus 
luteum extract of cow. The immersion in corpus luteum extract was during the 
period of time marked by the arrows. 


Fie. 10.—Cornu of bitch’s uterus, excised. Showing increase of tone and increased rate of 
frequency of the rhythmic contractions as the result of immersion in 5 per cent. corpus 
luteum extract. 


an initial contraction followed by diminution of tone, and this again by 
increase of tone. In the other six cases the results were very slight, or no 


change at all could be observed. 
Intestine of Rabbit.—Eight experiments were made upon portions 


taken from the upper part of the jejunum treated in the same way as 


12 Itagaki 


the intestine of the rat. Without exception these showed increase of tone, 
althouch occasionally this was preceded by a short period of relaxation. 


Fig. 11 shows a typical experiment. 


Fic, 11.—Excised portion of small intestine of rabbit (whole tube). Showing increase of tone 


caused by immersion in 5 per cent. corpus luteum extract. The extract was added at the 
point marked by the arrow. 


Fic. 12.—Longitudinal strip of small intestine of rabbit. Showing diminution or 
tone with cessation of the rhythmic movements as the result of immersion in 
5 per cent. corpus Inteum extract. 


Fic. 13.—Portion of colon of rabbit (whole tube). Showing diminution of tone and 
abolition of rhythmic movements caused by 2 per cent. corpus luteum extract. 


Influence of Corpus luteum Extracts upon Plain Muscle 13 


In other cases longitudinal strips of the small intestine were employed, six 
experiments being made. Except in one case, which showed contraction 
although this did not appear until six minutes after immersion in the fluid 
—the strip became relaxed, showing diminution of tone, sometimes with 
and sometimes without, cessation of the “ pendulum movements” (fig. 12). 

Three experiments were made upon the lower part of the colon of a 
rabbit, which was excised, washed with Locke, tied at both ends, and 
placed in the apparatus. In all of these diminution of tone was observed, 
usually with abolition of the “pendulum movements” (fig. 13). 


Pcl yl Muy aula han) 


Fic. 14.—Longitudinal strip of portion of intestine of cat: the intestine had been kept twenty- 
four hours in an ice-chest. Notice the great diminution of tone with arrest of the rhythmic 
movements as the result of immersion in 5 per cent. extract of fresh corpus luteum of cow 
during the period marked by the strokes on the signal line. (These strokes should be shifted 
a little to the left. ) 


Intestine of Cat.—KEleven experiments were made on the intestine 
of the cat, and one upon the jejunum of a young kitten. In the last 
case diminution of tone was observed. In the adult cat the whole 
intestine was not taken, but strips of the longitudinal muscular coat. Of 
the ten experiments performed in this way, six exhibited diminution of 
tone with cessation of “pendulum movements.” These effects were 
recovered from on substituting Locke solution. In two cases there was 
increase of tone along with increase in the rate of the individual contrac- 
tions. In the remaining two cases no change could be observed. 

One experiment was made with a longitudinal strip of a piece of cat’s 
intestine which had been kept for twenty-four hours in the ice-chest. In 
this case, great diminution of tone, with arrest of the “pendulum move- 
ments,” was produced (fig. 14). 


14 Itagaki 


In four other experiments in the cat, portions of the circular muscle 
of the small intestine were employed; three of these showed distinct 
diminution of tone; in the fourth no clear effect was observable. 

Bladder of Rabbit.—Three experiments were made with strips 
of the rabbit's bladder, the mucous membrane having been shaved off. 
In all three cases diminution of tone was observed. 

The Iris of the Frog.—The eyes of a frog which had just been 
killed were enucleated and immersed, the one in Locke solution and 
the other in Locke extract of corpus luteum, the diameters of the pupils 
being carefully measured from time to time. The following table is a 
record of two experiments :— 


TABLE II. 
A. 
Locke. Corpus luteum. 
Date: reine Diameter of pupil. Diameter. 
Long. | Short. Long. Short. 
3.5.15 1.43 28 | 21 2:8 v2 | 
3.11 2°7 271 2°8 2°2 
4.0 2°6 18 2°7 159 
5.30 2°3 14 2°3 15 
B. 
8.6.15 11.20 31 |) ORS an 
12.0 30 2°2 2°8 2°1 
2.5 | 2°9 | 2°0 DET) 19 
4.0 S65 le ey 2:5 17 


It will be observed that there is no perceptible effect produced by corpus 
luteum. 


EFFECTS ON BLOOD-PRESSURE AND ON VOLUME OF KIDNEY. 


A number of experiments were made to determine whether intravenous 
injection of the extract produces any definite effects upon blood-pressure 
and kidney volume, and also, in some experiments, on the flow of urine; 
in lactating animals the flow of milk was observed. Blood-pressure was 


taken from the carotid artery. 


TaBLE II].—Rapsesir. 


| No. | Date. | Dose. Blood-pressure. ae Remarks. 
volume. 
1 1212-15 |) 159%, Sic.e! Fall | 
2 29.12.15 NO = 55 - | Increase 550 
3 SOSA 5: a) beanies Nochange | No change in 
| pulse rate. 


Influence of Corpus luteum Extracts upon Plain Muscle 15 
TABLE IV.—Car. 
No Date. Dose. roe ae Remarks. 
—— a7 
1 30.3.15 56% 5c.c. | Fall No change | Lactatinganimal. Secretion | 
of milk produced. 
2 14.6.15 10% 10 c.c 7” m 
3 18.6.15 » Sc. | Very slight No change in respiration, | 
fall Contraction of uterus. 
4 24.6.15 * No change Increase s is 
5 | 25.6.15 Re * ze No change | No change in respiration. | 
6 28.6.15 5% 5c.c. Increase Lactatinganimal. Secretion 
of milk produced. 
7 | 15.7.15 | 10% 5c.c. : 
8 8.15 | » | Alittle fall Nochange No change in blood-pressure 
before and after cutting | 
| vagi. 
9 | 19.815 ‘ » | Nochange | Increase 
10 | 30.11.15 ° | ae | Fall 
a, 3.1215 | (Cy, “ a Decrease No change in pulse rate. 
12 i 6.12.15 ” ” ” ” ” ” 
13 | 93.1215 | » : és f ‘ 
14 | 24.12.15 | , 4 | Slightfall | 
| Jt a Se No change | 


Below (8) states that a small dose of corpus luteum extract causes. 
a fall of blood-pressure, and a slow pulse with strong heart-beat, whereas 
a larger dose causes a greater fall of blood-pressure, with acceleration of 
pulse and a smaller amplitude of cardiac contraction. He found, on the 
other hand, that extract of ovary without corpus luteum raises the blood- 
pressure and accelerates the pulse. Biedl (9) was unable to confirm 
these results. Schickele (10) described the juice obtained from corpus. 
luteum as causing a fall of blood-pressure, continuing for several 
minutes (dog and rabbit); but he also found that the juice from other 
glands (thyroid, thymus) causes a fall of blood-pressure. As Bied] 
points out, it must therefore be doubted whether Schickele’s results. 
are characteristic of corpus luteum. 

In my own experiments a dose of from 5 to 10 cc. of a 5 per 
cent. to 10 per cent. extract of corpus luteum was injected into the 
jugular vein. This comparatively large dose produced only a slight fall 
of blood-pressure—indeed in some cases it had no effect—nor was there 
any change in the pulse rate. This has also been noted by Ott and 
Scott (11). In some of my experiments the kidney volume was increased, 
in others it was decreased. In many there was nochange. There is there- 
fore no specific action upon the kidney. The specific action upon the 
mammary gland, which has been demonstrated by Ott and Scott (11) 
and by Schafer and Mackenzie (12), is confirmed by my experiments. 
The secretion of urine appears to be unaffected (see also Ott and 
Seott (5)). 


16 Itagaki 


DoEs CorPUS LUTEUM ACT THROUGH THE UTERINE NERVES OR 
DIRECTLY ON THE MuscuLAR TISSUE ? 


I have further made a number of experiments with the object of 
ascertaining on what part of the neuro-muscular mechanism of the 
uterus an extract of corpus luteum acts, i.e. whether it produces its 
effect by stimulating nerve-endings or whether it is a direct action upon 
the muscular tissue. These experiments consisted in the successive 
application of adrenalin solution and corpus luteum extract to the same 
uterus (rabbit), and in the successive application of adrenalin (Parke, 
Davis & Co.) and corpus luteum extract to the uterus of different kinds 
of animals. The results are given in Tables V., VI., VII, and VIII. 


TABLE V.—RaABBIT. 


| | 

| Adrenalin. | Corpus luteum. 

| No Date. |e 

| | Dose. | Dose. 

| 1.6.15 1 : 10,600 Increase of tone | 0°5% _ | Increase of tone. 

eee 15.6.15 | 3% suprarenal i. Boe a 

| | | gland extract | | 

| 3 | 5.7.15 | 1: 10,000 A . ‘ 

| 4 | 20.7.15 ” ” ” ” 
TaBLE VI.—Rar. 
Adrenalin. Corpus luteum. 

Now 4) Dates oy ; 5 

| Dose. | Dose. 

1 | 13.2.15 | 1:10,000 | Decrease of tone and in- 5% Increase of tone 
| | hibition of rhythmic and rate of 
movements. rhythm. 

2 | 14.5.15 | ‘ f : 

3 | 25.6.15 % Wee «| ‘ 

Taste VII.—Cat. 
Adrenalin. Corpus luteum. 
No Date. =a 
Dose. Dose. 

A asa 1:10,000 | Decrease of tone and in- | 5% Increase of tone 
hibition of rhythmic | and rate sof 
movements. | rhythm. 

2 8.6.15 ” x”) | re) 9 

3 | 24.615 fi . ie | - 


Influence of Corpus luteum Extracts upon Plain Muscle 17 


TasBLe VIII.—GuInea-Pia. 


Adrenalin, Corpus luteum. 
| No. | Date. 
) / ' Dose. Dose. 

mi 67.616 1 : 10,000 Decrease of tone and in- 57 Increase of tone 
hibition of rhythmic and rate of 
movements rhythm, 

2 11.6.15 | 3% suprarenal | 
| gland extract 
Pg 4.9.15 | 1:10,000 
cee) 686 4.9.15 | 2 


Langley and Anderson (13) showed that stimulation of the hypogastric 
produces certain effects upon the uterus in the rabbit, but they failed to 
get any result on stimulating the pelvic nerve. They concluded that the 
hypogastric is the only nerve in the rabbit which influences uterine con- 
traction; and as corpus luteum generally produces the same effect in this 
animal as adrenalin, which is known to operate through the sympathetic 
nerve-endings, the conclusion is that the action is not through the pelvic 
nerve, although it may be through the hypogastric (sympathetic). 

The effects produced by corpus luteum upon the uterus of the rat, cat, 
and guinea-pig are the reverse of those which are produced by adrenalin ; 
it therefore appears clear that the effect obtained is not that of the 
sympathetic, although James and John Gunn (14) are of opinion that 
at present there is no evidence that the sympathetic in the guinea- 
pig and rat has any motor fibres for the uterus. The result of my own 
experiments, although showing that the action of corpus luteum extract 
is not equivalent to that of adrenalin, affords no proof that it influences the 
uterus through the mediation of sympathetic nerve fibres. Whilst, there- 
fore, it is possible that the effect may be a direct one upon the muscular 
fibres, I have not succeeded in devising experiments which are able to 
determine this point. 


EFFECT UPON THE EXCITABILITY OF THE VAGUS NERVE. 


I have made a few experiments to determine whether extract of corpus 
luteum affects the excitability of either efferent or afferent fibres in the 
vagus. Villémin (15) describes inexcitability of the vagus following 
injection of corpus luteum extract, and his experiments were apparently 
confirmed by Busquet (16). The latter, however, on reinvestigating the 
subject along with Pachon, obtained uncertain results, and came to the 
conclusion that there was no constant action upon the vagus nerve. In 
my experiments the effects on carotid blood-pressure were used as an 
indicator. In this way it was possible to observe the slowing of heart 

VOL. XI. NO. 1.—1917. 2 


18 Itagaki 


on stimulating the distal cut end of one vagus and, after section of the 
opposite vagus, the depressor action on stimulating the central end, 
and to endeavour to determine whether these results were influenced 
by intravenous injection of extract. Twenty experiments were made 
altogether, ten on rabbits and ten on cats. The results obtained were 
inconstant. 


CAUSE OF THE DIFFERENCES OF RESULT PRODUCED IN DIFFERENT 
EXPERIMENTS UPON THE UTERUS BY CORPUS LUTEUM. 


I have endeavoured to determine the cause of the differences of result, 
such as increase of tone in one case and diminution of tone in another, 
which are produced upon the uterus by extracts of corpus luteum. 

I first investigated the effects produced by different strengths of 
an extract. Within the limits of the dilution employed (0:1 per cent. to 
5 per cent.), the action was similar in each case. These results are 
exhibited in the following table. The numbers indicate the height of the 
ordinates in millimetres :— 


TaBLE IX. 


| Magnification 2 times. 


7.6.15 8.6.15 8.6.15 16.12.14 
1 2 3 4 5 


| Uterine contraction in mm. 
| 
| 
| 
| 
| 


o 
— 
[S) 


03 ys fit ae 

05 oe 10 ase ine 
0°65 5e6 ete cists ae 2 
1:0 13 isis 8 12 o0¢ 
1°25 BoE ase apo 4 
2°0 13°5 13 75 Soe 368 
2°5 565 ee er 11 8 
5 16 12°5 8 12 13 


This set of experiments shows clearly that alteration in the strength 
of the dose does not necessarily produce a difference in effect. It is note- 
worthy, however, that whereas in most experiments the effect was in no 
way proportional to the increase of dose, in one experiment—No. 5—there 
appeared to be an almost exact proportion between the effect in pro- 
ducing contraction and the strength of the dose. I am quite unable to 
explain this difference. Another circumstance which occurred to me as 
possibly influencing the result was the condition of pregnancy or non- 
pregnancy. But it will be seen from what has been already stated that 
this has no definite influence, for we find an inhibitory effect produced in 
both the pregnant and the non-pregnant uterus. That the effect is 
one of the extract and not of the condition of the uterus is shown 
in the following tables (X., XI, and XII.), which represent experiments 


Influence of Corpus luteum Extracts upon Plain Muscle 19 


upon the uterus of different animals with the employment of the 
same extract :— 


Taste X.—Corpus tureum Extract I, 


No. Date. Dose. 


l 30.12.14 | Rat uterus (a) 2 | Inhibition. 
2 31.12.14 | - (b) nat - 
3 31.12.14 ot) be RON ae m 


TaBLE XI.—Corpus Lureum Extract II. 


24.3.15 


No. Date. Dose. | | 
bya 24.3.15 | Rat uterus (a) | 25% Inhibition. 
2 ” (b) ” ” 


TaBLE XII.—Corpus LureumM Extract III. 


No. Date. Dose. | 
1 18.12.15 Rat uterus 5% | Inhibition. 
2 18.12.15 Rabbit uterus | : 


Further, when the same uterus is used, different extracts may give contrary 
results. This is shown in Table XIII. :— 


TABLE XIII. 


Effects of different extracts upon the same uterus. 


Date. == = aaa 
1g 2. | 3. 4, 


Increase of | Increase of 
tone tone. 


18.12.15 Inhibition and Increase of 
relaxation tone 


It is clear, then, that the differences of response of the uterus to corpus 
luteum extract are not due to differences in the condition of the uterus 
but to differences in the samples of extract used for investigation. 


In order to determine whether these differences were due to the 
condition of development of. the corpora lutea which were used for obtain- 
ing the extract, I made extracts from corpora lutea of different ages, 
i.e. of different degrees of development. I thought it not improbable that, 
since most of the corpora lutea which we employed were selected on account 
of their being in an advanced condition ofdevelopment, it might be possible 
to obtain a different result from extracts made from other corpora lutea 
which were less developed. With a view to test this notion, I made a 
series of experiments with extracts from corpora lutea showing different 


20 Itagaki 


appearances. The results were as follows:—(1) As a general rule the 
increase of tone was greater in extracts made from big corpora lutea, 
especially those having marked orange colouring. (2) As a general rule 
the extracts from large corpora lutea with yellow colouring also produced 
increase of tone. (3) Far less contraction was produced by extracts made 
from small corpora lutea with yellow colouring. Of fifty corpora lutea 
which were separately investigated, thirty-three of which belonged to the 
larger class, and the rest to the smaller, most of the first caused marked 
increase of tone. Of those which caused no marked increase of tone, 
seventeen were small yellow corpora lutea with thick theca. Amongst 
these, the extract of one caused inhibition and decrease of tone instead of 
increase of tone. Of the thirty-three extracts obtained from the large 
corpora lutea, six produced no marked change in uterine contraction. 

Experiments were also made to determine whether differences of effect 
were obtainable from corpora lutea of pregnant animals as compared with 
those of non-pregnant animals. By far the greater number of the corpora 
lutea which I have used were from the cow, and these were always non- 
pregnant animals. Since the corpus luteum of the pregnant cow was not 
obtainable, the corpus luteum of pregnant sheep was used in order to 
determine if the difference of the result were due to pregnancy. The ex- 
periments show that there was no difference in this relation. 


TaBLE XIV. 
No. of | | é 

extracts, | Date. Dose. 
| 1 9.3.16 5% Rat uterus. No difference. 
| 2 9 39 3? 

3 3° . bb) 29 ” 

4 23.3.16 if . 4 

o ” ’ > 


CHEMICAL INVESTIGATION. 


It appears therefore probable that there are two antagonistic principles 
present in the corpus luteum, both of which can be extracted with Locke 
fluid, although in the great majority of cases the principle which pro- 
duces inhibition is in far less amount or is absent altogether. I have 
made a series of attempts to separate these supposed antagonistic principles. 

I first tried the effect of extracting with alcohol and employing Locke 
extract of the dried alcohol extract. The method of extracting was as 
follows:—The corpora lutea were minced, ground in a mortar, and spread 
on a clean glass plate to dry, in the manner previously detailed. Over 
some of the dried powder cold absolute alcohol was poured, shaken several 
times, and the yellowish supernatant fluid when clear was decanted off and 
filtered. This procedure was repeated with successive portions of alcohol 


Influence of Corpus luteum Extracts upon Plain Muscle 21 


until the supernatant fluid showed no appreciable yellow colouring. The 
solutions thus obtained were mixed, evaporated to dryness over a water- 
bath, and an extract made from the dried residue by boiling for a few 
minutes with Locke solution. The extract was filtered. The residue of 
corpus luteum, after extraction with alcohol, was dried in an incubator at 
37° C., the fats and lipoids being in some cases first removed by chloro- 
form; occasionally Soxhlet’s extraction apparatus was used. The results 
of the testing of the various extracts upon the uterus are shown in the 


following tables (XV. and XVI.) :— 


TaBLE XV. 
ale ae | a | Solution. Residue. 
l 29.5.15 | Ratuterus 1% Inhibition | 1% Increase of tone. 
S 1.6.15 ” ” ” ” 
30.7.15 ks ce A : z 
s 01% Slight diminution 
| of contraction 
4 7.9.15 = 1% Inhibition 1% Increase of tone. 
8.9.15 | Rabbituterus _,, i _ ; “ 
(pregnant) 
9.9.15 | Cat uterus sy FP », Concentration. 
5 28.9.15 | Rat uterus No change 1% Increase of tone. 
Guinea-pig | 1% Increase of tone % S “s 
uterus 
6 29.9.15 | Rat uterus »  Nochange 
30.9.15 3 ree 1% Increase of tone. 
15.10.15 * Same extract as 29.9.15, | 
but 2°07%. Diminu- 
tion of tone. 
7 5.10.15 “ 1% No change 
10% Inhibition 
8 19.10.15 | Increase of tone 
9 | 8.10.15 | 5 1% Slowed rhythm 1% Increase of tone. 
coe)" 5.4.16 || : he 143% 3, ‘ 
1l LavG _ 1% Inhibition Il) SEPA - “ 
12 | 12.12.15 5 » Slight increase of 
tone 
13 4.4.16 58 _| 5% No change 2% Slight increase | 
| of tone. 
14 24.6.16 5 10%, 2%) 05% Diminu- | 1% Increase of tone. 
frequency 
15 20.6.16 : 10% Slight diminution > ¥3 a9 


of indiv. contr. 
5%, 10% No change 


16 21.6.16 ns 1% Diminution of », Slight increase of 


tion of tone and | 
indiv. contr. tone. 


22 Itagaki 


TABLE XVI.—ABSOLUTE ALCOHOL EXTRACTION AFTER EXTRACTION WITH 
CHLOROFORM AND ETHER. 


No. of Date. | Tissue. Solution. | Residue. 
extract. | 
1 25.1.16 | Rat uterus Lo Increase of tone. 
15.2.16 | "i | ee P. es 
16.2.16 Ms Diminution of height of 
contraction 
| 
2 15.2.16 . nee Increase of tone. 
16.2.16 5 Diminution of height of 
contraction | 
3 6.2.16 | Rat uterus oe | 1% Increase of tone. 
(Soxhlet) 
4 6.3.16 re Shght contraction | 2 be 5 
5 6.4.16 | Rat uterus ‘ f Wako s -. 


These results can be summarised as follows :— 

(1) Alcohol extract causes as a general rule either diminution or 
complete inhibition of the uterine contraction. Sometimes, however, its 
action is negative; in rare cases it produces increase of tone. 

(2) Locke solution of the extract of the residue of corpus luteum after 
extraction with alcohol always causes an increase of contraction of the uterus. 

In another series of experiments, extractions were made with alcohol 
to which a certain amount of water was added, the extractions being 
begun with absolute alcohol (purchased), the percentage of alcohol being 
gradually reduced. In this series the same sample of alcohol was used. 
The following results were obtained :— 

(1) 12.12.15. With absolute alcohol extract. No marked change; if 
anything, contraction. 

(2) Extract with 98 per cent. alcohol + 2 per cent. water per volume. 
No marked change: rate of rhythm perhaps somewhat slowed. 

(3) Extract with 96 per cent. aleohol + 4 per cent. water. Inhibition 
produced. The residue dried and extracted with Locke gave slight 
contraction. 

Repetitions of these experiments made 81.12.15 and 31.38.16 gave no 
change with the alcohol extract, and a sight contraction with the residue. 
Why inhibition was produced on the first occasion with this extract of 
96 per cent. alcohol + 4 per cent. water, but not subsequently, I am 
unable to explain. 

(4) Fresh corpus luteum was extracted with absolute alcohol in such 
a proportion that the resulting extract was one containing about 90 per 
cent. aleohol. This extract was dried and extracted with Locke solution. 
Both it and the residue after alcohol extraction produced an increase of 
tone when its action upon the rat’s uterus was investigated (31.10.15). 


Influence of Corpus luteum Extracts upon Plain Muscle 23 


CHLOROFORM EXTRACT. 


The extraction with chloroform was made as with alcohol, being 
frequently repeated either with cold chloroform or with the Soxhlet 
apparatus. The chloroform extract, after being dried and extracted with 
Locke solution, gave in no case any action on the uterus. The residue, 
after extraction with chloroform, dried and extracted with Locke, gave 
in four cases a distinct increase of tone, but in one case no effect was 


produced. 
ETHER EXTRACT. 

Ether extract was made in the same way as with chloroform, either 
by repeated extraction with cold ether or by the aid of the Soxhlet 
apparatus. Six experiments were performed with ether extract. The part 
of the dried extract which dissolved in Locke solution produced in three 
cases no change, in two a slight increase of tone, while in one the rhythmic 
contractions became somewhat slower. ‘The residue, after ether extraction 


and drying, extracted with Locke, gave in every instance a distinct 
increase of tone. 


WATER EXTRACT. 


The fresh or dried corpus luteum was either repeatedly extracted with 
cold distilled water or a decoction was made with boiling water. The solu- 
tions were filtered, evaporated to dryness over a water-bath, dissolved in 
Locke solution, and this again boiled and filtered. Seven experiments were 
performed with this water extract. In all but one an increased contraction 
was produced. In the exception, no action. The residue after extraction 
with ether was tested in one case; it produced no effect. The effects of 
water extract made in the above way were not so marked as those of 
similar extracts which had been evaporated to dryness in a vacuum at 
a temperature of 45° C. Two experiments were made with corpus 
luteum which, after extraction with chloroform, alcohol, and ether, and 
removal of proteins by lead acetate, was extracted with water. In both 
of these cases the rat’s uterus was caused to contract at a more rapid 
rate and with slight increase of tone. 


EFFECTS OF THE ASH. 


- Dried corpus luteum was incinerated in porcelain or silicate crucibles at 
a dull red heat. In one such estimation 3:578 per cent. ash was obtained. 
The ash was extracted with cold Locke solution, boiled and filtered. The 
effect of such solutions upon isolated portions of rat’s uterus was usually to 
produce inhibition: but only when the solution was strong, e.g. 5 per cent. 
of dry corpus luteum. This inhibition is probably a potassium effect, 
since analysis of the ash, for which I am indebted to Dr W. W. Taylor 
(see Appendix), shows a large preponderance of potassium. 


24 Itagaki 


SUMMARY. 


1. Extract of corpus luteum generally produces a distinct increase of 
tone in the surviving uterus of the rat, rabbit, cat, dog, and guinea-pig. 
Rarely, however, the opposite effect is produced. 

2. This difference of effect is not due to the condition of pregnancy 
or non-pregnancy, nor to varying strengths of the extract, but apparently 
to a difference in different samples of corpus luteum. It would appear, 
therefore, that there are two principles in the corpus luteum having an 
antagonistic action upon the contractions of the uterus. 

3. These principles can sometimes be separated by alcohol, the 
inhibitory material going into alcoholic solution. But this chalonic 
substance (17), which is soluble in water, is very small in amount. The 
hormonic substance, on the other hand, which is generally much larger 
in amount, is not soluble in alcohol nor in chloroform and ether, but 
is soluble in water. 

4. If we compare the action of corpus luteum extract upon the uterus 
with its effect upon other forms of plain muscular tissue, we find that it 
generally produces relaxation of the muscular tissue of the intestine and 
of the bladder of the rat, but contraction of the whole intestinal tube of 
the rabbit and kitten; although, when isolated strips of either the longi- 
tudinal or circular intestinal muscle of these animals were taken, they 
showed relaxation. Upon the iris of the frog no change could be observed. 

5. Injection of corpus luteum extract into a vein produces but little 
effect upon the blood-pressure ; if anything, there is a slight fall. 

6. A free secretion of milk is caused from the cut nipple of a lactating 
animal. 

7. Urinary secretion is not appreciably affected. 

8. I have been unable to obtain any definite proof that the action of 
corpus luteum upon the uterus is effected through the nerve-endings either 
of the sympathetic or of the pelvic nerve. 


The expenses of this investigation and of those described in the two 
succeeding papers have been assisted by grants from the Carnegie and 
Moray Research Funds. 


BIBLIOGRAPHY. 


(1) Prenant, Rev. gén. d. sci., 1898, t. ix. p. 646. 

(2) Stricken, Arch. f. Physiol., 1913, p. 259. 

(3) Fucus, Zeitschr. f. Geb. u. Gyn., 1914, Bd. Ixxv. p. 653. 

(4) GuaersBExe, ibid., p. 231. 

(5) Orr and Scorr, Monthly Cyclopedia and Med. Bull., 1911 and 1912, 
p. 207; Proc. Soc. Exper. Biol. and Med., 1911-12, vol. ix. p. 64; contrib. from 
Physiol. Lab., Med.-Chir, Coll., Philadelphia, 1912. 

(6) Magnus, Pfliiger’s Arch., 1904, Bd. cii. p. 123. 

(7) Kenrgr, Arch. f. Gyn., 1907, Bd. Ixxxi. p. 160. 


Influence of Corpus luteum Extracts upon Plain Muscle 25 


(8) Betow, Monatsschr. f. Geb. u. Gyn., 1912, Bd. xxxvi. p. 679. (Quoted 
from Biedl’s Innere Sekretion, 1916, ii.) 
(9) Brept, Innere Sekretion, 1916, ii. pp. 290, 292. 
(10) Sontckexe, Arch, f. Gyn., 1912, Bd. xevii, p. 409. 
(11) Orr and Scort, Proc. Soc. Exper. Biol. and Med., 1911-12, vol. ix. p. 63. 
(12) ScHArer and Mackenzig, Proc. Roy. Soc., 1911, B, vol. Ixxxiv. ; MACKENZIE, 
this Journal, 1911, iv. p. 305. 
(13) Laneciey and ANpErson, Journ. Physiol., 1895, vol. xix. p. 71. 
(14) James and Joun Gunn, Journ. Phar. and Exper. Therap., 1914, vol. v. p. 527. 
(15) VittEmin, Compt. rend. soc. biol., 1910, t. lxviii. p. 874. 
(16) Busquet, Biol. médicale, quoted from Villémin, 1910. 
(17) Scuirer, The endocrine organs, 1916, p. 5. 


APPENDIX. 
By W. W. Taytor, D.Sc. 


ANALYSIS OF ASH or Corpus LuTeuM or Ox. 
Gooch crucible . =13°7418 
+ substance. =13°9567 ‘2151 g. taken for analysis. 
(N.B.—Ash is hygroscopic, as original total 
weight was ‘2012 g.) 


Dry at 134°. . =13°9071 
1653 g. dry ash. 
0498 g. loss = moisture 23°15 per cent. 


Extract hot water =13°7455 
=e iy 


” 
*. ash almost completely soluble in water 


and quite so in dil. HCl. 
Added HCl to the extracts and evaporated to dryness in silica basin. 


Basin . : . =67-476) 
+ substance . =57°6427 ='1666 g. chlorides. 


Transferred to beaker, added acetic acid, ammonium oxalate. Filtered, washed, 
ignited precipitate. 
Platinum crucible =23°8552 
+ CaO =23°8560 0008 g. CaO = “35 per cent. Ca. 


Filtrate evaporated in silica basin, ignited with HCl. 
Basin . : . =97°4761 
+ chlorides . =57-6415 1654 ¢.=weight of NaCl+ KCl. 
Evaporated with sulphuric acid and ignited. 
+ sulphates . =57°6712 ='1951 g.=weight of Na,SO,+ K,SO, 
=23°6 per cent. NaCl+76'4 per cent. KCl. 
.*. the ‘1666 g. of mixed chlorides contained 


0016 g. CaCl, = 
0389 g, NaCl = 
1261 g. KC] = 
"1666 


‘35 per cent. Ca} 
9-2 oe Na 
39°7 K 


” 


ON THE ACTION OF VARIOUS EXTRACTS OBTAINED FROM 
THE COW’S OVARIES UPON THE MUSCULAR TISSUE 
OF THE UTERUS, INTESTINE, AND BLOOD-VESSELS. By 
M. Iracaki. (From the Laboratory of Physiology, Edinburgh 
University.) (With twelve figures in the text.) 


(Received for publication 28th July 1916.) 


IN connexion with my investigation on the action of corpus luteum 
extract (1) I had occasion to examine the influence of extracts from 
different parts of the ovaries upon the plain muscular tissue of the uterus, 
and incidentally of the intestine and blood-vessels. Since the action of 
certain of these extracts has not previously been investigated, it may be 
useful to give the results which I have obtained. The experimental 
methods were the same as those employed in the investigation of corpus 
luteum. 


EXTRACT OF HILUM OVARII. 


The part of the ovary adjacent to the hilum was cut out, rinsed with 
Locke solution or distilled water to free it from adherent fluid, lightly 
pressed with filter paper to get rid of superfluous water, weighed, 
minced, mixed with a definite volume of Locke solution, and boiled. After 
boiling, the extract was filtered and the filtrate made up to the original 
volume of the fluid by the addition of distilled water. 


Action on Uterus of Rat. 


A typical effect is shown in fig. 1, which shows an arrest of the 
rhythmic movements with relaxation of tone. In some cases there was 
only a diminution in the rate of rhythm and height of contraction, the 
relaxation of tone being absent. Occasionally the inhibition was followed 
by contractions, which gradually increased in extent (fig. 2). In all my 
experiments, only one case occurred in which there was an increase of tone 
from the beginning. In the pregnant uterus (one case) inhibition was 
also obtained. The strength of the extract used varied from 2 per cent. to 
10 per cent. 

It is a striking fact that, whilst extract of corpus luteum usually causes 
a remarkable increase of tone and sometimes of rate of rhythm, extract of 


28 Itagaki 


hilum shows a diminution in the rhythm of contraction, which may 
be accompanied by a marked relaxation of tone. Out of forty experiments, 
thirty-one showed diminution or inhibition of rhythmic movements, with 


Fic, 1.—Effect on cornu of rat-uterus of addition of extract of hilum ovarii to the Locke’s solu- 
tion in which the tissue was immersed. Notice the immediate cessation of the rhythmic 
movements and marked diminution of tone of the muscle. At L the extract was replaced 
by simple Locke’s solution. 

The time in this and in all the other experiments with the isolated tissue is marked 
in minutes. 


f\ 


a WAY Hl | 


Fic. 2.—Effect of extract of hilum ovarii (10 per cent.) upon surviving uterus of rat. The extract 
was applied at the first arrow (left) and replaced by simple Locke’s solution at the second. 
Notice that the rhythmic movements recommence even during the continuance of the extract. 


or without relaxation of tone; five showed inhibition of rhythmic 
movements followed after a time by a recurrence of contractions; in three 
no change was observed; in one, as above mentioned, there was increase of 
tone from the first. 


Action of Extracts of Ovary on Plain Muscle 29 


Action on Uterus of Rabbit. 


I have made tive experiments on the uterus of the rabbit, the strength 
of the extract varying from 5 per cent. to 10 per cent In four of these 


Fic. 3.—Etfect of extract of. hilum ovarii on uterus of rabbit, Notice the increase 
of tone. The extract was added at the first arrow and replaced by simple Locke 
at the second. 


a distinct increase of tone was produced by extract of hilum (fig. 3); in 
the fifth no distinct change. 


Action on Uterus of Cat. 


I have made two experiments on the excised cat’s uterus, the strength 
of the extract being 5 per cent. and 10 per cent. In both cases great 
increase of tone was produced by it. In addition to these two experiments, 
four others were made upon the uterus in situ (one of the cornua being 
connected by a thread with a muscle lever), an injection of 5 c.c. of a 
5 per cent. extract being made slowly into the jugular vein. In two of 
these experiments the uterine tone was increased. In the other two 
relaxation was produced, but in one of these the relaxation was preceded 
by an initial contraction (fig. 4). Two experiments were made with 
excised uteri of kittens. Here also an increase of tone was produced, 
although not nearly as well marked as with the uterus of the cat. 


Action on Uterus of Guinea-pig. 


A 5 per cent. to 10 per cent. extract was again used. In four 
experiments upon the uterus of the guinea-pig, all showed an increase of 


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Action of Extracts of Ovary on Plain Muscle 31 


tone. One of these experiments was made with a uterus which had been 
kept forty-eight hours in an ice-chest. 


Action on the Small Intestine. 
The experiments upon the small intestine were made partly with 
portions of the whole tube, partly with strips of the longitudinal coat. Of 
the experiments on the whole tube, three were performed with rabbit's 


Fic. 5.—Effect of extract of hilum ovarii on the intestine (whole tube) of the rabbit. Notice the 
increase of tone with gradual diminution of amplitude of the rhythmic movements. The 
first mark on the lowest line indicates the application of hilum extract, the second its 
replacement by simple Locke. 


intestine and two with the intestine of a kitten. All these, without 
exception, showed an increase of tone (fig. 5). The strength of the extract 
employed was from 3 per cent. to 10 per cent. Eight experiments were 
performed with longitudinal strips of the intestine of the rabbit, the 
strength of the extract used being from 2 per cent. to 10 per cent. All 
showed relaxation of tone, the “pendulum movements” being arrested 
(fig. 6) by the stronger solutions (above 5 per cent.), but not by the 
weaker (from 2 per cent. to 5 per cent.). 


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Itagaki 


Effects on Blood-pressure. 

Lambert (2) states, with regard to extract of ovary containing no 
corpus luteum, that this has no toxic and no physiological activity. 
Below (3), on the other hand, found that extract of ovary minus corpus 
luteum (“Proovar” as he terms it) causes a rise in blood-pressure and 
accelerates the pulse-rate. Bied1 (4), however, could not confirm this with 


Nh hi ane 


f t 


Fic. 6.—Effect of extract of hilum on a longitudinal strip of small intestine of 
rabbit. Notice the marked decrease of tone and cessation of rhythmic move- 
ments. At the second arrow the extract was replaced by Locke. 


an extract furnished to him by Poehl. I have performed in all seven 
experiments upon cats aneesthetised with chloroform, an injection of 
5-10 cc. of a 5 per cent. to 10 per cent. extract being made into the 
jugular vein. All show a distinct fall of blood-pressure (fig. 4), although 
the rate of the pulse is not diminished. The fall of blood-pressure was 
also produced after atropin had been previously injected. It is therefore 
a vasodilator effect. 


EXTRACT OF Liquor FOLLICULI. 


Extract of liquor folliculi was made in the following way. To a 
certain number of cubic centimetres of the filtered liquor obtained from 
large Graafian follicles Locke solution was added to make the fluid up to 
100 cc. This was then boiled and filtered, and the filtrate made up to the 
original volume by the addition of distilled water. 


Action on Uterus of Rat. 


I have made sixteen experiments upon the rat’s uterus, with extracts 
of from 2 per cent. to 12 per cent. of liquor folliculi. In most eases this 
extract produced an increase of tone in the uterus (fig. 7); when the 
rhythmic contractions were in abeyance, it had the effect of inducing 
these to reeommence. In one case, however, relaxation of the uterus was 
produced instead of contraction. The increase of tone was also obtained 
from a uterus in the pregnant condition. In two out of the sixteen cases 
no change was observed. 


Action of Extracts of Ovary on Plain Muscle 33 


Action on Uterus of Rabbit. 


The strength of the extract employed was from 5 per cent. to 


6 per cent. There were altogether five experiments. In four cases an 


Fic. 7.—Effect of liquor folliculi of cow on rat’s uterus suspended in Locke’s solution. 
Notice the increase of tone with cessation of rhythmic movements. 
The signal marks should be shifted a little to the left. 


Fic. 8.—Effect of liquor folliculi of cow (2 per cent.) on rabbit’s intestine, showing production of 
increase of tone, At the second mark the liquor folliculi was replaced by simple Locke. 


increase of tone as well as an increase in the height of the rhythmic 
contractions was observed. In the fifth case, on the other hand, inhibition 
was produced. 

Action on the Intestine. 


Four experiments were made on portions of the intestine: three from the 
rabbit, one from a kitten. All the tracings show distinct increase of tone 


(fig.8). The strength of extract employed was from 1:5 per cent. to 5 per cent. 
MOE: XI.) NO. 1,—1917. 


34+ Itagaki 


Action on Blood-pressure. 


Injection of from 5-7 ¢.c. of a 5 per cent. to 15 per cent. extract causes 
a marked fall of blood-pressure in the cat (fig. 9). 


me ith | 


Fic. 9,—Effect on blood-pressure of cat of an intravenous injection of 7 c.c. ofa mixture of 15 parts 
liquor folliculi and 85 of Locke. 


a, blood-pressure tracing ; 6, time in 10-sec. intervals; ¢, signal. 


ExTRACTS OF WHOLE OVARY. 


For this purpose, small ovaries were selected containing neither large 
corpora lutea nor large Graafian follicles. After being weighed, they 
were minced, boiled with Locke solution, filtered, and the filtrate made 
up to the original volume by the addition of distilled water. 


Action on Uterus of Rat. 


In four experiments this extract in every case produced diminution in 
extent or complete inhibition of the rhythmic movements (fig. 10). The 
strength of the extracts employed varied from 5 per cent. to 10 per cent. 


(ction of Extracts of Ovary on Plain Muscle 35 


Action on Uterus of Rabbit. 
Ott (5) noticed an increase of tone in the uterus (rabbit and eat he 
used an ovarian preparation made by Armour & Co. Bel! and Hick (6) 


failed to observe any etiect on the norma! uterus of the raodopit with an 


ie 
Tanne 
ti I Hie i 


Ki 


lt 


an 


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Fic. 11.—Effect of extract of whole ovary of cow on rabbit intestine (whole tube), At the first 
arrow a 10 per cent. extract was run in; at the second arrow it was replaced by Locke, 


ovarian preparation obtained from the Wellcome Laboratory, but in the 
pregnant and puerperal uterus of the same animal they found contrac- 
tion to be produced. Stickel (7) failed to obtain any effect on uterine 
movement in the rabbit by intravenous injection of an ovarian extract 
furnished by Hoftman, La Roche & Co. Fuchs (8) noticed inhibition of 


36 Itagaki 


the uterus in the rabbit when ovarian extracts obtained from Merck and 
Knoll were employed. 


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Out of five experiments made by me with extracts of strength varying 
from 5 per cent. to 10 per cent., four showed either an increase of tone or 


an increase of height of the rhythmic contractions. In one only there 
was no distinct effect. 


Action of Extracts of Ovary on Plain Muscle 37 


Action on the Intestine. 


I have made four experiments with the rabbit’s intestine, using the 
whole tube, and extracts of from 5 per cent. to 10 per cent. In every case 
an increase of tone was produced (tig. 11). 


Action on Blood-pressure. 


Gizelt (9) noticed a fall of blood-pressure to be produced by injecting 
ovarian extract into the dog. Schickele (10) found that extract of cow’s 
ovary made with cold physiological salt solution produced only a slight and 
evanescent fall of blood-pressure; sometimes there was a rise instead of a 
fall. On the other hand, the expressed juice of the ovary obtained by the 
use of high pressure caused a fall of blood-pressure which continued for 
several minutes, even when a minute dose was given. Schickele was only 
able to obtain this result in the dog and rabbit, and Bied] (11) considers 
the effect to be non-specific. Im my own experiments, an intravenous 
injection of 5 c.c. of a 5 per cent. to 10 per cent. extract always produced 
a marked fall of blood-pressure in the cat (fig. 12). 


SUMMARY. 


1. Extract of hilum ovarii has the opposite effect to extract of corpus 
luteum upon the movements of the rat’s uterus, for it causes inhibition 
instead of contraction. The uterus of other animals is attected differently, 
for in the rabbit, cat, and guinea-pig an extract of the hilum usually has 
the result of producing an increase of tone. 

2. Liquor folliculi produces an increase of tone of the uterine muscle in 
the rat, rabbit, and cat, or at any rate an increase in height of the rhythmic 
contractions. 

3. When applied to the whole thickness of the intestinal tube, all the 
extracts tested—hilum, liquor folliculi, and whole ovary—determine an 
increase of tone, whereas a strip of the longitudinal coat of the rabbit's 
intestine is sent into relaxation by extracts of hilum. 

4. A fall of blood-pressure is produced by intravenous injection of all 
the extracts employed—hilum, liquor folliculi, and whole ovary. 


BIBLIOGRAPHY. 


(1) Iraeak, this Journal, preceding paper. 

(2) Lampert, Compt. rend. soc. biol., 1907, Ixii. 18. 

(3) Betow, Monatsschr. f. Geb. u. Gyn., 1910, xxxvi. Quoted from Bied1], 
Innere Sekretion, 1916, ii. 290. 


38 


Action of Extracts of Ovary on Plain Muscle 


(4) Briepn, loc. cit. 

(5) Orr, Journ. Exper. Med., 1909, ii. 326. 

(6) Betu and Hick, Brit. Med. Journ., 1909, 1. 777. 

(7) Sticket, Arch. f. Physiol., 1913, p. 259. 

(8) Fucus, Zeitschr. f. Geb., 1914, Ixxv. 653, 

(9) Grizeut, Pfliiger’s Archiv, 1913, cli, 562. 
(10) ScuickeLe, Miinch. med, Wochenschr., 1911, No. 3. 
(11) Brept, Innere Sekretion, 1916, ii. 292. 


THE ACTION OF CERTAIN GLAND EXTRACTS AND DRUGS 
UPON THE UTERUS OF THE RAT. By M. Iracakt. (From 
the Laboratory of Physiology, Edinburgh University.) (With four 
figures in the text.) 


(Received for publication 31st July 1916.) " 


THE uterus of the rat has been used for the investigation of drugs by 
Callibureés (1), Franz (2), Dale (3), Fiihner (4), Guggenheim (5), 
Gunn (6), Herring (7), although most observers have employed the uterus 
of other animals for this purpose. In my experiments on the corpus 
luteum (8) and extracts of ovary (9) I chiefly made use of the uterus of the 
rat, and in connexion with those experiments had occasion to make a 
certain number of observations upon the effect of other animal extracts 
and drugs upon it; the results of these are given here. The technique 
employed was the same as in the other experiments. 

The uterus of the rat offers certain advantages over that of larger 
animals. (1) It can generally be easily obtained. (2) Being small, only 
a comparatively small quantity of the solution to be tested is required. 
(3) The spontaneous rhythmic contractions which it shows in oxygenated 
Locke solution at 37° C. are from the beginning of immersion fairly regular, 
whereas, as Kehrer has pointed out, those of the uterus of the dog and 
rabbit are apt to be irregular (10). Any change, therefore, which is caused 
in the uterine movements can be easily observed. 

Fig. 1 is a typical tracing exhibiting the movements of the rat’s uterus 
under the above circumstances. It will be seen that it shows quite regular 
rhythmic contractions. The ascending part of each curve indicates con- 
traction; the descending part, relaxation. The contraction starts fairly 
quickly, slows down towards the apex of the curve, and, after the 
maximal contraction is reached, comes down at first rapidly and then 
more gradually. With the weight of lever employed by me the duration 
of contraction was shorter than that of relaxation. The rhythm of the 
contractions of the rat’s uterus under these circumstances is, roughly, one 
per minute. 

Incidentally, I find that if the uterus is excised and kept in Locke 
solution in an ice-chest at a temperature of between 3° and 7° C., even for 
as long as three days, it still responds when immersed in oxygenated Locke 
solution at 37°C. A somewhat similar observation was made by Hudston, 


40 Itagaki 


who was able to demonstrate the action of drugs on an excised uterus 
(guinea-pig) which had been kept as long as seven days in an ice-chest (11). 


Fic. 1.—A typical tracing of the movements of a piece of cornu of surviving rat-uterus, immersed 
in oxygenated Locke’s solution at a temperature of 37° C. The movements are amplified 
about four times. Time in minutes, The ascending part of each curve indicates contraction. 


Kurdinowsky (12) also found that the uterus of the rabbit kept for from 
twenty-four to forty-eight hours in the cold still showed spontaneous 
movements. 


EXTRACTS OF THE POSTERIOR PART OF THE PITUITARY Bopy. 


A large number of observers have corroborated the original observation 
of Dale that intravenous injection of extract of the posterior lobe of the 
pituitary body causes marked uterine contraction; a similar result being 
obtained with a portion of the uterus immersed in the extract. Most of 
these experiments have been made in the cat, rabbit, and guinea-pig; but 
Dale, Guggenheim, Herring, and Gunn have seen the same thing in 
the rat. 


These observations I can entirely corroborate. I have employed 
extracts of the dried posterior lobe of from 0:1 to 1 per cent. strength, 
and extract of fresh ox pituitary of 3 per cent. strength. The extracts 
were made by boiling with Locke solution and afterwards filtering. In 
most cases not only is the tone immediately increased when the portion of 
uterine cornu is immersed in the extract, but the muscular tissue usually 


Action of certain Gland Extracts and Drugs upon Uterus of Rat 41 
goes into persistent contraction : this howe ver presently olivVeS WAV to a 
rhythmic contraction, at first of small amplitude, but gradually increasing 


in extent (fig. 2). Sometimes the increase of tone is maintained for a 


Fic. 2.—Effect on cornu of rat-uterus suspended in Locke’s solution of the addition of 
extract of posterior lobe of pituitary of ox. Time in minutes. 


considerable time. The contraction caused by the extract of pituitary 
is somewhat similar to that of extract of corpus luteum. 

Extract of the anterior lobe of the pituitary body does not, so far as 
my observations go, produce any eftect. 


ADRENALIN. 


A large number of experiments have been made by previous workers 
regarding the action of adrenalin upon the uterus. Most have been upon 
the cat, rabbit, and guinea-pig, but some upon the human uterus, and one 
series upon the rat’s uterus. ‘The general results can be summed up in 
the following table, which is based on one given by Gunn (6) :— 


Results. 
Name of animal. 
Non-pregnant. Pregnant. 

Rabbit | Contraction | Contraction. 
Cat . ; ; ; Inhibition - 
Guinea-pig vi a Inhibition. 
Rat . = : i ee _ r 
Human . , 3 Contraction 
Ferret . : =| a 
Dog. : : < 


Monkey . 9 


42 Itagaki 


In my experiments on the uterus of the rat, Parke, Davis & Co.’s adrenalin 
has been employed, the strength of the solution used being from 1 : 100,000 
to 1:10,000. The result in this animal is always to produce inhibition 
of the rhythmic movements (tig. 3), with or without relaxation of the 


——— a rt 
Adromolin /:.50000 


Fic, 8.—Effect of adrenalin (1 in 50,000) on suspended cornu uteri of rat. Notice the 
complete inhibition of the rhythmic movements. 
general tone of the muscle, and this whether the uterus were from a 
pregnant animal or not. As we have seen (8), extract of corpus luteum 
produces an increase of tone in nearly all cases in the rat, so that 
in this respect adrenalin has an effect directly contrary to that of corpus 
luteum extract. 
THYROID EXTRACT. 

Lindemann and Aschner (13) state that extract of thyroid, freed 
from peptone and protein, brings on labour pains in the parturient subject. 
Bell and Hick (14), who employed preparations obtained from the Well- 
come Laboratory, failed to obtain any result upon the uterus of the rabbit 
in the non-pregnant condition, whereas increase of tone was obtained in uter1 
taken from pregnant and puerperal animals. Guggisberg (15) describes an 
increase of tone in the uterus of the guinea-pig as the result of immersion 
in expressed juice of sheep thyroid diluted with physiological salt solution. 
Mosbacher (16), using the thyreoglandol preparation of Hoffmann, La 
Roche & Co., was unable to obtain any effect as the result of intravenous 
injection in rabbits, but sometimes got contraction in the excised uterus 
immersed in the solution. Fuchs (17) also describes contraction of the 
uterus of the rabbit as being obtained on immersion of the excised organ 
in expressed thyroid juice. 


In my own experiments fresh sheep’s thyroids were minced, boiled with 
Locke solution, and filtered. With extracts of such strengths as 0°4 per 


Action of certain Gland Extracts and Drugs upon Uterus of Rat 43 


cent. to 0°8 per cent. no effect was perceptible. Even with strengths of 
from 1 per cent. to 5 per cent. there was generally very little effect pro- 
duced, although occasionally a tone increase was observable at the beginning 
of immersion, but this was soon recovered from. ‘The results with boiled 
thyroid extract must therefore be considered negative. 


EXTRACT OF ORCHITIC SUBSTANCE. 


For the investigation of the action of this substance I have used desic- 
cated testicle-substance of lamb (Armour & Co.). Extracts were made by 
boiling with Locke solution and subsequently filtering. Even with so 
strong a dose as would correspond to 10 per cent. fresh gland I have failed 
to get any noteworthy effect on the uterus of the rat, although Ott (18) 
states that extracts of this substance produce a slight increase of tone in 
the uterus of the rabbit and cat. 


Extract OF UTERUS. 

Guggisberg (15) found uterine extract from the cow to have no action 
upon the excised uterus of the guinea-pig, but in one experiment in which 
an extract was made from the uterus of a gravid cow an increase of tone 
was observable. 


My own experiments were made with extracts of the uterus of the rat. 
One of the cornua was minced, boiled with Locke solution, and the extract 
filtered or decanted and kept at 37°C. This extract was passed into Locke 
solution in which the other uterine cornu of the same animal was immersed. 
The strength of the uterus extract was about 0-2 per cent. Out of five 
experiments, in four no effect was observable; in one relaxation of the 
general tone of the muscle was produced, the rhythmic movements being 
maintained. 


EXTRACT OF CEREBRAL SUBSTANCE. 
Ott (18) states that he was able to observe a marked increase of con- 


traction in the excised uterus of the cat and rabbit on immersion in extract 
of dried brain-substance (Armour & Co.). 


For my own experiments a Locke extract of tabloid cerebrin of sheep 
(Burroughs, Wellcome & Co.) was employed, the extract being as usual 
boiled and filtered. This had practically no influence upon the rat’s 
uterus. 

NICOTINE. 

Franz (2) noticed that in the uterus of the rabbit intravenous injection 
of nicotine produced powerful contractions. He obtained the same result 
with portions of excised uterus both rabbit and human. Cushny (19), 
also using the method of injection, found that in the cat both nicotine and 
adrenalin act upon the uterus in the same way as pilocarpine, their effects 


Ad, Itagaki 


agreeing generally with that of stimulation of the hypogastric. Kehrer 
(10) states that nicotine first causes inhibition, afterwards contraction, of 
the excised uterus of the non-gravid cat. Sugimoto (20) found the action 
of nicotine to be very slight on the excised uterus of the guinea-pig, 
although intravenous injection caused distinct contraction. Gottlieb and 
Meyer (21) state that in the uterus of the non-pregnant cat nicotine first 
produces inhibition and then contraction, whereas its effect upon the gravid 
uterus is to cause immediate contraction. 


My own experiments show that in the rat solutions of a strength vary- 
ing from 1:10,000 to 1:5000 cause an increase of tone. With stronger 
solutions, such as 1: 1000, there is at first an arrest of contraction and then 
an increase of tone, but with very strong solutions (e.g. 1: 200) inhibition 
alone is produced. 

PILOCARPINE. 


Brennecke (22) and Kleinwachter (23) state that labour pains are 
strengthened by intravenous injection of pilocarpine. Dale and Laidlaw 
(24) and Kehrer (10) and Fardon (25) investigated the influence of 
pilocarpine upon the isolated uterus: they found that in the cat the con- 
tractions are increased, or if in abeyance are excited. Dale and Laidlaw 
obtained the same effect with the uterus of the guinea-pig, and this whether 
the drug were injected into a vein or whether the excised uterus were 
immersed in the solution. Gunn (6) got little effect upon the rat’s uterus. 


In my own experiments the results of pilocarpine have been inconstant. 
I have used strengths of solution from 1: 100,000 to 1: 1000 made up with 
Locke. The result has generally been to produce contraction, but occasion- 
ally to diminish it. 

ATROPINE. 

Rohrig (26) states that intravenous injection of 0:003 grm. of atropine 
sulphate causes cessation of the peristaltic movements of the uterus of the 
rabbit. Direct application seemed to have no effect. Franz (2), who also 
used the uterus of the rabbit, obtained a different result from intravenous 
injection of atropine, which he describes as having no effect. Kehrer (10) 
states that a weak solution (1: 250,000) causes contraction; a medium 
strength (from 1:10,000 to 1:25,000), an increase of tone; but even a 
solution of much greater strength (such as one over 1:2500) does not 
arrest the contractions (in the cat and dog). Kurdinowsky (27), using 
rabbit’s uterus, found the action of atropine sulphate in doses of from 
0:005-0:08 grm., when intravenously injected, to be much the same as that 
of morphine. Quagliariello (28) found the action of atropine on the uterus 
uncertain, although when in a condition of increased tone there was some- 
times a tendency to produce relaxation. 


In my own experiments upon the uterus of the rat I have used 
strengths of atropine sulphate of from 1: 20,000 to 1:1000. With a 


Action of certain Gland Extracts and Drugs upon Uterus of Rat 45 


strength of 1:20,000 the individual contractions are increased in height 
without the cveneral tone being atfected. With a strength of 1 10.000 
there is a slight diminution in the height of the contractions; but even with 
a strong solution, such as 1 : 1000, I have failed to get complete inhibition. 


BARIUM CHLORIDE. 


Kehrer (10) describes the production of strong contraction in the 
uterus of the cat by solutions of this salt. 


The strength of the solutions which I have used for the excised rat- 
uterus has been from 1:100,000 to 1:1000. Even with a strength of 


Fic. 4.—Effect of barium chloride (1 in 20,000) on the excised uterus of rat. The drug was added 
to the Locke’s solution at the first mark and removed at the second. Notice not only an 
increase of tone of the muscular tissue, but also an increase of rate of the rhythmic contractions. 

1:100,000 an increase of tone is obtained; with stronger solutions, the 

uterus passes into a condition of pronounced contraction (fig. 4). 


SUMMARY. 

The results of these experiments upon the rat-uterus may be briefly 
stated thus :— 

1. Extract of posterior lobe of pituitary always produces an increase 
of tone. 

2. Solutions of adrenalin inhibit the rhythmic contractions (with or 
without diminution of tone). 

3. Extract of thyroid occasionally produces an increase of tone, but 
generally has no effect. 


46 Action of certain Gland Extracts and Drugs upon Uterus of Rat 


4, Extracts of orchitic substance, of uterus, and of brain have no 
appreciable effect. 

5. The action of nicotine is variable. Weak solutions produce an 
increase of tone. Stronger solutions cause first inhibition, then increased 
contraction. Still stronger solutions produce inhibition alone. 

6. The action of pilocarpine is inconstant, but usually an increase of 
tone is produced. 

7. Weak solutions of atropine produce increase of the rhythmic con- 
tractions; stronger solutions, a slight diminution; but even with a very 
strong solution there is no absolute arrest of the contractions. 

8. Barium chloride in all strengths which produce any effect causes 
increased tone and stimulates the rhythmic contractions. 


BIBLIOGRAPHY. 


(1) Catiisurczs, Compt. rend. Acad. d. Sci., 1857, xlv. 
(2) Franz, Zeitschr. f. Geb., 1904, lil. 361. 
(3) Dats, Biochem. Journ., 1909, iv. 427. 
(4) Fiuner, Therap. Monatssch., 1913, xxvii. 202. 
(5) GuecenHeiM, ibid., 1912, xxvi. 795. 
(6) Gunn, Journ. Pharm. and Exper. Therap., 1914, v. 527. 
(7) Herring, this Journal, 1914, viii. 267. 
(8) IvaGakl, this Journal, 1917, xi. 1. 
(9) Iracakt, ibid., 27. 
(10) Kexrer, Arch, f. Gyn., 1907, Ixxxi. 160. 
(11) Hunsroy, from Gunn, Proc. Roy. Soc., 1914, lxxxvil. 571. 
(12) Kurprnowsky, Zentralbl. f. Physiol., 1904, xviii. 3. 
(18) LinpeMann u. ASCHNER, Miinch. med. Wochenschr., 1913, 1x. 2979. 
(14) Buair Bett and Hick, Brit. Med. Journ., 1909, i. 777. 
(15) GueeisBere, Zeitschr. f. Geb., 1914, Ixxv. 231. 
(16) Mospacuer, ibid., p. 362, 
(17) Fucus, ibid., p. 653, 
(18) Orr, Journ. Exper. Med., 1909, xi. 326. 
(19) Cusuny, Journ. Physiol., 1910, xh, 237. 
(20) Sueimoro, Arch. f, exper. Path, u. Pharm., 1913, Ixxiii. 27. 
(21) Gorriies u. Meygr, Pharmakologie, 1910, p. 186. 
(22) Brennecxg, berl. klin. Wochenschr., 1880, p. 122. 
(23) Kierswacuter, Arch. f, Gyn., 1878, xiii. 280. 
(24) Dave and Larpiaw, Journ. Physiol., 1912, xlv. 1. 
(25) Farpon, Biochem. Journ., 1908, iii. 405. 
(26) Rouric, Virchow’s Arch., 1879, Ixxvi. 1. 
(27) Kurpinowsky, Arch. f. Gyn., 1906, Ixxx. 289. 
(28) QuaGLiaRIELLo, Arch, di Ostetricia e Ginecologia, 1916, v. 3. 


THE EFFECT OF THYROID-FEEDING ON THE WEIGHT OF 
THE SUPRARENALS AND ON THEIR ADRENALIN CON- 
TENT, By P. T. HERRING. (From the Physiology Department, 
University of St Andrews.) 


(Received for publication 18th August 1916.) 


InN a previous paper the author (11) showed that the administration of 
raw thyroid in large doses to cats increases the amount of adrenalin in 
the suprarenals of these animals. There was also evidence that it 
increases the weight of the suprarenals. The importance of these 
conclusions led the author to investigate the effect of smaller doses of 
thyroid in a more extensive series of experiments. The results upon the 
suprarenals are recorded in this paper. 

That administration of thyroid causes hypertrophy of the suprarenals 
in new-born animals was shown by Hoskins (13) in 1910. Hoskins 
fed guinea-pigs from the day of birth for 15 days with small amounts, 
5 to 15 mg., of desiccated thyroid, and then weighed the suprarenals, using 
as controls guinea-pigs of the same age. He obtained an average 
hypertrophy of 25 per cent. in the suprarenals of the thyroid-fed animals. 
Further experiments showed that adult guinea-pigs fed with thyroid 
throughout pregnancy give birth to young in which the average weight 
of the suprarenals is below the normal. This deficiency in weight was 
interpreted by Hoskins as indicating a reaction in the suprarenals of the 
offspring to increase of adrenalin in the blood of the mother brought 
about by thyroidism. Hoskins recorded other experiments which were 
inconclusive, but believed that his evidence on the whole supported the 
theory that the thyroids normally stimulate the suprarenals to hyperplasia, 
though he made the reservation that the hypertrophy might be due to 
toxin in the thyroid employed. 

A close relationship between the functions of the thyroid and 
the medulla of the suprarenals is suggested by the work of Asher and 
Flack (1). Several observers, Fraenkel (9), Broking and Trendelen- 
burg (2), and Krause (14), employing different methods, have recorded 
an increase of adrenalin in the blood of patients suffering from 
exophthalmic goitre, a disease which most authorities believe to be 
characterised by excessive thyroid secretion. Ott and Scott (15) further 
ascertained that intravenous injection of thyroid extract increases the 
adrenalin in the blood of experimental animals; but they also obtained a 
similar increase by injecting extracts of other organs. Sir Edward 


48 Herring 


Schafer (16), in summing up the evidence of the action of the thyroid 
upon the suprarenals, states that “it may be assumed that the secretion 
of the thyroid in exophthalmic goitre acts as a direct stimulant to the 
suprarenal capsules, causing them to yield adrenalin to the blood in larger 
quantity ; a result which is also obtained by thyroid feeding.” 

That the suprarenals readily give off adrenalin into the blood has been 
frequently shown. Elliott (7) found such to occur as the result of the 
administration of anesthetics. Cannon and de la Paz (3) detected a 
rise of adrenalin in the blood following upon emotional influences. 
Cannon and Hoskins (4) showed an increase in the blood of cats as the 
result of asphyxia and stimulation of the sciatic nerve. The amount of 
adrenalin in the blood is therefore lable to vary. An increase in the 
blood does not necessarily indicate an increase of adrenalin in the body. 
A better index of any change in the adrenalin content of the body is 
furnished by an estimation of the amount in the suprarenal capsules and 
chromaphil tissue. Moreover, measurements of adrenalin in the blood are 
not very satisfactory, partly because of the minute quantity present, and 
partly because of the methods employed. Schafer (op. cit.) utters the 
caution that most of the physiological tests which have been used for 
measuring adrenalin in the blood would also be given by pituitrine. 

In the following experiments the weights of the suprarenals of normal 
and thyroid-fed animals are recorded and compared. The adrenalin of 
the suprarenals is estimated by Folin’s micro-chemical method. 


PROCEDURE. 


The animals used are white rats. These were selected partly because 
of their small size and the convenience with which they can be observed 
and fed, but also because their response to small measured doses of thyroid 
has already been recorded by Hewitt (12). A further advantage in their 
use lies in the fact that many details, more especially as regards age, size, 
and weight of body and organs of the white rat, have been compiled by 
Donaldson (6) in the Memoirs of the Wistar Institute. The rats selected 
were all males. The female rat has relatively larger suprarenals than the 
male, the difference tending to increase as age advances (Donaldson). 
The animals reserved for thyroid-feeding were of various ages, but were 
mostly young adults. A like number of animals of as near the same ages 
as possible were kept as controls. Both sets were given a supply of bread 
and milk more than necessary for their daily requirements. The one set 
received each a daily addition of 0°5 grm. fresh ox thyroid finely minced 
and mixed with the bread and milk. The amount of thyroid was fixed 
at 0°5 grm. fresh gland because it is about the upper limit of the dose 
which may be given without producing loss of weight of the animal. 
The feeding was continued for from 20 to 30 days. Each thyroid-fed 
animal was kept in a separate metabolism cage so as to ensure that it 


Effect of Thyroid-Feeding on the Suprarenals 49 


received the amount of thyroid desired. The control animals were kept 
in several cages in the first experiment (Table I.), and no addition was 
made to their diet of bread and milk. 

In a second series of experiments, recorded in ‘Tables III. and IV., ten 
male rats taken from two litters born on the same day were utilised. 
Five of them, averaging 73 grm. each in body-weight, received 0:2 grm. 
fresh ox thyroid daily for from 23 to 26 days. The other five, which 
averaged the same weight, received 0°2 grm. fresh ox flesh daily for the 
same period. All these animals were kept under identical conditions. 
Each was in a separate metabolism cage and received the same diet of 
bread and milk, with which the thyroid was mixed in the one case, and 
the ox flesh in the other. 

At the close of the experiment the rat to be examined was decapitated, 
and the suprarenals dissected out, cleared of the surrounding fat, and 
weighed. The suprarenals were then minced and pounded in a mortar, 
extracted with decinormal HCl, boiled with the addition of 10 per cent. 
sodium acetate, cooled and filtered. The filtrate, with the addition of 
Folin’s reagent and saturated sodium carbonate, was made up to 50 c.c. 
with distilled water. The colour resulting was then compared in a 
Dubosq colorimeter with the colour produced by a solution of uric acid 
containing 0°5 mg. per cent. This is half the strength recommended by 
Folin (8), but for measuring the small amount of adrenalin in the rat's 
suprarenals is more suitable than the usual standard. The depth of the 
uric acid solution was set at 20 mm. in the colorimeter, and the depth of 
the suprarenal solution regulated until of the same tint. The amount of 
adrenalin in the solution was then determined, taking the colour produced 
by the adrenalin as being three times as intense as that given by the same 
strength of uric acid. E. Merck’s extra pure uric acid was used as the 
standard, and it is assumed in these experiments that what Folin found 
true of Kahlbaum’s pure uric acid is also true of E. Merck’s. The uric 
acid solution was prepared according to the directions given by Folin, 
and was never more than a day old when used. It deteriorates rapidly, 
and shows considerable loss of strength three days after being made up. 

All the quantitative measurements of adrenalin given in this paper 
have been made by Folin’s method. Several contro] experiments have 
been carried out in which the solutions have been tested by Elliott's 
method on the blood-pressure of a pithed cat. Parke, Davis & Co.'s pure 
adrenalin powder was used to make up the standard solution. It was 
diluted to a strength similar to- that found in the extract of suprarenal 
tested by Folin’s method, and both were injected intravenously. The 
action of the pure adrenalin solution on blood-pressure was rather greater 
than that of the extract of suprarenal. It cannot be said that the controls 
were quite satisfactory. This was chiefly because, in the absence of any 
assistance, the experiment took a considerable time. The adrenalin 


solutions were made up much later than the extract tested by Folin’s 
VOL. XI., NO. 1.—1917. 4 


50 Herring 


method, and the adrenalin in the latter may have perished to some extent. 
Folin found the method very reliable. The figures given in the following 
tables for the adrenalin content of the normal rat show remarkable con- 
sistency. For comparative estimations of adrenalin the method appears 
to be the most satisfactory yet devised. 


RESULTS OBTAINED. 

The results of the first series of experiments are recorded in Tables I. 
and II. Thirteen male rats were examined in each case. In weight the 
normal animals varied from 107 to 294 grm., while the thyroid-fed rats 
varied from 96 to 294 grm. final weight. No record is kept of any change 
in weight of the normal animals, but in the thyroid-fed rats the original 
body-weight is given in column 4, the final weight in column 5, and the 
difference in column 6. The total weight of the suprarenals in mg. is then 
given. The weight of the suprarenals in mg. per 100 grm. body-weight 
is recorded in the next column. The total amount of adrenalin in mg., 
the adrenalin in mg. per 100 grm. body-weight, and the percentage of 
adrenalin in the suprarenals are given in the last three columns re- 
spectively. 

Some of the thyroid-fed animals lost weight during the experiment, 
but the majority gained, although the average gain in the thirteen rats 
only works out at 3 grm. The last animal of the series was a much older 
animal than the rest, and was very fat when first taken. It lost 49 grm. 
in 21 days, but was quite healthy when killed. No. 7 rat, which gained 
42 grm., died suddenly on the 20th day of the experiment. Two other 
animals, while apparently thriving and in good health, died suddenly at an 
earlier stage and are not recorded in this experiment. The average weight 
of the normal rats in Table I. is 192 grm., that of the thyroid-fed rats 
171 grm., showing a difference of 21 grm. 

The second series of experiments is recorded in Tables III. and IV. In 
these the length of each animal, taken from tip of nose to anus in a 
straight line, is recorded in column 7. This measurement is an important 
one in Donaldson’s statistics, and is correlated in his tables with the 
age, weight of body, and weight of organs of the animal. The animals in 
this series were rather under three months old when killed, and agree 
fairly closely in age, length, and body-weight with the corresponding 
normal animals of Donaldson’s. The thyroid-fed animals received in 
their diet 0-2 grm. fresh ox thyroid daily. This amount seems to have 
retarded slightly the growth of some of them. The thyroid-fed animals 
gained an average of 62 grm. in body-weight in from 23 to 26 days, 
while the control animals, which received in their diet 0:2 erm. fresh 
ox flesh daily, gained an average of 70 grm. during the same _ period. 
Both sets started with the same average weight; the heaviest rat, 
No. 3 of the thyroid-fed ones, retained its position throughout the 


Effect of Thyroid-Feeding on the Suprarenals 51 


experiment. This animal died suddenly on the 24th day of the ex- 
periment ; its suprarenals were examined some hours later. It appeared 
the strongest animal of the series. All were apparently healthy, and 
only differed in appearance from the normal animals in the greater 
roughness of their coats. The thyroid-fed animals consumed much more 
food than the control rats, but were somewhat thinner. 


WEIGHT OF THE SUPRARENALS. 


In the normal animals of the first series the weight of the suprarenals 
varies from 21 to 41 mg., the average for the thirteen rats being 29 mg. 
According to Donaldson’s statistics the weight of the suprarenals of the 
male rat after 40 to 50 days of age increases fairly uniformly with the 
age and weight of the animal. Thus the average weight of the supra- 
renals of a rat with a body-weight of 107 grm. is 22 mg., which is the 
figure found in the first rat of the normal series. For a rat of 290 grm. 
weight the suprarenal average weight is 44 mg. It will be seen that the 
figures recorded in Table I. agree very closely with the averages given by 
Donaldson. 


TaBLE I.—Frirst Serres. NorMAL Rats FED ON BREAD AND MILK ONLY. 


1. 2. | 3. eo em eer: oa ee, Pei es 
é | Weight of (Adrenalin | Percentage 
Boy. Seen ok suprarenals Total | Bee Ors “deanalin . 
Number.| Sex. | weight in |." © | inmg. per | adrenalin! 100 grm. : 
suprarenals 100 mend Sea, to hh in 
ae in gr Oe Be OY suprarenals. 
in grin. | body-weight. | weight. ~ 
1 M. 107 22 20°56 0:036 0°033 0163 
2 M. 117 21 17:94 0°031 0°026 07147 
3 M. 128 23 17:96 0°043 0:033 0°187 
4 M. 129 29 22°48 | 0046 0°036 07158 
oe f. 4M. 148 22 14°86 0-049 0°033 0°222 
6 M. 153 29 18°95 | 0075 0°049 0°258 
ih M. 158 26 16°45 0075 | 0047 0°288 
8 M. 171 21 12°28 0046 § 0027 0219 | 
9 M. 239 28 ibs Leg E 0065 | 0027 | 0232 | 
10 M. 275 31 11°27 0092 | 0-033 0-297 | 
11 M. 285 40 | 14:03 | 0098 | 0034 | 0:245 
12 M. 287 41 14:28 GOyo. , OO? |. Game 
13 M. 294 39 13°26 | 07110 0037 | 0°282 
| | | | 
Averages : 192 29 15710 0°065 0034 _ 0224 


i 


In the second series of normal rats the weight of the suprarenals is 
very constant at about 25 mg., a figure rather less than Donaldson’s, 
which is 27 mg. for a rat with a body-weight of 143 grm. 

The effect on the weight of the suprarenals of feeding with small doses 
of thyroid is very distinct In the first series of animals (Table IL.) the 


52 Herring 


weight of the suprarenals varies from 37 to 60 mg., the average for the 
thirteen rats being 46 mg. as against 29 mg. in the normal. The difference 
is even more pronounced when the weights per 100 grm. body-weight 
are compared. In the normal animals the average weight of the supra- 
renals per 100 grm. body-weight is 15:1 mg., while in the thyroid-fed 
rats it is 26°88 mg. The aifference is 11°78 mg., giving an increase in the 
thyroid-fed rats of 78 per cent. 


TABLE ]].—Firstr SerIES. RATS FED ON BREAD AND MILK +0°5 GR. FRESH 
Ox THYROID DAILY. 


~sJI 
19.2) 
Ne) 
tear 
° | 
4 


6. 


Eee 3 4 ) 

| a= | a 9 ‘ f=] e's 

lf 1.12 |@ |ee. | Soe |e | ee ee 
5 | SS | 22) Pe lSe) Peni 2822] Be aes ee 
ret Oss) aH ae |e es aaa es SH | Segs | oH | ao SS 
A Pg | be | eo ee oe eo | 8 2738 | aoe 

| o He ey Oo ‘Si eS ° hol tet on 

} o | = = Sal o xq 5 
1 | M.} 830days| 96 96 0) 37 38°54 0:063 | 0:066 | 0170 | 
PM PO. | OS) 124 seo aaa 33°87 0:075 | 0:061 | 0-178 | 
SelM een est nl) 132. ise ual 3 28:78 0057 | 0:043 | 07150 | 
4) Mi 83= 50") 419) | 186") 17 5 41:91 0072 | 0:053 |) 0-126) 
BM 22s) dole |) 400 ai 52 37°14 0:088 | 0:063 | 0-169 | 
Gale 23) Ne AS8e bd ei ly 40 25°80 0:069 | 0:044 | 0-172 | 
| Mee 20) 4 | 1837) -p do 60 32°78 0128 | 0070) |/-0-2is 
SM oe 185 | 189 | + 4 39 20°63 0-061 0:032 | Os1565)) 
9|M.| 20 ,, | 220 | 190.| —30 38 20:00 0080 | 0-042 | 0210 | 
10) Mi.) 20° 5, | 155 | asl" | 4-36 47 24:60 07133 | 0:070 | 0-283 | 
1 eMs 23 | 204: 83 | — 11 42 21-76 0-092 | 0:048 | 0-219 
12 | M.| 21 ,, | 199 | 205 | + 6 52 25°36 0092 | 0:°045 | 0177 
13 | M.| 21 ,, | 343 | 294 | —49 5d 18°70 0138 | 0-047 |\ 0-251 
Averages . Sy GSan | aT |) es 46 26°88 0088 | 0:052 | 0191 


In the second series the same effect is noted. The suprarenals of 
the thyroid-fed animals vary from 32 to 60 mg. in weight, the average 
being 41 mg. compared with the average of 25 mg. in normal animals of 
the same age. Per 100 grm. body-weight the figure for the thyroid-fed 
animal is 80°38 mg. compared with 17-27 mg. in the normal of this series. 
This is an increase in weight of the suprarenals of the thyroid-fed animals 
of 13°11 mg. per 100 grm. body-weight, or nearly 76 per cent. 

In the first series of older animals the increase is 78 per cent.; in the 
second series of young animals the increase is 76 per cent. This increase in 
the weight of the suprarenals estimated per 100 grm. body-weight appears 
‘ to result from feeding the animal over a period of from 20 to 30 days 
with small doses of fresh ox thyroid. ‘The increased size in the thyroid- 
fed animal is at once apparent when the suprarenals are exposed. The 
increase appears to be partly of cortex and partly of medulla. 


Effect of Thyroid-Feeding on the Suprarenals 53 

According to Carlson, Rooks, and M‘Kie (5) the toxie effects of 
thyroid-feeding are partly ascribable to excess of protein, but only so 
when the dose is excessive. This is certainly not the case with amounts 
of from 0-2 to 0°5 grm. fresh thyroid daily. In the second series of rats 
an equivalent amount of raw ox flesh was given daily to the control 
animals (Table LIL). It does not appear to have caused any departure from 
the normal. 

TaBie II].—Seconp Series. Norma Rats or THE SAME AGE FED ON BREAD 

AND MILK +0°2 Grau. Fresa# Ox FLESH DAILY. 


6a |i i 8. 9. 10. 


| 
ae 3 | 4 | 5 | | 11. 12 
: = i a Le. (2c Bas, A= we sar 
& S& 3: |. a Bo, |e. tae | . se ete 
SiH | we \FE\SEIeE SE SEP| 4.8 S| eR) Ges | eas 
Meeeions jal MH b | shies | Sq | aoe | sae 
S Se jer | ss (e5| 85 se" | oe hF ) a7 | ssa! sks. 
Som |& |F |& les | 388 3 aus | sa 
a om a eh ee eee 
1 | M. | 23 days} 63 | 130 | +67 178 25 19°23 0034 0°026 07136 
a) Mm.) 94 , 73 | 147 | +74 186 24 16°32 0°039 0026 0°162 
espe. | 24s, 80 | 145 | +65, 184 24 16°55 0059 —(0':041 0246 
ieee. | 26° ,, 75 | 144 | +69 184 25 17°36 0057 0°039 0'228 
5 | M./ 26 ,, 74 | 148 | +74 187 25 16°89 | 0°052 0°035 0°208 
Averages 73 | 143 |+70 184 25 17°27 |0°048 | 0033 | 07196 


THE ADRENALIN CONTENT OF THE SUPRARENALS. 


~The amount of adrenalin in the suprarenals of the normal rat shows 
considerable variation. As a general rule it increases with the age and 
weight of the animal. The larger the suprarenals the more adrenalin do 
they contain, but exceptions are not infrequent. In the first series of 
normal rats (Table II.) the average adrenalin content is 0:065 mg., the 
lowest amount being 0:031 mg. in the second animal, and the highest 
0110 mg. in the last and largest rat. The amount of adrenalin per 
100 grm. body-weight is more constant, and averages 0°034 mg. for the 
thirteen rats. The percentage amount in the suprarenals averages 0224. 

In the second series of normal animals the average adrenalin content is 
0:048 mg., which expressed in mg. per 100 grm. body-weight is 0°033 mg., 
a figure which agrees very well with the average 0-034 mg. of the first 
series. The percentage in the suprarenals in this series is lower, and 
averages 07196. 

The administration of thyroid increases the adrenalin content. In the 
first series of rats (Table II.) the adrenalin content of the suprarenals 
averages 0°088 mg. as the result of feeding with thyroid. The lowest 
content of the series is 0057 mg. in the third animal, and the highest 
0-138 mg. in the thirteenth rat. The amount of adrenalin per 100 grm. 


54 Herring 


body-weight shows greater variation than in the normal, the lowest figure 
being 0-032 mg. in the eighth rat, and the highest 0-070 mg. in the seventh 
and tenth. The average is 0°052 mg., which, compared with the average 
normal amount of 0-034 mg., shows an increase of 0:018 mg. per 100 grm. 
body-weight, or nearly 53 per cent. 

In the second series of thyroid-fed animals (Table IV.) the amount of 
adrenalin is similarly increased. The average adrenalin content is 0-067 
mg. compared with 0:048 mg. in the normal. Per 100 grm. body-weight 
the adrenalin is 0:049 mg. as against 0°033 mg. in the normal. This is 
an increase in the adrenalin of the thyroid-fed rats of 0-016 mg., or nearly 
50 per cent. 


TABLE IV.-—Seconp SERIES. RATS OF THE SAME AGE FED ON BREAD 
AND MinK +0:2 Grou. FresH Ox THyRroIpD DAILY. 


Dale. 28.0 i] ody. | Bex) ER Maen 2, | ROS gel iglOs arate 12 
| | ares _— 
| | 2 eee ee eee 
| [ieee et arf eae feta) ea esr es RiSlae I IMS Tee cea wu |lize 3 A || one 
aes | SS ores SOC si Our seo seoe ee mecllenes ae =i) oD Ts 
| 8 l.—wGge |FS(O8 | Ba eo 2 co. Be. | 2) Sisco eee 
a | Si Sea [SmlF al China| FSe) Cee oe | a eee 
\2/7 | SB |Se(eS | 8s | Ss (Se8| Se me | 5k | see | see 
~ 7 | no a7 an a — ie = | oOrS — | Ea (SB) | = = =r 
 § aa CO lm |e 23s ee 33 |e 
1 | M.| 23 days| 60 | 103 |+43| 168 | 36 3495 0046 0-045 | 0-128 
Bi Me o4 | 67 | 131 eed ay7 | 32 24°42 | 0055) 0:042 | 0-172 
3| M./24 , | 86 | 158|+72|189| 60 38:00 | 0-088 0-056 0147 
4|M.|26 ,, | 76 | 141 |+65/183| 46 32°62 | 0-083 0:059 — 0°180 
5) M1} 26 ,, | 77 | 145 |468) 188 | 32.| 22:06 | 0-064 3-044 — 0200 
ie = eee Ae : Saale ae : 
Averages . . | 73 | 185 +62/181| 41 | 3038  0:067/ 0-049 0165 


In both series of thyroid-fed animals the percentage amount of 
adrenalin in the suprarenals is reduced, the figures being 0:191 against 
0224 normal in the first series, and 0°165 against 0°196 normal in the 
second series. The adrenalin increases with the increase in size of the 
suprarenals, but not to the same extent. It would seem, therefore, that 
there is a relatively greater hypertrophy of the non-adrenalin-forming 
tissue in the suprarenals. The animals examined received thyroid until 
the day before they were killed. If the increase in adrenalin is com- 
pensatory to the increased{intake of thyroid, as appears most probable, it 
is not unlikely that the suprarenals would show an even higher adrenalin 
content had the thyroid-feeding been discontinued for a longer interval 
before the animals were killed. One animal in each of the thyroid-fed 
groups died some time before examination. Both of these had the 
heaviest suprarenals recorded, viz. 60 mg. One of them, No. 3 in Table IV., 
was not examined until a considerable time had elapsed, and a certain 
amount of its adrenalin must have disappeared in the interval. From 


Effect of Thyroid-Feeding on the Suprarenals 55 


naked-eye examination the cortex of the suprarenals appears enlarged 
in the thyroid-fed rats, and it is possible that the cortex undergoes a 
relatively greater hypertrophy than the medulla. 

In a previous paper the author gave the average adrenalin content of 
the suprarenals per kg. body-weight as 0400 mg. in the rabbit and 
0:229 mg. in the cat. The adrenalin content of the rat more nearly 
approaches that of the rabbit than that of the cat, being 0-034 mg. per 
100 grm. body-weight. 


THE ADRENALIN CONTENT OF RETROPERITONEAL TISSUE. 

Attempts were made to estimate by Folin’s method the adrenalin in 
extracts of retroperitoneal tissue in the first six animals both of the control 
and thyroid-fed groups. The colour produced, however, was generally 
too weak for satisfactory measurement. Swale Vincent (17) was unable 
to find any chromaphil bodies in the rat. Fulk and Macleod (10) have 
recently affirmed the presence of retroperitoneal chromaphil tissue in the 
rat, and find that its extracts act similarly to weak solutions of adrenalin 
on intestinal muscle. The author, like Swale Vincent, has not been 
able to find any definite chromaphil bodies by histological methods. 
Extracts of the retroperitoneal tissue, when carefully prepared so as to 
precipitate the proteins, give a variable response to Folin’s reagent. 
Sometimes the colour produced is very slight, at other times quite measure- 
able. A slight reaction to Folin’s reagent does not necessarily indicate the 
presence of adrenalin. The stronger reaction, when present, is probably 
due to the inclusion in the retroperitoneal tissue of accessory suprarenals. 
They are not uncommon in the rat, and are often very minute. It is often 
impossible to distinguish them by anything but microscopical observation. 

The accessory suprarenals appear to be larger, and their chromaphil 
cells more abundant in rats which have been thyroid-fed. In several of 
the animals recorded in Table II. the amount of adrenalin in the accessory 
suprarenals must have been quite appreciable. This was especially the 
case in rat No. 13, in which the suprarenals contained 0:138 mg. of 
adrenalin. Subsequent investigation of the retroperitoneal tissue of this 
animal after fixation in bichromate of potash solution showed the existence 
of three small accessory suprarenals each containing chromaphil cells. 
The total adrenalin content of this animal would have been larger if these 
bodies had been included. 

While no definite figures can be given, the author believes that there 
is evidence to show that where accessory suprarenals are present they 
participate with the suprarenals in the enlargement and in the increase 
of adrenalin content resulting from thyroid-feeding. 


MORTALITY UNDER SMALL Doses OF THYROID. 
Reference has been made to the frequency of sudden death occurring 
in rats while under the influence of small doses of thyroid. This has 


56 Herring 


happened for the most part to animals which were putting on weight 
rapidly, and were in apparently good condition. In all cases of the kind 
examined the suprarenals are hypertrophied. The most striking feature, 
however, is the increased size of the heart. In No. 3 rat (Table IV.) the 
weight of the heart is nearly treble that of the normal animal of the same 
body-weight, and the age of the animal is less than three months. Ex- 
amination of the heart was unfortunately overlooked in the earlier experi- 
ments recorded, but in the later ones the heart is found to be considerably 
hypertrophied in the thyroid-fed animals. The increase in weight and in 
the muscularity of the left ventricle are most striking features. Further 
observations are being made on this condition. 


SUMMARY OF CONCLUSIONS. 


The administration of small quantities, 0-2 to 0°5 grm., of fresh ox 
thyroid daily to white rats increases the size and weight of the suprarenals 
both in young and adult animals. Reckoned in mg. per 100 grm. body- 
weight, the extent of the increase in the eighteen male animals examined 
averages a figure which is equivalent to arise in weight of from 76 to 78 
per cent. above the normal. This is the result of feeding the animals with 
the above-mentioned doses of thyroid for a period of from three to four 
weeks. 

Both cortex and medulla participate in the enlargement, but the hyper- 
trophy of the cortex is somewhat greater than that of the medulla. 

There is reason to believe that where accessory suprarenals are present 
they too undergo enlargement. 

The adrenalin content of the suprarenals of the white rat increases 
as a general rule with increase in weight of the animal. The amount 
of adrenalin normally present is about 0°034 mg. per 100 grm. body- 
weight. 

Feeding with thyroid increases the adrenalin content of the suprarenals. 
The average amount in the eighteen animals examined averages from 
0049 to 0°052 mg., which is equivalent to a rise in weight of about 50 
per cent. 

Owing to the relatively greater increase in weight of the suprarenals, 
resulting from thyroid-feeding, the percentage amount of adrenalin in them 
is decreased, though only to a small extent. . 

It is probable that the adrenalin in the accessory suprarenals, when 
these are present, is also increased. 

Rats fed with small doses of thyroid not infrequently die suddenly 
when apparently thriving. In the animals examined which have thus 
died there is great increase in the weight of the suprarenals and in the 
adrenalin content. The heart is greatly hypertrophied in addition. There 
is evidence that thyroid-feeding results in rapid cardiac hypertrophy in 
white rats. The condition is being further investigated. 


Effect of Thyroid-Feeding on the Suprarenals 57 


The author has to thank Mr Niven of Strathkinness for his kindness 
in ensuring a regular supply of fresh ox thyroids. 


The expenses of the research have been met by a grant from the 
Carnegie Fund. 


REFERENCES. 
(1) Asner and Frack, Zeitschr. f. Biol., 1910, lv. 83. 
(2) Brokine and TrenpELENBURG, Deutsch. Arch. f. klin. Med., 1911, ciii, 168. 
(3) Cannon and pE ta Paz, Amer. Journ. Physiol., 1911, xxviii. 64. 
(4) Cannon and Hoskixs, Amer. Journ. Physiol., 1911, xxix. 274. 
(5) Cartson, Rooks, and M‘Kre, Amer. Journ, Physiol., 1912, xxx. 129. 
(6) Donatpson, The Rat, Philadelphia, 1915. 
(7) Etutorr, Journ. Physiol., xliv. 374. 
(8) Fotry, Journ. Biol. Chem., 1912-13, xiii. 477. 
(9) Fragnxker, Arch. f. exper. Path., 1908-9, Ix. 395. 
(10) Furk and Macteop, Amer. Journ. Physiol., 1916, xl. 21. 
(11) Herrine, Quart. Journ. Exper. Physiol., 1916, ix. 391. 
(12) Hewrrt, Quart Journ. Exper. Physiol., 1914, viii. 297. 
(13) Hoskins, Journ. Amer. Med. Assoc., 1910, lv. 1724. 
(14) Krause, quoted from Schafer (16). 
(15) Orr and Scort, Journ. Pharm. and Exper. Therap., 1911-12, iii. 625. 
(16) ScuArer, The Endocrine Organs, London, 1916. 
(17) Swate Vincent, Internal secretion and the ductless glands, London, 1912. 


CARBOHYDRATE METABOLISM IN RELATION TO THE THY- 
ROID GLAND. II.: THE EFFECT OF THYROID-FEEDING 
ON THE GASEOUS METABOLISM. By W. Cramer and R. 
M‘Catu.! (From the Imperial Cancer Research Fund, London, and 
from the Physiology Department, Edinburgh University.) (With 
eleven figures. ) 


(Received for publication 2nd September 1916.) 


INTRODUCTION. 


THE following investigation was carried out in continuation of previous 
studies on the influence of the thyroid secretion on carbohydrate meta- 
bolism. Cramer and Krause (1) had found that feeding with thyroid 
gland produced after two or three days a complete disappearance of 
glycogen from the liver, even when the animals (rats and cats) were kept 
on a diet rich in carbohydrates (e.g. bread and milk). It was further 
noted that under these conditions no sugar appeared in the urine, whereas 
glycosuria would be expected on the basis of the current conception of 
carbohydrate metabolism. 

There are three possibilities which might account for this apparently 
paradoxical condition of carbohydrate metabolism. Either the suspension 
of the glycogenie function of the liver is compensated for by an increased 
deposit of glycogen in the muscle—and a slight increase so far as absolute 
amounts are concerned would be sufficient to account for the amount of 
carbohydrate normally deposited in the liver. Or, secondly, one might 
postulate for the thyroid hormone a direct stimulating effect on the oxida- 
tion of carbohydrates, so that the disappearance of glycogen from the liver 
would have to be interpreted as an effect secondary to the increased oxida- 
tion of carbohydrates. Or, thirdly, carbohydrates might be transformed 
into and deposited as fat. This last possibility can be dismissed at once, 
since the rapid disappearance of fat under the influence of thyroid feeding 
is a well-established fact: The first possibility that the muscles act 
vicariously for the liver as glycogen depots was tested by R. A. Krause (2) 
by determining the glycogen content of the muscles of thyroid-fed 
animals. Since in such animals the glycogen percentage of the muscles 
is not increased, this possibility can be excluded. The second and last 
possibility, which depends on the assumption that the thyroid secretion, 
acting as a true hormone in Schiafer’s definition of the term (3), directly 


1 Carnegie Research Scholar, 1914-1915. 


60 Cramer and M‘Call 


stimulates the oxidation of carbohydrates, has already been considered by 
Cramer and Krause and was excluded by them on the ground that 
glycogen is absent from the liver even immediately after a meal rich in 
carbohydrates, whereas one would expect to find only a more rapid dis- 
appearance of glycogen from the liver of thyroid-fed animals and a 
replenishing after a meal if the effect on the liver glycogen were merely a 
secondary one. The conclusion that this possibility must be excluded is 
confirmed by other facts. If the thyroid hormone produced a direct 
stimulating effect on the oxidation of carbohydrates, one would expect that 
hyperfunction of the thyroid gland, whether produced experimentally by 
thyroid-feeding or studied clinically in Graves’ disease, should produce a 
rise in the limit of assimilation for sugar. It is well known that in Graves’ 
disease this is not the case; on the contrary, there is frequently a tendency 
to glycosuria in that condition. The effects of thyroid-feeding which have 
been studied in dogs show a slight but distinct lowering of the limit of 
assimilation for glucose. Conversely, one would expect that thyroid 
insufficiency, whether studied clinically in myxcedema or _ produced 
experimentally by extirpation of the gland, should lead to an impaired 
oxidation of carbohydrates and therefore a lowering of the limit of 
assimilation for glucose, if not actually a diabetes mellitus. Here again it 
is well known in the ease of myxcedema that this is not the case, but that 
there is a tendency in the reverse direction. The condition of the carbo- 
hydrate metabolism in thyroidectomised animals, of which little is known, 
has been studied by us in rats and may be dealt with in a subsequent 
paper. It is sufficient to state here that our observations on the respiratory 
quotient in thyroidectomised animals give no evidence of an impaired 
power of oxidation of carbohydrates in thyroidectomised animals. This 
is in agreement with the fact that thyroidectomy has never been found to 
produce a diabetes mellitus, and that clinically diabetes has sometimes been 
found to develop in persons suffering from Graves’ disease, but not in 
myxcedematous patients (4). 

Moreover, if the thyroid secretion increased or facilitated the oxidation 
of carbohydrates the thyroid hormone should represent the long-sought-for 
remedy against diabetes mellitus. The literature contains no information 
on this point. Our own experience is limited to one case of diabetes 
mellitus, in which the condition was greatly aggravated by the administra- 
tion of thyroid gland. 

The condition of the carbohydrate metabolism in a state of experimental 
hyperthyroidism as induced by thyroid-feeding is therefore one which 
cannot be accounted for by the orthodox conception of carbohydrate meta- 
bolism. We must therefore, in looking for an explanation, dispense with 
the guidance offered by that theory and try to approach the problem 
without the bias of a theory. 

Since in a state of experimental hyperthyroidism carbohydrates are 
neither deposited as such nor excreted as such, nor transformed into fats, 


Carbohydrate Metabolism in relation to the Thyroid Gland 61 


we must arrive per exclusionem at the conclusion that they are oxidised. 
Or we must, assume the existence of a hitherto unrecognised depot of 
carbohydrates functioning in a manner entirely ditferent from the recog- 
nised depdts in the liver and the muscles. The following observations 
were made in order to see whether this conclusion of an increased 
oxidation of carbohydrates as the result of thyroid-feeding can be 
demonstrated experimentally. 

It must be clearly realised that such an increased oxidation of carbo- 
hydrates could not be due, for reasons which have been given above, to a 
stimulating effect of the thyroid hormone on the oxidative processes in 
the cells, but would have to be looked upon as the result of the inhibition 
of the glycogenic function of the liver. This, it is true, runs counter to 
all accepted conceptions of carbohydrate metabolism ; it is, in fact, the very 
reverse of what one would expect to find. The problem is therefore of 
interest not only in relation to the action of the thyroid secretion, but 
raises the wider question of the correctness of our current conceptions of 
carbohydrate metabolism. 


EXPERIMENTAL PART. 


An increased oxidation of carbohydrates uncomplicated by any other 
changes in metabolism is not known to give rise to any very obvious 
manifestations. It is therefore not so easily recognised or demonstrated : 
not so easily, for instance, as the reverse condition of a diminished 
oxidation of carbohydrates. Under the simplest conditions one might 
expect to find that the respiratory quotient should maintain itself after a 
meal containing carbohydrates for a longer time at the initial high level 
which indicates an oxidation of carbohydrates and which is due mainly to 
increased excretion of CO,. This was the plan which was followed in 
this investigation. It may be pointed out at once, however, that that 
expectation can be fulfilled only if, as stated above, the increased oxidation 
of carbohydrates is uncomplicated by any other changes in metabolism. 

The observations were made on rats. These animals, being omnivorous, 
are especially suitable for such observations. The estimation of the 
gaseous metabolism was made by the method of Haldane and Pembrey, 
which is well adapted to observations on small animals. Some modifica- 
tions were found to be an improvement. Instead of using pumice-stone 
saturated with concentrated sulphuric acid for the absorption of water, the 
current of air was passed through concentrated sulphuric acid in a squat 
wash bottle constructed on the principle of Folin’s apparatus for the 
absorption of ammonia. The carbonic acid was absorbed by a tube 
containing first a pad of glass wool soaked in strong caustic soda and then 
soda lime. The precautions recommended by Haldane and Pembrey 
(weighing against dummy tubes, controls, etc.) were observed. Since with 
the short hourly periods of observation adopted in these experiments the 
excretion of CO, and H,O amounted only to a few decigrammes, it was 


62 Cramer and M‘Call 


essential to have a balance which would indicate weights of about half a 
kilo (the weights of the animal chamber, absorption tubes, which were all 
made of thin glass) accurately and rapidly to a milligramme. Such a 
balance was obtained from Collot (Paris). It is capable of weighing a 
kilo accurately to fractions of a milligramme, and by means of a damper 
arrangement the weighing of the animal chamber can be carried out in 
about two minutes. The observations on the respiratory exchange were 
made in hourly periods from the third to the eighth hour after a meal. 

In a series of preliminary observations the conditions were established 
under which it is possible to obtain, in the case of normal animals, constant 
results which can serve as a standard. The animals, which were of 
approximately the same age and weighed from 125 to 160 grammes, were 
kept in the specially constructed metabolism cages devised by Schafer (5), 
in which the amount of food eaten by the animals can be accurately 
determined. The animals were fed at regular hours with weighed 
quantities of bread and milk (equal parts). In the evening the food was 
placed in the cages at 6 p.m. and removed at 8 p.m., when the amount of 
food consumed was noted. Although in the evening the animals were 
allowed to eat ad libitum, the amount of food consumed varied only 
within narrow limits from day to day. The animals were fed again in the 
morning at 8.45 a.m., when 10 grammes of the bread-and-milk mixture 
were offered to them, and the empty beakers were removed after an hour. 
Observations on the gaseous metabolism were made only if the animals 
had been kept for at least ten days under these conditions. The animals 
rapidly became accustomed to the routine, and ate the food with avidity 
when it was placed in the cages. The observations on the gaseous 
metabolism were begun as a rule at 10.45 a.m. and lasted till 4.45 p.m. 
Observations were made only if the animals had completely eaten the 
10 grammes of bread and milk within one hour, which they did almost 
invariably. 

Under these conditions the period of observation begins with a 
respiratory quotient of about 1 in the third hour after feeding (ie. from 
10.45 a.m. to 11.45 a.m.) and falls gradually till in the eighth hour after 
feeding it reaches a quotient of about 0°75. The observations were made 
in a warm room, which could be kept at an even temperature not only 
through the day but also throughout the whole year. The animals were 
kept in an adjoining room at about the same temperature. 

The animals when in the respiration chamber remained quiet, and were 
asleep during almost the entire period of observation, except during the 
weighing at the end of each hourly period and during the last (eighth) 
hour when, being hungry, they became slightly restless and began to move 
about. The general condition of activity or inactivity of the animals 
during the periods of observation was noted. The significance of this 
point will be discussed below. 

When it was desired to induce experimental hyperthyroidism, dried 


Carbohydrate Metabolism in relation to the Thyroid Gland 63 


thyroid gland in the form of a powder was mixed with the food in the 
morning and evening. Half a gramme of dried thyroid was given each 
time. In some instances half a lobe of a fresh sheep's gland was given, 
instead of half a gramme of the dried thyroid, during part of the 
experimental period. The dried thyroid powder had been prepared 
recently in the laboratory from fresh glands obtained from the slaughter- 
house. It is in our experience essential for experimental purposes to use 
a thyroid preparation made recently from fresh thyroids, and under 
personal supervision, if one wishes to obtain reliable and constant results. 
It was noted as an important point that the thyroid-fed animals were not 
more active during the periods of observation than the normal animal. 
They also were asleep during almost the entire period of observation, 
except the last hour. 

The results are represented graphically in figs. 1-ll. Figs. 1-6 
represent the curves of the respiratory quotients from the third to the 
eighth hour after a meal of 10 grammes of bread and milk. In these 
figures the interrupted lines labelled “N” represent the results obtained 
with the normal animal before the administration of thyroid gland. The 
curves obtained on different days are labelled N,, N,, and N, respectively. 
The solid black lines labelled “Th.” represent the results obtained in 
animals fed with thyroid gland. ‘The uumbers attached to the letters 
~ Th.” indicate the number of days during which thyroid-feeding has been 
maintained. Thus “Th.5” means that the animal has been fed with thyroid 
gland for the five days preceding the observations, the last dose being 
given with the meal immediately preceding the observations. The dotted 
lines labelled “R” represent results obtained in animals recovering from 
the effect of thyroid-feeding. Here again the number attached to the 
letter “ R” indicates the number of days after the last dose of thyroid has 
been given. 

Figs. 7-11 give graphically the amounts of CO, excreted and O, 
absorbed in milligrammes in hourly periods. The upper solid lines re- 
present CO, excreted in milligrammes, the lower dotted lines represent the 
O, absorbed in milligrammes. The total amount of CO, excreted during the 
whole six-hourly period is given in milligrammes underneath each curve. 

Further experimental details are given in the protocols appended to 
this paper. 

The results obtained with the four rats in the normal state (before the 
administration of thyroid gland) may be considered together. They 
agree in every respect with the observations of Pembrey and Spriggs (6) 
on the gaseous metabolism of normal rats. After a meal rich in carbo- 
hydrates the CO, excretion shows a marked rise and then gradually falls. 
At the same time the oxygen excretion remains fairly constant. The 
changes in the respiratory quotient are therefore mainly dependent upon 
the changes in the CO, excretion: the quotient reaches its maximum in 
the third hour after a meal, rising to about unity, when carbohydrates are 


64 Cramer and M‘Call 


being oxidised almost exclusively, and then gradually falls, until it reaches 
in the eighth hour the level of the fasting animal with a quotient .of 
about 0°75, indicating the almost exclusive combustion of fat. Move- 
ments of the animal increase both the CO, excretion and the O, absorption, 
so that any restlessness of the animal during the period of observation 
will produce slight irregularities in the curves of the CO, excretion and 
O, absorption, whereas the respiratory quotient remains unaffected. 
These irregularities, resulting in a slight increase in both CO, excretion 
and O, absorption, can be noted in several experiments, especially during 
the last hour of the period of observation, when, as stated above, the 
animals were slightly restless from hunger. 

The effect produced by thyroid-feeding shows two distinct phases, 
which may be called the “early” and the “later” stages respectively of 
experimental hyperthyroidism. In our experiments the early stages were 
observed during the first two or three days of thyroid-feeding, the later 
stages, from the third to the sixth day. It must be understood that this 
“later” stage, which will be discussed here, does not refer to the extreme 
effect which can be produced by prolonged feeding for several weeks with 
large doses of thyroid gland, and which leads to enteritis, great emaciation, 
listlessness, and eventually death. In our experiments thyroid-feeding 
was never carried to this extreme stage, and during the entire period of 
observation the animals remained in good condition: they were lively, ate 
their food with avidity, and the feces were well formed. With Rat 1 
observations were made in the “later” stage, with Rat 2 in the “early” 
stage of thyroid-feeding, while in the case of Rat 4, and especially of Rat 3, 
the observations extend over both stages. The observations on Rat 3 may 
therefore be considered first and in some detail. 

During the “early ” stage, which in the case of Rat 3 extends over the 
first two days of thyroid-feeding, the hourly curve of the respiratory 
quotient resembles that of the normal animal. There is a rapid rise 
immediately after a meal and a slow fall. There are, however, certain 
significant differences. The post-prandial rise is maintained for a longer 
period; indeed, on the first day of thyroid-feeding the respiratory quotient 
is higher in the fourth hour than it is in the third, whereas in the 
normal animal the quotient obtaining in the third hour is never exceeded 
at any subsequent period. There is a similar irregularity during the 
eighth hour, the last respiratory quotient being higher than that of the 
preceding (seventh) hour. In the case of Rat 3 the curve for the thyroid- 
fed animal lies at a higher level than that for the normal, but that, as 
will be seen, is not invariably the case. These changes in the respiratory 
quotient curve are due entirely to corresponding changes in the CO, 
excretion. The post-prandial rise in the CO, excretion is more prolonged 
in the thyroid-fed animal. The O, absorption remains fairly constant, and 
is not markedly atfected. The total CO, excretion during the entire 
period of observation is slightly increased, but it is not so much the in- 


Carbohydrate Metabolism in relation to the Thyroid Gland 65 


crease in the total CO, excretion as the distribution over the various 
periods that is significant. All these observations point clearly to the 
conclusion that during the “early” stage of experimental hyperthyroidism 
more of the ingested carbohydrates are oxidised than in the normal 
animal. 

The same condition of the gaseous metabolism during the early stage 
of experimental hyperthyroidism is seen in the experiments with Rat 2: 
the prolonged post-prandial rise of the CO, excretion is especially notice- 
able on the third day of thyroid feeding. In the case of Rat 4 the observa- 
tions on the normal animal gave the least uniform results, and the first two 
days of thyroid feeding show no obvious change from the normal, either 
as regards the curve of the respiratory quotient or that of the CO, excre- 
tion. The latter is slightly increased, but the quotient curve is actually a 
little below the normal. There is only the characteristic tilt-up of the curve 
in the eighth hour after a meal. which is noticeable in all the three cases of 
“early ” stages of experimental hyperthyroidism which we have investigated. 

In the “later” stage of experimental hyperthyroidism the gaseous 
metabolism undergoes a complete change, and the interpretation of the 
results becomes more difficult. The most obvious change is a general 
marked increase in both the CO, excretion and the O, absorption, and 
this is the effect which has so frequently been described as the result of 
thyroid administration. The explanation usually given—if it is an ex- 
planation—is that the thyroid hormone increases the processes of oxida- 
tion. The further analysis of this increased gaseous exchange by hourly 
observations shows that there is not only a quantitative but also a quali- 
tative change. The curve of the CO, excretion retains its characteristic 
features, in fact the post-prandial rise is even more marked. But the O, 
absorption, instead of remaining more or less constant, now also shows a 
pronounced post-prandial rise and subsequent fall. ‘The two curves which 
in the normal animal are convergent! run parallel. The respiratory 
quotient curve thus becomes flattened out, although it still shows a 
maximum. But this maximum does not now occur in the third hour after 
feeding at the beginning of the experiment as in the normal animal, but 
is delayed. In Rat 4 with three days’ feeding it is found in the fourth 
hour, in Rat 1 with five days’ feeding in the sixth hour, and in Rat 3 with 
five days’ feeding in the seventh hour, so that in this last case the curve 
for the respiratory quotient becomes inverted. A notable point is that the 
respiratory quotient curve may be at a high level as in Rat 1 (first series) 
and Rat 4, or at a low level as in Rat 1 (second series) and Rat 3. 

The high-level curves, where the respiratory quotient remains above 
0-9 from the third to the seventh hour, clearly indicate an increased oxida- 
tion of carbohydrates. But that explanation does not account in itself for 
the great increase in the total gaseous exchange. And at first sight it does 
not seem possible to reconcile this explanation with the low-level curves, 

1 This applies to the curves of the weights of CO, and O,. 


VOL. X1., NO. 1.—1917. = 


66 Cramer and M‘Call 


where, as in the case of Rat 3, the quotient never rises even as high 
as 0°85. 

Such abnormally low quotients, obtained after a meal rich in carbo- 
hydrates, cannot be explained in this case as being due to an impaired 
oxidation of carbohydrates. We know that thyroid feeding prevents the 
storage of carbohydrates in the liver. If in addition to the inability to 
store carbohydrates there were also an impaired oxidation of carbo- 
hydrates, experimental hyperthyroidism ought to produce a_ severe 
diabetes mellitus. But, as has been mentioned in the Introduction, and 
as we have repeatedly tested in the course of these experiments, it does 
not even produce a glycosuria. Another reason for excluding an impaired 
oxidation of carbohydrates as a possible explanation is that it would be 
almost impossible to understand how thyroid feeding could, under 
apparently identical experimental conditions, produce two diametrically 
opposed conditions, even in the same animal (Rat 1 at different times). 

One must therefore look for a different explanation of the lowering 
of the quotient when it occurs. The only other known cause which 
could account for it under the conditions of our experiments is the 
transformation of protein and possibly fat into carbohydrate. These 
processes lower the quotient by increasing the absorption of O,. The 
carbohydrate thus formed from protein and fat is not deposited, but at 
once oxidised. 

The increased breakdown of protein and disappearance of fat which 
this explanation presupposes is a well-known and long-established effect 
of thyroid-feeding. It appears on our interpretation as a secondary 
result of the action of the thyroid secretion on the glycogenic function 
of the liver. The more protein or fat is transformed into carbohydrate 
and then oxidised, the more the curve of the respiratory quotient will 
be depressed. A low-level curve thus indicates that a relatively large 
amount of protein or fat is being transformed into carbohydrate and 
oxidised as such, while a high-level curve signifies that the carbohydrate 
which is being oxidised is mainly that pre-existing in the food or still 
present in the organism. 

The correctness of the explanation which has been given can be tested 
by the observations recorded in this paper. The greater the amount of 
protein or fat transformed into carbohydrate, the greater will be the amount 
of carbohydrate undergoing oxidation. The measure of the first-mentioned 
process is the level of the respiratory quotient curve: the lower the level 
the greater the amount of protein or fat undergoing transformation. The 
measure of the second process is the increase in the CO, excretion above 
that of the normal animal. One should therefore expect to find that the 
lower the level of the respiratory quotient curve the higher the level of 
the CO, excretion. <A glance at the figures shows that this is the case. 
In Rat 1, second series, and Rat 3, where the respiratory quotient curve 
is very low, the increase in the CO, excretion is much greater than in 


Carbohydrate Metabolism in relation to the Thyroid Gland 67 


Rat 1, first series, or Rat 4, where the level of the respiratory quotient 
curve is high. Another suggestive fact is that under the conditions of 
these experiments the later stages of experimental hyperthyroidism appeared 
on the third day, with the exception of Rat 2; and under the same con- 
ditions it is on the second or third day that the glycogen has completely 
disappeared from the liver. 

When thyroid feeding is discontinued the gaseous metabolism rapidly 
returns to the normal. 

In the interpretation of the low quotients given above it has not been 
overlooked that a process involving the transformation of, say, protein 
into carbohydrates and the subsequent oxidation of the carbohydrate 
thus formed would, if considered in toto, give the same respiratory 
quotient as the direct oxidation of the same amount of protein. One 
might therefore be inclined to ask whether the interpretation which has 
‘been given is not merely a complicated way of expressing the fact that 
thyroid feeding produces an increased oxidation of carbohydrates, protein 
and fat, and whether it would not be a simpler and equally adequate 
explanation of the phenomenon to assume that the thyroid hormone 
increases all the oxidative processes of the cell / 

It is important to realise that such an explanation differs toto coelo 
from the one given in this paper, in which the various changes in meta- 
bolism are looked upon as secondary effects which can be traced back to 
the primary fact of the inhibition of thefglycogenic function of the liver. 
The alternative explanation of an increased oxidative power of the cell 
due to the thyroid hormone—a statement which in reality is not so much 
an explanation as a paraphrase—interprets these changes as direct primary 
effects. If an increased power of oxidation on the part of the cell were 
the cause, the curve of the respiratory quotient of a thyroid-fed animal 
should be essentially similar to that of a normal animal, while the curves 
for the CO, excretion and O, absorption should run parallel to those of 
the normal animal, but on a higher level. Moreover, it has already been 
shown in this paper that the increased oxidation of carbohydrates cannot 
be ascribed to a primary effect. We would therefore have to look upon 
the increased oxidation of fat and protein as a separate and entirely 
unrelated effect, which is superimposed upon the effect of the autacoid 
of the thyroid gland on the liver glycogen and the resultant increased 
oxidation of carbohydrates. Even then we should be unable to account for 
the changes which the curves of the respiratory quotient and of the Oz 
absorption undergo in the later stages of experimental hyperthyroidism. 

There are, moreover, several facts which point to the breakdown of 
protein being secondary to the effect on the carbohydrate metabolism in 
experimental hyperthyroidism. ‘The first is that the transformation of 
protein into carbohydrate occurs also in another condition associated with 
the disappearance of glycogen from the liver, namely, in severe diabetes 
mellitus. The difference is that in the latter condition the carbohydrate 


68 Cramer and M‘Call 


formed from protein is excreted in the urine as sugar, while in experi- 
mental hyperthyroidism it is oxidised. Another point in favour of the 
interrelationship is the condition of the nitrogen distribution in the urine 
after thyroid feeding. If the thyroid hormone affected the protein meta- 
bolism directly, one would expect, as Cramer and Krause (7) pointed out, 
an increased excretion, especially of the end-products of endogenous protein 
metabolism. But this expectation is not fulfilled: the excretion of these 
substances remains unchanged; instead, the picture of the urinary nitrogen 
distribution is very similar to that obtained when carbohydrates are removed 


from the diet. 
CONCLUSIONS. 

Observations on the gaseous metabolism in experimental hyper- 
thyroidism confirm the conclusion arrived at on other grounds, that the 
thyroid secretion produces an increased oxidation of carbohydrates. This 
effect is not due to a direct stimulating effect of the thyroid secretion on 
the oxidative processes in the cell, but follows upon its action in discharg- 
ing glycogen from the liver. 

Two stages of experimental hyperthyroidism can be distinguished : 
an “early” stage in which the pre-existing carbohydrate of the food 
and of the deposits in the organism is oxidised. When all the glycogen 
has been removed from the liver a “later” stage sets in, in which there 
is in addition a formation of carbohydrate from protein and possibly also 
from fat, and a subsequent oxidation of the carbohydrate thus formed. 
The “later” stage is characterised by a marked rise in the CO, excretion 
and the O, absorption. 

The action of the thyroid secretion on the glycogenic function of the 
liver lies thus at the root of the increased oxidation of proteins, carbo- 
hydrates, and probably also of fats produced by thyroid feeding. 


APPENDIX. 


Protocols. 


In every case the animals were fed with 10 grm. of bread and milk 
from 8.45 a.m. to 9.45. Food removed at 9.45 a.m. Fed again from 6 p.m 
to 8 p.m. with bread and milk ad libitum. Food removed at 8 p.m. Esti- 
mations of the gaseous metabolism, when made, were begun at 10.45 a.m. 
and continued for the following six hours in hourly periods. 

As the main results obtained are represented graphically in figs. 1-11, 
only the following data concerning the experimental conditions under 
which the observations were carried out are abstracted from the protocols 
and given here to supplement the records given in the figures. 

Rat 1. First Series.—From 5/10/14 to 15/10/14 fed as described 
above. No observations on gaseous metabolism made. Same method of 
feeding continued, and observations made on— 


Carbohydrate Metabolism in relation to the Thyroid Gland O9 


16/10. Referred to in figs. as Exp. N1, wt. of rat 135 grm., temp. of room 16°. 

19/10. ‘3 : N2, a 135 grin., : 16°5 

20/10. zs = N3, . 135 grm., a 165°. 
Fed with half lobe of fresh sheep's thyroid from 21/10 to 25/10, morning 

and evening, in addition to bread and milk, and on morning of 26/10 at 

8.45 a.m. Observations made on— 

26/10. Referred to in figs. as Exp. Th. 5, wt. of rat 120 grm., temp. of room 16°. 


Thyroid-feeding discontinued after 26/10, 8.40 a.m. Fed with bread 
and milk as usual. Observation made on— 


27/10. Referred to in figs. as Exp. R1, wt. of rat 120 grin., temp. of room 16°. 

Rat 1. Second Series.—From 28/10 to 24/11 fed as described above. 
No observations on metabolism. Same method of feeding continued, and 
observations made on— 


25/11. Referred to in figs. as Exp. N1, wt. of rat 160 grm., temp. of room 18°. 
26/11. . :: N2, iM 160 grm., s 18°. 


Fed with 4 grm. dried sheep’s thyroid, in addition to bread and milk, 
on 26/11, at 6 p.m., and twice daily from 27/11 to 1/12 and on 2/12 at 
8.45 a.m. Observations made on— 

1/12. Referred to in figs. as‘Exp. Th.5, wt. of rat 150 grm., temp. of room 16°. 
2/12. 7 = 'PREAG, ie 150 grm., * lv". 

Rat 2.—From 7/10 till 20/10 fed as described above in detail. No 
observations made. Same method of feeding continued, and observations 
made on— 

21/10. Referred to in figs.as Exp. 
23/10. - f: : 

Fed with half lobe of fresh sheep’s thyroid on 27/10 at 5 p.m., 28/10 at 
8.45 a.m. and 6 p.m., with } grm. of dried thyroid gland on 29/10, 30/10, 
and 31/10, at 8.45 a.m. and 6 p.m., in addition to need and milk. Obser- 
vations were made on— 


29/10. Referred to in figs. as Exp. Th.2, wt. of rat 120 grm., temp. of room 16°. 


wt. of rat 140 grm., temp. of room 17°. 
135 grm., F 16°. 


42 


AE 
v2, 


30/10. Js i Th.3, “ 125 grm., : 14°. 
31/10. re Th.4, i. 115 grm., ) 14°5°. 


Thyroid-feeding discontinued from morning of 1/11. Fed as usual with 
bread and milk. Observations made on— 
2/11.! Referred to in figs. as Exp. R2, wt. of rat 115 grm., temp. of room 17”, 

Rat 3.—From 5/11 to 15/11 fed as described above. No observations 
made. Same method of feeding continued, and observations made on— 
16/11. Referred to in figs. as Exp. N1, wt. of rat 155 grm., temp. of room 15:5". 
17/11. F ¥ N2, a 155 grm., s Bis. 
18/11. y _ N3, i 155 grm., ; 16°: 


1 For three hours only, from 10.40 a.m. to 1.40 p.m. 


70 Cramer and M‘Call 


Fed with § erm. dried thyroid in addition to bread and milk on 18/11 at 
6 p.m. and twice daily thereafter from 19/11 to 22/11, and on 23/11 at 
8.45 a.m. Observations made on— 
19/11. Referred to in figs. as Exp. Th.1 wt. of rat 158 grm., temp. of room 17”. 


20/11. FF rs he? Ee 153 grm., ei ae 
21/11. " ” thes i 153 grm.., Ed Lite 
23/11. As 3 (TheES - 150 grm., .- 182. 


Thyroid feeding discontinued after 23/11, 8.45 a.m. Usual method of 
feeding continued, and observations made on— 
eee Referred to in figs. as Exp. R1, wt. of rat 150 grm., temp. of room Les 
PAG ALE < ss R4, x 150 orm., 17 
30/11. £ Es Rs, es 160 grm., e 18°. , 
Rat 4.—From 4/10 to 5/11 fed as described above with bread and 
milk. No observations made. Same method of feeding continued, and 
observations made on— 
6/11. Referred to in figs. as Exp. N1, wt. of rat 120 grm., temp. of room 20”. 


fled ss ; N2, 59 120 grm., ‘ 18s. 
9/11. 4 5 N3, % 125 grm.., M Les 
Fed with 4 grm. of dried sheep’s thyroid on 9/11 at 5 p.m., and twice 


daily thereafter from 10/11 to 12/11, in addition to bread and milk. 
Observations made on— 

10/11. Referred to in figs. as Exp. Th.1, wt. of rat 125 grm., temp. of room 18°. 
Uae oe ms The: et 120 grm., 2 Leis 
12/11. i e iPheS. 118 grm., . lites 


1 On 23/11 observations were begun an hour later than usual, and include therefore 
only the fourth till eighth hour after the meal. This is indicated in the figures. 


REFERENCES. 


(1) Cramer and Krause, Proc. Roy. Soc., B, 1913, Ixxxvi. 550. 

(2) Unpublished observations. 

(3) ScuArer, Endocrine Organs, Longmans, Green & Co., London, 1916, 5. 

(4) Detailed references to the literature on these points are given in the paper 
by Cramer and Krause quoted above. 

(5) E. A. Scorer, Croonian Lecture, Roy. Soc. Proc., B, 1909, lxxxi. 442. 

(6) Pemprey and Spriaes, Journ. Physiol., 1904, xxxi. 320. 

(7) Cramer and Krauss, op. cit. 


71 


~ 


Metabolism in relation to the Thyroid Gland 


Carbohydrate 


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Carbohydrate Metabolism in 1 


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MG.COz2 NI N.3 TH.S Ri 


200'— 
3142 MG.COz 3025 MG.CO, 3375 MG.COz 2959 MG.CO, 


Fic. 7.—Rat1. First Series. See also fig. 1. 
Curves of CO, excretion (solid black line) and 0, absorption (dotted line) in milligrammes. 


MG.CO, N.1 N.2 TH.5 TH.G 


800 


200 
3B2IIMG.COz 3375MGCO, 4213MG.CO, 4366MG.CO, 


Fic. 8.—Rat 1. Second Series. See also fig. 2. 
Curves of CO, excretion (solid black line) and O, absorption (dotted line) in milligrammes. 


NG. COz N.I N.2 TH.2 TH. TH. 4 2 


200 
3123 MG.COz 2858 MG.CO, 2940 MG.CO, 3346MG.CO, 3019 MG.CO, 


Fic. 9.—Rat 2. See also fig. 3. 


Curves of CO, excretion (solid black line) and O, absorption (dotted line) in milligrammes. 


Fic. 10.—Rat 3. See also figs. 4 and 5. 
Curves of CO, excretion (solid black line) and 0, absorption (dotted line) in milligrammes. 


76 


Carbohydrate Metabolism in relation to the Thyroid Gland 


MG.COz N.I N.2 N.3 TH.I TH.2 TH. 
500 


600 ; 


| 
i — ———— 
2483MG.COz 2789MG.COz 3030 MG.CO2 2951MG.COz 2865MG.COz 3019 MG.COz 


Fie, 11.—Rat 4. See also fig. 6. 


Curves of CO, excretion (solid black line) and O, absorption (dotted line) in milligrammes. 


EXPERIMENTAL OPERATIONS ON THE PITUITARY? 
By W. Buiarr BELL. (With fifty-seven illustrations in the text.) 


(Received for publication 22nd September 1916.) 


CONTENTS. 
INTRODUCTION ; a 
OPERATIVE TECHNIQUE 78 
THe EFFECTS OF THE OPERATIONS :— 
A. Control experiments . : 88 
B. Total extirpation of the pituitary ‘ 5 90 
C. Partial extirpation of the pituitary . 93 
(1) Anterior lobe removals . , , ; 93 
(a) Total removal of the pars anterior ; 93 
(b) Partial removal of the pars anterior : ; 96 
(2) Posterior lobe removals . : ; : 103 
(a) Total removal of the pars posterior ‘ Wie 3 103 
(b) Partial removal of the pars posterior } 104 
(3) Combined partial anterior and posterior lobe removals ; 108 
D. Clamping and separation of the stalk ; : 108 
E. Imitation tumours in the neighbourhood of the pituitary 116 
DiscussIoN oF RESULTS : ; ) ; ‘ s ; : 120 
CONCLUSIONS . , é : : ; 125 
INTRODUCTION. 


THE experimental work described in this communication was curtailed 
by the outbreak of war; and, for the same reason, the examination of the 
material obtained and the arrangement of it for publication have been 
greatly delayed and hampered. 

These investigations were carried out in order that the following points 
might be studied : 

(1) The general effects produced by pituitary lesions; (2) the effects of 
these lesions on the genitalia; (3) the effects produced by the pituitary 
lesions on the other organs of internal secretion. 

It was anticipated that evidence would be obtained in confirmation or 
refutation of the recent work on the same lines; while the possible acquisi- 
tion of fresh information was kept in view. 

Experimental operations on the pituitary, so frequently attempted since 
Victor Horsley (1886 (14)) first entered the field, have probably not been 
satisfactorily accomplished by as many as half-a-dozen operators. 


1 A Hunterian lecture was delivered by the author on these experiments at the Royal 
College of Surgeons, England, on 11th February 1916. 


78 Blair Bell 


I do not propose to give a complete historical survey of the subject, for 
it is hardly necessary again to describe the imperfect methods often 
employed and the unreliable results obtained, which have been so well dis- 
cussed by Crowe, Cushing and Homans (1910 (6)), Bied1 (1910 (4)), and 
others. It may, however, be said that although a large number of 
experimenters have attempted to operate on the pituitary—for the most 
part by the buccal route—in almost every case either the operation was a 
failure from a scientific point of view, or post-operative complications 
resulted in the death of the animal. This state of affairs obtained until 
Paulesco (1908 (16)), Professor of Physiology at Bucharest, with the aid of 
a surgical colleague, evolved the procedure, known as the bitemporal route, 
which has led not only to the accomplishment of successful operations, but 
also to more reliable results. Nevertheless, as already indicated, the 
published accounts of operations experimentally performed on the pituitary 
by the newer method of approach are still surprisingly few. It is probable 
that, apart from the investigations of Paulesco (1908 (16)), of Harvey 
Cushing and his colleagues (1909 (17)), (1910 (6)), and possibly also of 
Silbermark (1910 (18)), Biedl (1910 (4)), and Ascoli and Legnani 
(1912 (2)), no successful work has been done. Further, although Victor 
Horsley (1886 (14)) was certainly the first to perform extirpation 
experiments in this country, and probably in the world, hitherto there do 
not appear to have been any other attempts either by surgeons or 
physiologists in Great Britain—apart from later efforts (1911 (12)) by 
the operator just mentioned—to conduct investigations on these lines. 

The present researches, then, have been concerned in testing the correct- 
ness or otherwise of the experiments carried out by Paulesco, and by 
Cushing and his fellow-workers; and in attempting to gain further 
information concerning the experimental pathology of the pituitary. In 
all cases reliance as to the structural conditions present was placed only 
on the results of complete histological examinations. These histological 
observations included the examination of the pituitary tissues removed 
and of the site of removal; also of the genitalia, before and after opera- 
tion, and of the other endocrine! organs after the production of the 
pituitary lesions. Owing to the lack of laboratory assistance it was 
impossible to make detailed metabolic and clinical observations. 


OPERATIVE TECHNIQUE. 


In these experiments 27 bitches were subjected to operation, and of 
these two died as the result of the operative procedures, as distinct from 
the actual lesions produced on the pituitary. Only one of the deaths 
could be attributed to faulty surgical technique, and in this case the 
operation was abandoned owing to hemorrhage, from which the animal 


1 The term “endocrine” is used in this paper in aécordance with the usual custom in 
this Journal. 


Experimental Operations on the Pituitary 79 


succumbed shortly afterwards. ‘The other immediate death was due to 


an overdose of ether before the completion of the operation. Another 
animal died soon after total extirpation of the pituitary from some 
unknown cause—possibly from an overdose of the anwsthetic. All the 


other cases did well so far as the operative procedures were concerned. 

Excluding, then, the two bitches that died before the completion of 
the operation, 25 cases are left for consideration (Tables I-IX.). 

Most of the animals used were from four to seven months old, but 
a few were a little older. The younger the dog is, the easier the operation, 
owing to the thinness of the skull and lesser risk of serious intracranial] 
hemorrhage. 

Preliminary Procedures.—For ten days previously to the principal 
operation the animal received daily 10 grains of formamine in the food, in 
order that the cerebro-spinal fluid might be rendered antiseptic, as advised 
by Crowe (1909 (5)). 

All the animals should be weighed immediately before the first opera- 
tion, and at intervals subsequently. Owing to a misunderstanding on the 
part of the laboratory assistant, many of my animals were not weighed 
both before and after operation. 

Anesthesia was produced with ether by the “open” method a few days 
before the operation on the pituitary, and a small portion of one uterine 
horn together with part, or the whole, of the ovary on the same side were 
removed through a lateral abdominal incision for the purpose of control 
observations in connexion with subsequent changes in the genitalia. At 
the conclusion of this operation the whole of the top of the head and back 
of the neck was closely shaved, in order to lessen the time occupied at the 
second operation. In those cases in which a fatal result was anticipated— 
except in the case of dog No. 19—the removal of portions of the genitalia 
was not practised, but the preliminary shaving was always effected. 

Method of producing Anesthesia.—In view of the difficulty of 
working aseptically and comfortably during the operation on the pituitary 
in close proximity to the administrator, if the anesthetic were given in 
the ordinary way, I decided to use an intratracheal method for the 
administration of ether. This was found to be ideal after we had over- 
come the initial difficulties, which caused us to lose certainly one and 
possibly two out of the first three animals submitted to operation. 

In all the subsequent cases the anesthesia, conducted by iny laboratory 
assistant, Private Walter Plevin, was smooth, uninterrupted, and safe, and 
the animal was easily restored to consciousness as soon as the operation 
was completed by the administration of air alone through the intra- 
tracheal tube. 

In fig. 1 the apparatus used is illustrated. It is an easily made 
adaptation of the more complicated machines in general use for the intra- 
tracheal administration of ether to the human subject. Practice may be 
required in passing the soft rubber catheter into the trachea. The size of 


80 Blair Bell 


the catheter is determined by the diameter of the animal’s trachea, which 
the catheter should never fit closely. The insertion of the catheter was 
effected after the animal had been anesthetised with ether by the ordinary 
“open” method. » 

During the intratracheal administration of the anesthetic the animal 
rarely received pure ether vapour; it was usually sufficient, once the 
animal was fully anzsthetised, to continue the anzsthesia with a mixture 


Fic. 1.—Apparatus for the administration of intratracheal ether. (Photograph. ) 


A, tube from bellows; B, three-way tap indicator; C, tube to ether container; D, ether container; 
E, tube from ether container; F, tube to mercury pressure valve ; G, mercury pressure valve ; 
H, pressure manometer ; I, tube to catheter ; K, catheter. 


of air and ether vapour, regulated by means of a three-way tap to which 
an indicator was attached (fig. 1, B). By this means ether, ether and air 
mixed, or air alone could be pumped into the lungs under a uniform and 
limited pressure. 

The anesthetist sat at the side of the table opposite to the operator 
with his hand underneath the covering cloth and resting against the side 
of the animal, in order to judge of its condition: quiet, deep, and slow 
respiration indicated perfect anesthesia; rapid, shallow breathing too deep 
aneesthesia, while insufficient anzesthesia was shown by jerky and spasmodic 


Experimental Operations on the Pituitary S] 


respiration, or even by the animal coughing. ‘The heart-beat also gave an 
indication of the condition of the subject. 


Surgical Procedures. ~The animal, completely under the influence 


Fic, 2. —View of operation table showing also the sliding instrument table, the V-shaped 
block for animal’s head, and the tube leading to the ether apparatus. (Photograph. 


Fic. 3.—View of operation table and sliding instrument table covered with sterile cloths, 
as at the commencement of operation. (Photograph. ) 


of ether vapour administered intratracheally, was placed on its belly on an 

electrically heated table with the chin resting on a V-shaped depression 

cut out of a solid block of wood (fig. 2). The legs were fixed to the sides 

of the table, and the ears were tied together across the throat by means of 

a silk suture passed through the tips. Next, the eyes were carefully pro- 
VOL. XI., NO. 1.—1917. 6 


82 Blair Bell 


tected with dabs, while the previously shaven scalp was thoroughly purified 
with iodine dissolved in chloroform or spirit. The animal was then entirely 
covered with a sterilised sheet, in which there was a small aperture through 
which the operation was conducted (fig. 3). The special sliding instrument 
table attached to the operating-table (fig. 2) was also completely covered 
with a sterilised cloth (fig. 3). 

A long incision, extending backwards from the root of the nose in the 
mid-line over the vault of the skull as far as the occipital protuberance, 


Fic. 4.—View of a dog’s head, showing the line of incision. (Photograph x $.) 


was carried downwards and laterally behind the ear on the side from 
which the approach to the pituitary was to be made (fig. 4). This incision 
was found to be better than the Y-shaped incision which was adopted by 
Cushing and others, and which I, also, employed in the first few 
operations. 

Next, the skin over the right temporal region was turned down and 
the temporal muscle and pericranial fascia were carefully reflected from 
the underlying temporal bone, which was removed widely with a trephine 
and rongeur. The dura mater was opened with a triradiate incision, care 
being taken to avoid the vessels. The muscle flap was then loosely 


Experimental Operations on the Pituitary 83 


replaced together with the overlying skin, and attention was turned to 
the other side, from which the major portion of the operation was con- 
ducted: in my experiments this was always on the left side of the animal. 
The skin overlying the temporal region was raised on this side until the 
zygomatic arch was exposed. This structure was excised, together with 
the overlying aponeurosis, with a pair of bone-cutting forceps. Next, the 
temporal muscle was reflected as on the right side, and the temporal 
bone widely removed with a trephine and rongeur (fig. 5). On this side, 


Fic. 5.—View of the field of operation at the stage when the bilateral openings have 
been made in the skull. (Drawing. ) 


however, care was taken that the aperture made extended down as far as 
possible to the base of the skull. If there was any bleeding from the bone 
on either side it was readily stopped with bone-wax. 

In fig. 6 is seen a skiagram of the skull of one of the animals taken 
during life a few weeks after operation: the large aperture made on the 
left side is well shown. 

A head-light was now required; and the next steps of the operation 
were conducted by the operator single-handed, for it was necessary to 
manage the brain-retractor with one hand, while the manipulations in 
connexion with the pituitary were carried out with the other. It will be 


84 Blair Bell 


evident, therefore, that as no assistant is really required for the initial and 
later stages, these operations can easily be performed without help. But, 
in order to expedite matters, such as the cutting of ligatures, I was usually 
assisted by my then house-surgeon, Mr C. Baxter. 

The brain—that is, the temporal lobe on the left side—was carefully 
elevated with the special spoon-shaped retractor recommended by Cushing 
(fig. 7). As soon as the temporal fossa was crossed a thickened ridge of 
dura mater attached to the inner limit was seen. This marks the outer 
edge of the sella turcica, or rather the fossa to the edge of which the dura 


Fig. 6.—Radiograph of dog’s head taken during life some weeks after operation, 
showing the large aperture made in the left side of the skull. 


is attached, and of which the sella turcica is the most dependent part. 
The long, hooked knife (fig. 8) was then taken, and a slit made in the 
dura mater above the lower attachment just mentioned. Care was taken 
to carry the incision forwards rather than backwards in order to avoid 
wounding a large vessel frequently encountered in the dura mater. 
When the tip of the retractor was passed through the opening thus made 
a white glistening ridge or strand of reinforced dura was disclosed, and, 
when the beak was tilted upwards, the third nerve passing from behind 
forwards and above downwards was brought into view, and in front of 
this the carotid artery. Between these two structures, but further in, 
the pituitary body was exposed (fig. 9). This organ was easily recognised 


Experimental Operations on the Pituitary 85 


by its typical apricot colour, which is due to the extreme vascularity of the 
anterior lobe. 

All blood and cerebro-spinal fluid was now mopped out with small wool 
dabs held in forceps bent at an angle (fig. 10). As soon as the field was 
dry the rest of the operation planned was carried out. 


Fic. 7.—Soft metal spoon-shaped brain retractor. (Photograph x 3.) 


Most of my operations consisted in removing portions of the pituitary. 
I used for this purpose a special pair of aural forceps (fig. 11), which I 
found very convenient. It was impossible to obtain, or have made, an 


SS 


Fic. 8.—Long, hooked knife for incising the dura mater. (Photograph $.) 


instrument with spoon-beaks which were large enough to contain the 
pituitary body and which at the same time could be opened widely. With 
the instrument mentioned I was able to remove in several fragments most 
of the anterior lobe, and practically the whole of the posterior lobe intact. 
To remove the entire pituitary body I had a special instrument made: 
this consists of a lower blade terminating in a blunt hook in which the 
stalk of the pituitary is caught; the upper blade, which is formed like 


SO Blair Bell 


a chisel but is blunt, can be pushed in, so as to cut through the stalk 
caught in the hook (fig. 12). The lower attachments of the pituitary were 
always first separated with a fine blunt Watson-Cheyne dissector (fig. 13), 
before the stalk was cut through: after this the pituitary could be lifted 
out with a pair of bent forceps. This freeing of the lower attachments of 
the pituitary was found advisable in all operations except those in which 


Fic. 9,—View of the field of operation at the stage when the pituitary is first exposed. The 
circle of light from the head lamp is seen in the centre of the field. (Drawing.) 


the stalk was separated or clamped, or in which an artificial tumour 
was introduced. 

The operation was completed by the sewing of the temporal muscles in 
place, and the closing of the skin incision. 

Alarming hemorrhage sometimes occurred during the operation, 
especially in the older animals, but in all except one case this was 
controlled without difficulty; and probably there was a little careless- 
ness in the case that was lost, for one had got used to checking the 
hemorrhage easily with wool plugs, and had come to regard it as of little 
consequence. Nevertheless, the operation was found to be much less 
formidable than the previous descriptions of it led us to expect. Cushing 


Experimental Operations on the Pituitary 87 


states that after doing a large number of operations “it is not unusual for 
the operation to be completed within an hour after the beginning of 
anzesthesia.” On only one occasion—the first—did I exceed that time. 
The average time occupied by the actual operations was 39 minutes, 

The final determination of the character of the operation in the 
extirpation experiments was made in every case by a careful comparison 


Fic. 10,—Angled forceps for holding wool dabs with which the cerebro-spinal fluid and 
blood are mopped. (Photograph x 3.) 


Fic. 11.—Aural forceps for removing portions of the pituitary. (Photograph ~ }.) 


of the tissue removed at operation with the post-mortem histological 
findings. 

As it seems hardly worth while reduplicating the illustrations by 
reproducing more or less identical results, only those photographs and 
photomicrographs have been reproduced which illustrate most clearly 
the findings that may be considered typical and important. 

Post-operative Symptoms.—In no case were there severe complica- 
tions, such as serious sepsis or paralyses, as the result of the operative 
procedures; but sometimes there was an escape of cerebro-spinal fluid 
from the wound. Generally, the animals drank milk within a few hours 


88 Blair Bell 


of the operation, and seemed little affected the next day, when they 
ate meat, and were able to get out of their beds and walk about. 

Of the general symptoms following operations on the pituitary, 
polyuria and glycosuria are probably not infrequent — except after 
immediately lethal procedures, when there may be anuria—but as these 


Fic, 12.—Author’s chisel-hook for cutting through the stalk of the pituitary. (Photograph x 3.) 


phenomena may follow injury or stimulation of any part of the pituitary, 
I shall not discuss them at any length. 

With respect to the cachexia said by Cushing to be specific in con- 
nexion with certain pituitary lesions, I have been unable to verify his 


> a a 


Fic. 18.—Watson-Cheyne’s dissector. (Photograph x $.) 


conclusions, and am of the opinion that the supposed typical attitude 
attributed to the so-called “cachexia hypophyseopriva” (cf. fig. 46) is 
merely an attitude of weakness which is always seen in dogs in an 
advanced stage of emaciation and debility from any cause whatsoever. 

I shall discuss later the curious somnolence which may overtake the 
animals after some of these operations. 


EFFECTS OF THE OPERATIONS. 
Control Experiments. 


These were two in number, and in both cases the bitches were submitted 
to the same procedures as those adopted in the other experiments, even to 


Experimental Operations on the Pituitary 89 


the previous removal of a portion of the uterus and ovary. The pituitary 
body was exposed at the second operation, but no portion of it was re- 
moved ; instead, small pieces of tissue were excised from the base of the 


Fic, 144.— Dog 9 before control operation. (Photograph. ) 


Fic. 148.—Dog 9, 152 days after control operation. (Photograph. ) 


brain in the neighbourhood. Neither of these animals showed any 
symptoms until shortly before death, when one of them died with con- 
vulsions. This animal was probably poisoned, for another bitch which 
was chained up next to her died with convulsions at the same time. 
In both cases death occurred many months after operation, and in neither — 


90 Blair Bell 


were lesions found in the brain which could have accounted for the 
convulsive seizures. 

In Table I. are given the details of these control experiments. One 
of the bitches (No. 9) before and 152 days after operation is shown in 


TABLE I,—CoNnTrROL EXPERIMENTS. 


a = 
~ POM CIES. 5 rb) | 2 
2 foe hy bed = oT E P.M. findings. 
> oO =— > Los D = = 
S Ha, {S| q=) GLE 2 = Be | les 
Zs =I oo |e Setar rethers > | Ss aGuneere: ee 
g os os | Sees = aes || eS 
=z So WwW +s n Om DS ° Palos | = 2, 
© ee | 85 |88o08) ert Coteat en ye 
a AZ |sas6 3 as Se = | Macro- Micro- 
= OT tn eats tH — 2 n scopical. scopical. 
= a ans Se Sova 
a= a 


Dog 6! 9 mos. | Mar. 31) Sections Died 166 days None until | Nothing ab-} Pituitary 
show Sept. | last48hrs., normal normal 
small 13 of life, | 
piece of when fits) 
brain occurred 

9 | 94 mos. | Apr. 28} Sections | Killed (152 ,, | None Nothing ab-/| Pituitary 
show Sept. norma] normal 
small 27 


| 
piece of | 
brain | | 


' This animal and dog No, 2 were kept side by side in the animal house. Both died with 
convulsions within a few hours of one another many months after operation. Strychnine poisoning 
was suspected, but the examination of the stomach of this dog gave a negative result. 


figs. 144 and 148. It will be observed that there is no change in the 
animal except some slight increase in size corresponding with the 
increase in age. 


Total Extirpation of the Pituitary. 


This operation (figs. 15 and 16) was effectually carried out on six 
animals. In all cases a few cells of the reticulated portion of the pars 
intermedia must necessarily be left at the base of the brain, otherwise 
the third ventricle would be opened and part of the base of the brain 
removed. 

The first animal died shortly after the completion of the operation ; 
so soon that it is possible that death was due to an overdose of ether 
which was used too freely during the operation. Of the other five all 
_ died within a short time; that is to say, within periods ranging between 

22 and 36 hours. In all these five cases the animals recovered from the 
anesthetic, and were able to take nourishment freely. Before long, how- 
ever, they became somnolent, and although it was sometimes possible to 
rouse them from this condition, and to get them to stand and take food, 
they quickly became somnolent again as soon as they were left alone. 
After a few hours the respirations became very slow and coma set in; 
finally death supervened. The details of these operations are shown in 
Table II. 


Experimental Operations on the Pituitary 


Fic. 15.—The anterior and posterior lobes of the 
pituitary removed at operation from dog 1, 
(Photomicrograph x 15.) 


Fic. 16.—The base of the brain at the site of 
removal of the pituitary from dog 1. (Photo- 
micrograph x 15.) 


91 


o2Z Blair Bell 
TABLE II,—Torat or ALMOsT ToTAL REMOVAL OF THE PITUITARY. 
| oo & . 
2 | a Sap: ne oy E P.M. findings. = 
So cee =tunl etc See pen ovis =4 3 
avi S| Sie IN Basie sp] Se iligene agian = ae 
EH Riese ees 2628 SU Meech ee “ 
oO ee 85 Ss oecue| Salle saaes = s . = 
= yey Seo = Ze OF Macro- Micro- g 
= | ° Seas ia = = an scopical, scopical, | 
| | 
| Dog 1°9 mos. | Feb. 17 | Sections Died | Died a; Did not} No  he-| Stalk with | ? Anews- 
show total Feb. short | recover morrhage;| a few pars| thetic 
| anterior 17. | while | conscious-| small intermedia 
| and _pos- after ness blood-clot| cells at- 
| terior lobes opera- only tached 
| tion | 
5 Che », | Apr. 7] Sections Died | 24 hrs. | Dullness | Small Stalk with |Removal 
show total Apr. | and re-| blood-clot} a few pars | of pitu- 
anterior S| fusal of| in sella] intermedia “ itary 
and _pos- | food ; turcica cells and 
terior lobes | finally blood-clot 
| coma, attached 
| Respira- | 
| tions 10. | 
Pulse 140 
» 23 7 4, | Sept. 8] Sections Died! | 22 ,, Coma. Small Nothing but Removal 
show total | Sept. Respira- | blood-clot| smallblood-| of pitu- 
anterior 9 tions 13 | in track | clot the size | itary 
and _pos- of opera- | of pituitary 
terior lobes tion | 
5 27/3 4, | ,, 80] Sections Died 36 ,, | Dullness ; | Small A few pars Removal | 
| show total} Oct. | thencoma]} blood-clot} intermedia | of pitu- 
| anterior 2 in sella| cells with | itary 
| and — pos- turcica cysts below 
| terior lobes | 3rd_——sven- 
tricle, and 
a small 
| | | blood-clot 
» 29/33 ,, Nov. 9] Sections Died | 36 ,, Dullness | Small A few de- |Removal 
show total} Nov and re-| blood-clot} generated of pitu- 
| anterior 11 fusal of} in sella} pars inter-| itary 
and = pos- food ; turcica media cells 
| terior lobes finally lying in 
| coma blood-clot 
| | below 3rd 
| ventricle 
», 980) 4 | ,, 10} Sections Died | 36 ,, Coma Small A few pars |Removal 
| show pos-| Noy blood-clot| intermedia | of pitu- 
terior lobe| 12 in sella cells along | itary 
| | only turcica base of brain. 
| | | No sign of 
| | | anterior or 
| | | SS posterior 
| | lobe 


No observable changes occurred in the genitalia in the few hours of 
life subsequently to operation, nor were any definite changes found in the 
other endocrine organs in these circumstances. 
hyperplasia in the thyroid, for Cushing is very definite on this point, 
but in no case was any change to be discovered. A section of the thyroid 
of bitch No. 23 is shown in fig. 17. The organ is apparently quite normal. 


One anticipated finding 


Experimental Operations on the Pituitary 93 


Fic. 17.—Section of the thyroid of dog 23, 22 hours after removal of the pituitary. 
(Photomicrograph x 40.) 


Partial Extirpation of the Pituitary. 


Anterior Lobe Removals.—(a) Total Removal of the Pars 
Anterior.—In only two experiments was the anterior lobe almost 
completely removed (Table III.). It seems practically impossible to 


TABLE III.—ToraL or ALMost ToTAL REMOVAL OF THE ANTERIOR Loser. 


Com = ys 
= ee z oy t P.M. findings. cE, 
s ss | 2865S | 3 |f2.| sé 3 
La] o.oo ass oy ae aoe a = 
% Zz, 2s Sase gy | 2 os 22 hs 
ao 5 tO +s m C0 5 —'= S ao) °o 
o aS Zo SBom 42) ESS = 6 Macro- Micro- © 
= Aa 5250 2 ag te aga Macro Micro 2 
a 2 Be ae SI S a R scopical, scopical. = 
a | Be <) 
Dog 5 7 mos.}| Apr. 6| Sections Died |70 hrs. | April 7, | Nothing Blood-clotin |Removal 
show total Apr. none, abnormal] and around | of an- 
anterior 9 April 8, infundi- terior 
lobe none, bulum, As} lobe 
April 9, far as can 
extreme be seen the 
drowsi- pars an- 
NESS ; terior has 
| finally been Te- 
| coma moved en- 
tirely 
eS) OR) 5; » 9] Sections Died | 32 ,, Dullness | Nothing No pars an- [Removal 
show two) Apr. and re-| abnormal’ terior to be! of an- 
large pieces) 11 fusal of found. terior 
of anterior food ; | Poor section | lobe 
lobe thencoma | of region 


Pr a 


94 Blair Bell 


remove the entire anterior lobe without damaging the posterior. In 
fig. 18 are shown the portions of the pars anterior removed at opera- 


Fic. 18.—Section showing large portions of the pars anterior removed at operation 
from dog 5. (Photomicrograph x 15.) 


Fic. 19.—Section showing the base of the brain at the site of the pituitary 70 hours 
after operation in dog 5. (Photomicrograph x 15. ) 


tion from dog No. 5, and in fig, 19 is* seen the pituitary area at the 
base of the hae after operation. 


Experimental Operations on the Pituitary 95 


In both cases death followed the extensive removal of the pars anterior 
within a few hours. 


Fic. 20,—Section of the thyroid of dog 5, 70 hours after removal of a arge 
portion of pars anterior. (Photomicrograph * 40. ) 


Fic. 21.—Section showing large piece of pars anterior removed from dog 3. 
(Photomicrograph x 15.) 


Nothing abnormal was observed in the other endocrine organs after opera- 
tion. The thyroid of bitch No. 5, 70 hours subsequently, is shown in fig. 20. 


/ 


96 Blair Bell 


The genitalia, too, showed no changes in the short period of time that 
elapsed between the operation and the “death of the animals. 
(b) Partial Removal of the Pars Anterior.—lIt has been monianed 


Fie, 224,.—Dog 3, before operation. (Photograph. ) 


iD get) © Sat. \y Ee > 
aeosras ene ‘ 
f x. seas 


gad vege ope 
Att: 
hee 


Fic. 228.—Dog 3, 210 days after removal of a large piece of pars anterior. (Photograph.) 


that complete removal of the pars anterior alone is practically impossible, 
and that the removal of nearly all of it is usually fatal. Nevertheless it is 
quite easy safely to remove very large (fig. 21) or small portions of the 
anterior lobe; consequently, observations of the effects produced by these 
operations should be reliable. 


Experimental Operations on the Pituitary 


7 


In Table IV. are shown the results of partial removals of the pars 
There were five experiments, and in all the animals survived. 


anterior. 


é 


Approximate 


| 
| 
| 


| Dog 3 | 4 mos. 


», 24/43 ,, 


6 mos. 


TABLE 1V.—ParriaAL REMOVAL OF THE ANTERION LOBE. 


operation. 


| Mar. 10 Sections 


May 4) Sections 


| June 2 


July 5 | Sections 


Sept. 22 


| 


| 


— 
Z 43% sa 7 
2568 = 
=s—seE=+ = 
™%~ aoa eo od 
3 he & > 
zs o> z 
o2zseo oat 
ON = 
=>2e i. 

i Killed 
show large} Oct. 
portion of| 6 
anterior 
lobe 

Killed 
show fairly} July 


large piece} 3 
of anterior 


lobe 

Sections Killed 
show small} June 
portion of} 11 
anterior 

lobe 


show very 
large 
amount of 
anterior 


lobe 


Oct. 
aL 


Sections 
show very |. Nov. 
large 
amount of 
anterior 
lobe 


| 


Killed | 


Interval between 
operation and 


60 


death. 


Clinical 
symptoms. 


210 days} Mareh 11, 


none. 
March 12, 
drowsy. 
March 13, 


| very 


drowsy. 
March 14, 


| improved. 


Recovered 

Drank 
some milk 
1 hour 
after 
operation. 
Nosymp- 
toms. Re- 
| covered 


Ola. lions. Re- 


Killed 108 


40 


29 


” 


covered 


Drank 
some milk 
1 hour 
after 
operation. 
July 6, 
drowsy. 
July 7, 
drowsy. 
July 8, 
improved. 
Recovered) 

Animal 

| very weak) 

| through- 
out whole 


period 


| 


» = 
i i 2 
gS |S 
22/35 
"eo @ | OO 
oo | oe 
2olp 
= = 
arm. | grm., 
6680 5200 


4350 5350 Nothing ab- | Most of pars | 
anterior re- | 


P.M. findings. 


Macro- 
scopical, 


Uterus, 


ovaries, and 
breasts atro- 
phied. Thy- 


roid small 


Uterus, 
ovaries, and 
breasts 
slightly 
atrophied | 
| 
Nothing ab- 
normal. 
(Period too | 
short _ be- | 
tween opera-| 
tion and| 
death. ) 


normal : but 
no control | 
of genitalia 
taken at a) 
previous 
operation 


| Uter us, 
ovaries, and 
breasts very | 
slightly 
atrophied, 


| Thyroid 


very large 


plasia of ex- 
isting cells 


| of pars an- 


teriormixed 


with blood- | 


| clot 


moved. Pars 


Micro- 
scopical, 
Partial re- 
moval of 
pars ale 

terior 
| Partial  re- 
moval of 
parsanterior 
| 
Pituitary 
much  dis- 
turbed. 
Muchhyper- 


posteriorin- | 


tact 


| Most of the 


parsanterior 
removed. 

Pars pos- 
teriorintact 


It will be noticed that the results are not completely harmonious in regard 

to the details. 

As to the general effects: In no case was there any observable increase 
VOL. XI., NO. 1.—1917. ff 


98 Blair Bell 


in weight. Unfortunately only two bitches were weighed before as well 
as after operation, and of these No. 3—shown before, and 210 days after 
operation in figs. 224 and 22B—-lost weight subsequently to operation; the 
other, No. 19, increased in weight in accordance with its normal increase in 
growth. Some of the animals when recovering from the operation showed 
the peculiar condition of somnolence already described in connexion with 
total removal of the pituitary. As recovery occurred this state gradually 
passed off. 

Changes in the other endocrine organs were not found except, possibly, 
in the case of the thyroid from bitch No. 24. In this animal the thyroid 


Fic, 23.—Section of the thyroid of dog 24, 40 days after partial removal 
of the pars anterior. (Photomicrograph x 40.) 


was observed macroscopically to be considerably enlarged, but on histo- 
logical examination the organ was found to be normal (fig. 23). 

The variability in the results obtained would not be difficult to under- 
stand if it were only in the case of the removal of small portions of the 
pars anterior that no symptoms were produced, while excision of large 
portions produced changes in the general condition of the animal, in the 
genitalia and in the other endocrine organs. But these were not the results 
that were obtained; and it is difficult to understand why the removal of 
large portions from one animal—No. 19—should give rise to no ill effects, 
while the removal of smaller pieces, as in some of the other animals, should 
cause definite changes in the genitalia. 

In three out of the five cases in which portions of the pars anterior 
were removed the uterus (figs. 244 and 248) and ovaries (figs. 254 and 25B) 
were definitely atrophied. In these circumstances one finds that there is 


Experimental Operations on the Pituitary 99 


first of all atrophy in the muscular coats of the uterus, and that this is 
soon followed by atrophy in the endometrium. The changes in the ovaries 


Fig. 244.—Section of the uterus of dog 3 before operation. (Photomicrograph 40. ) 


Frc, 248.—Section of the uterus of dog 3, 210 days after partial removal 
of pars anterior. (Photomicrograph x 40.) 


are very striking: the whole ovary shrinks in size; and in detail one 
observes that the Graafian follicles degenerate—the ovum and epithelial 


100 Blair Bell 


contents (membrana granulosa) tend to disappear—the primordial ova 


Fic. 25a.—Section of the ovary of dog 3 before operation. (Photomicrograph x 40.) 


Fic, 258,—Section of the ovary of dog 3, 210 days after partial removal of 
pars anterior. (Photomicrograph x 40.) 


become opaque and lose their chromatin fibres, the interstitial cells vanish, 
and the stroma becomes fibrous. 


Experimental Operations on the Pituitary 101 


In two cases nothing abnormal was noted, but in one of these the length 
of time—nine days—that had elapsed between the operation and the post- 


Fic. 26,—Section showing large amount of pars anterior removed from dog 19. 
(Photomicrograph x 15.) 


Fic. 27.—Section showing normal uterus of dog 19, 108 days after extensive 
removal of pars anterior. (Photomicrograph x 40.) 


em examination was probably not sufficient for atrophic changes in 


mort 
In the remaining case no control was taken 


the genitalia to take place. 


102 Blair Bell 


before the operation on the pituitary, as it was intended that a fatal 
quantity should be removed. This, however, was not effected at the 


Fic. 28.—Section showing normal ovary of dog 19, 108 days after extensive 
removal of pars anterior. (Photomicrograph x 120.) 


Fic. 29,—Section showing posterior lobe removed at operation from dog 16. 
(Photomicrograph x 15.) 


operation, although a !arge amount was excised (fig. 26). The uterus and 
ovaries showed no changes from the normal 108 days later (figs. 27, 28). 


Experimental Operations on the Pituitary 


An examination of Table IV. seems to indicate that, in any case, a 


103 


con- 


siderable lapse of time must occur subsequently to operation if definite 


changes in the genitalia are to be expected. 


Fic. 30.—Section showing the site of the pituitary after removal of the pars 
posterior. It will be seen that a small portion of the pars nervosa at the 
neck was left behind. (Photomicrograph x 15.) 


Posterior Lobe Removals.—(a) Total Removal of the 


Pars 


Posterior.—In only one case was total removal of the posterior lobe 


effected (figs. 29 and 30). The details of this case are shown in Table V. 


The animal (No. 16)—shown before and after operation in figs. 314 
and 31B—had no symptom whatsoever. There was some increase in 
TABLE V,—Tovat REMOVAL OF THE POSTERIOR LOBE. 

om ne Gh = | ae | ae 
8 =e on a a hee ee P.M. findings. 
2 a S&S oe all ied = Prey thea [eee ate | en 
: B Ss "Eg os Dale ep ye Be a8 |29/¢3i— ~ 

o gS op io) a= ea m mes 26 as5 = 

BR * Se ee cS) oe a2 2 2 ae 

3 o Wt os 3 nwmas ans 2a =S3)s5 : 

o Sas as eGo eS S835] Ss | 708 | bec Macro- Micro- 
= co | 222 — | 83 eg Woda pie scopical. scopical. 
< aeags |g lee Shee ee eo Ee 

A= = 
oe — 3 [te es, LEC. Saree ey pee 
| grm. | grm. ; 

Dog 16'7 mos, June 1 | Sections Killed) 128days) None | 4700|5100|Nothing ab-|Shows _ab- 
show total Oct. normal. The] sence of the 
posterior 7 uterus, breasts,| pars —_pos- 
lobe and ovaries| terior, ex- 

had developed] cept the 
since the neck 
operation 


104 Blair Bell 


weight, but only in accordance with the growth of the animal. The 
uterus and ovaries continued to develop (figs. 832A and 32k, and 334A and 
338), and no changes were observed in the other endocrine organs. 


Fic. 31A.—Dog 16 before operation. (Photograph.) 


Fic. 318.—Dog 16, 128 days after removal of the pars posterior. (Photograph.) 


(b) Partial Removal of the Pars Posterior.—Of this experiment 
also there was only one case. This bitch died 199 days after operation, 
with convulsions (Table VI.). It has already been suggested that the animal 
may have been poisoned, for this and a control animal, mentioned above, 
both died within a few hours of one another with the same symptoms. 


Experimental Operations on the Pituitary 


Fic, 32A.—Section showing the uterus of dog 16, before operation. 
(Photomicrograph x 40.) 


Fic. 328.—Section showing the uterus of dog 16, 128 days after removal 
of the pars posterior. (Photomicrograph ~ 40. ) 


105 


Blair Bell 


106 


operation. 


dog 16 before 


y of 
(Photomicrograph x 120.) 


ar 


Section showing the ov 


HiGaiooAe 


Fic. 338.—Section showing the ovary of dog 16, 128 days after 


(Photomicrograph x 120. ) 


operation. 


| 
| 
| 


Case. 


The two bitches were chained side by side. 


Experimental Operations on the Pituitary 


brain to account for the convulsions. 


Approximate 
age, 


Date of 


Dog 2 7 mos, | Feb 


Case, 


Dog 17 6 mos, | June 25 Sectionsshow Killed 172 days| None 


107 


No lesion was found in the 


P.M. findings. 


Micro- 
scopical. 


a Saturday | 
night, and 
had been 


TasLe VI.—PartTiAL ReMovat oF THE Postrentor Lone. 
g°ea13|3z 2 : 
eiesca | a |e2.| ad [sale 
= | Sage|s |=88| 23 |23)/ 22 
= oat. e = | sez = > SE | <5 
eee fe he? | Of lk) gt | Mac. 
rs =| o >< ? a) 4 scopical, 
Aaees | 4 | eo ee as J 
; grm, 
. 24 Sections | Died |199 days, None until 10,000 11,200 Nothing Dog died on 
: showsmall | Sept. | last48-hours on abnormal 
amount of! 11 of life, when Apr. 
| posterior fits occurred 16 
| lobe which ulti- 


This animal and dog No. 6 were kept side by side in the animal house. 
vulsions within a few hours of one another many montlis after operation. 


mately 
caused death 


was suspected, but the examination of the stomach of dog No. 6 gave a negative result. 


TABLE VII.—PARTIAL REMOVAL 


Approximate 
age. 


} | 
| 
| 
| 


| 
| 


Date of 
operation, 


OF THE ANTERIOR AND POSTERIOR LOBEs. 


Bowes | eal Se . g ig 
meee | 3 bhs.| oe | es i<. 
eeea |; |2sa| 28 | 22 [Fs | 
eme5 | o | oes| £2 | Sf aa 
Beet) ailee: (OR f| Pe |e | ccopina 
See | ig. |e MS P< Na ae 
ey S | | 
a. 
| grm. {grm. 
| 7350 | 8550 Rather 
almost total Dec. | Animal fat 
posteriorlobe 14 | came on) 
and a small heat, and 
amount of had coitus | 
anterior lobe without | } 
| _ becoming 
| |_pregnant ee AL 
Sept. 15 Sectionsshow; Killed) 61 ,, None 5000 | 5500) Nothing 
almost total! Nov. | |(on Oct, | | abnormal) 
posterior S| | 1 after 
lobe with a opera- | 
medium tion) | 
amount of | 
anterior lobe 


dead some 
time before | 
post mortem 
could be 
made, Im- 
possible to 
cut good sec- 
tion of the 
parts —~ 


Both died with con- 
Strychnine poisoning 


P.M. findings. 


Micro- 
scopical. 


Sections 
show a large 
amount of 
anterior 
lobe and 
very little 

osterior 
obe 

Poor section 
of region 


So far as could be discovered, no changes in the genitalia or elsewhere 
had been caused by the partial removal of the pars posterior. 


108 Blair Bell 


Combined Partial Anterior and Posterior Lobe Removals.—In 
both the cases of this experiment (Table VII.) large portions of the pars 
posterior and small amounts of the pars anterior (fig. 34) were removed. 
In neither case were any symptoms or post-mortem appearances noted 
which could be ascribed to the operation. Both animals put on weight, 
bitch No. 17 becoming rather fat; this animal, moreover, came on heat 


Fig. 34.—Section showing portion of the pars posterior (on 
the right) and a small portion of the pars anterior (on 
the left) removed from dog 17, (Photomicrograph x 15. ) 


and had coitus, but did not become pregnant. The genitalia continued to 
develop, and no changes were noted in the other endocrine organs. 


Clamping and Separation of the Stalk. 


The details of these operations are shown in Table VIII. The results 
which they produce are probably identical, although it is possible that 
absolute severance of the stalk may produce more sudden and lasting 
effects than clamping. 

These experiments, as I shall point out more fully when discussing the 
results obtained by other workers, are of considerable interest, for in all 
three cases there was an increase in weight, and in two (Nos. 14 and 12) 
the condition of dystrophia adiposo-genitalis was produced. By no 
other operation was I able to obtain this result. 

In figs. 354 and 358 dog No. 14 is shown before and after operation, and 
in figs. 364 and 36B dog No. 12, before and after operation. In the second 
case, especially, an extreme condition of adiposity is to be seen: the body 


Experimental Operations on the Pituitary 109 


weight of this animal increased by 66 per cent. in 51 days. In fig. 37 
this bitch is shown laid open at the post-mortem examination.’ 


TaBLeE VIII.—CLAMPING AND SEPARATION OF STALK, 


Clamping the stalk. 


sz : = - 
5 i E: e ze = 25 4 2 2 5 P.M. findings. 
ry ug = Sere foe el) gs Ses > a2| Fs 
Ps | e?| 32 [g525 ¢ 523 82 (22 25 7 
= = (ey 15 F S = = 2 7 al 20 R ee Macro- Micro- 
| = ° eS Zi) S 7 ee scopical. scopical. 
grm. = grm. / 
Dog 14,7 mos. May 18 .. | Killed 129days Drank 6000 7100 Largeumount Sections show 
/ Sept. milk 1 (onJuly of sub- whole — pitu 
24 hourafter 28w.= cutaneous itary but cells 
operation. 7200) fut. Uterus, stain badly, 
| Fair ovaries,and | are separate: 
amountof breasts in-| from one an- 
adiposity, fantile. other in the 
| i.e. 20 °/, Thyroid ex- | pars anterior, 
increase in tremely and are 
weight in large | shranken 
71 days (? atrophied), 
The stalk is 
severed and re- 
placed by new 
fibrous tissue. 
Thyroid 
vesicles «lis- 
tended with 
/ colloid 
eee ae See 
Separation of the stalk. 
| 
| | | 
-Dog12 54mos. May 19 Killed 128 days For first 3/3000 5050 | Uterus, Sections show 
| Sept. | days ex- (same | ovaries,and  lineofcleavage 
| 24 | tremely | weight| breasts in-| belowpatch of, 
| / drowsy ; on Jul.| tenselyatro-| normal parsin-| 
_ afterwards 9) phied : termediacells. | 
| becameab-| very large | The rest of the 
| | | normally amount of) pituitary is) 
fat. In-) | subeutane- | embedded in| 
_ crease in ous fat fibrous tissue 
_ weight, | 
66 */, in 
) 5ldays | 
eee Os 55 \pAug-col| ~ ..; Killed 80 ,, | Increased 7400) 8100 Nothing ab- Poor section: 
Noy. | inweight | normal, ex-| tissues badly 
he | cept the fixed 
| breasts, | | 
| which are | 
infantile | 


The appearance of a dog with dystrophia adiposo-genitalis is 
remarkable, and no photograph does justice to the extraordinary degree 


1 This specimen is now in the Museum of the Royal College of Surgeons, England. 


110 Blair Bell 


of adiposity which may occur. In general appearance the animal 
becomes strikingly seal-like: the head and limbs look too small for the 
body, the fur becomes erect, and the breadth of the back causes it to 


Fic. 354.—Dog 14 before operation, (Photograph.) 


Fic. 35B.—Dog 14, 129 days after the clamping of the infundibular stalk. (Photograph. ) 


become flattened on the top. ‘The young animal may remain somatically 
infantile. 

Both of the animals which showed considerable increase in weight also 
showed complete atrophy of the genitalia (figs. 884 and 588, and 394 and 398) 
and mamma. Histological examination of the pituitary region showed 


Experimental Operations on the Pituitary 11] 


that at the line of separation and clamping there was a formation of new 
tibrous tissue, and that the cells of the underlying 


pars anterior were 
atrophied and widely separated (fig. 40). 


Fic. 864.—Dog 12 before operation. (Photograph. 


Fic. 36B,—-Dog 12, 51 days after the separation of the infundibular stalk. (Photograph. ) 


In disposition the animal is lethargic after recovering from the post- 
operative somnolence, which is pronounced. It sleeps a great deal, and 
when standing has a typical appearance: the tail and ears droop, and the 
animal appears to be only half-awake (figs. 358 and 368). In one case 
(No. 14) the thyriod was found to be very large indeed, and when 


112 Blair Bell 


Fic. 37.—Dog 12 laid open at the post mortem, 128 days after separation 
of the infundibular stalk. The enormous deposits of fat can be well 
seen; also the two horns of the atrophied uterus. (Photograph. ) 


Experimental Operations on the Pituitary 


Fic. 38a.—Section of the uterus of dog 12 before operation. 
(Photomicrograph x 40.) 


Fic. 388.—Section of the uterus of dog 12, 128 days after the separation 
of the infundibular stalk. (Photomicrograph x 40.) 


VOL. XI., NO. 1.—1917. 


113 


114 Blair Bell 


Fic, 394.—Section of the ovary of dog 12 before operation. 
(Photomicrograph x 120.) 


Fic. 392.—Section of the ovary of dog 12, 128 days after separation 
of the infundibular stalk. (Photomicrograph x 120.) 


Experimental Operations on the Pituitary 


Fic. 40.—Section at the site of the clamping of the infundibular 
stalk in dog 14, 129 days after operation, showing new fibrous 
tissue above and atrophic cells in the pars anterior below. 
(Photomicrograph x 120.) 


Fic. 41,—Section of the thyroid from dog 14, 129 days after the clamping 
of the infundibular stalk. (Photomicrograph x 40.) 


L15 


116 Blair Bell 


examined histologically the vesicles were seen to be enormously distended 
with colloid (fig. 41). 


Imitation Tumours in the Neighbourhood of the Pituitary. 


These experiments were three in number, but dog No. 26 was killed 
on recovering from the anesthetic, as she seemed to be in pain. The 
details of the experiments are shown in Table IX. The procedure was the 
same in all cases: the region of the sella turcica was exposed by the usual 
method, and the tumour was placed in proximity to the pituitary. 


TABLE IX.—IMITATION TUMOUR IN SELLA TURCICA. 


! 4 | 
— — 
. cS O o 5 . 
2g So Oo) cao : Wemeees ere P.M. findings. 
e ay fel AERIS om =e =, @ ie Sy 2 
=| oO ec Co: f=] 29 oe | ue) |e 
o a=] = Cys [Ss ay eo is) Qs eo 
n ma 2 or Osa oes So 2or 235 a o | 
Es} NST) Ys MD OO oS —'H (= TS —- 3s b= 
O a BF eh Gy ay no} Sie ee peo | Gra Macr Mier 
a | Alay | Sig ele nl ee eee ie eae acre | acer 
ae | ° SSS Ge i | & By aD [2 Shs scopical, scopical. 
Aan 8 —m | Be | = = | 
| pas |) | | | 
nee | 
| ; | erm. | grm. 
Dog 20'6 mos. July 6) aa Killed 98 days |Greatemacia- ... .... Largetumour Large cystin 
Sept. tion. Glyco- at edge of parsanterior 
| 27 | suria sella turcica, 
| Uterus, ov- | 
aries and_ 
| breasts at- 
rophied | 
Ie Zilee se Sepia 7 sae | Killed} 57 = ,, Well devel-| 8200|9800| Tumour in| The lobesare 
Nov. oped and front of pitui-| displaced in 
3 fat. Nearly tary. Noth-| their  rela- 
. | . . 
| 20% increase ingabnormal) tion to one 
| in 57 days | another 
» 26/45 ,, | Sept. 29 bide Killed|1 hour |Seemed in ... ... | Large tumour 
| Sept. pain, and occupying 
29 | therefore sella turcica | 
killed | 
| 


The artificial tumour was made of wax mixed with barium oxychloride, 
and was sterilised by heat. This substance when softened was easily 
placed in situ after being moulded into the shape of a bun (figs. 42 
and 43). The X-ray photographs were taken during life some weeks after 
operation. It will be observed that in neither case does the artificial 
tumour occupy the site of the sella turcica centrally, but there is no doubt 
from the post-mortem examination that in both cases the pituitaries were 
somewhat displaced (fig. 44), and in one case (No. 20) the pars anterior 
contained a large cyst (fig. 45). 

In one animal (No. 20) there was very considerable emaciation, 
with slight glycosuria. With regard to the general condition of this 
animal (fig. 46), it will be noticed that it presents the appearances 
described by Crowe, Cushing and Homans (1910 (6)), and stated to be 
due to a specific “cachexia hypophyseopriva.” It has already been 
mentioned that in my experiments no evidence was obtained of any such 


Experimental Operations on the Pituitary 


AN 1 
a WA 


Fic. 42. Radiograph, taken during life, of artificial tumour in dog 


99) 


118 


Blair Bell 


Fic. 43.—Radiograph, taken during life, of artificial tumour in dog 


20. 


Experimental Operations on the Pituitary 119 


Fic, 44.—Section of the pituitary from dog 22, 57 days after operation, 
showing the displacement of the pars anterior and pars posterior 


caused by an imitation tumour, (Photomicrograph x 15.) 


_— 


Fic. 45.—Section of the pituitary of dog 20, 98 days after operation, 
showing a cyst in the pars anterior caused by an imitation tumour. 


(Photomicrograph x 15.) 


120 Blair Bell 


specific condition. It is typical in the dog of general emaciation and 
weakness, due to any cause. This animal had some slight suppuration in 
the abdominal wound, and also contracted mange; consequently it is 
difficult to know how much of the emaciation to ascribe to pituitary 
irritation. 

The other animal (No. 22) showed no symptom whatsoever. ‘There 


Fic, 46.—Dog 20, 98 days after operation, showing the emaciation and attitude 
of weakness caused by glycosuria due to the pressure on the pituitary of 
an imitation tumour. (Photograph. ) 


was a considerable increase in weight, which was probably due to rapid 
growth and not to obesity. 


DISCUSSION OF RESULTS. 


It will now be of interest to see how far the foregoing experiments 
confirm or contradict the work of others. In this connexion it will be 
sufficient to consider the pioneer work of Paulesco (1908 (16)), and the 
subsequent experiments of Cushing and his colleagues (1909 (17)) and 
(1910 (6)) and of Bied1 and his associates (1910 (4)). 

The work of Aschner (1910 (1)) is less reliable, for although this 
investigator was able to produce certain of the abnormal phenomena that 
had been previously recognised by others, his methods, which have been 
justly criticised by Biedl, were not exact, since he used the oral route. 
Ascoli and Legnani (1912 (2)) also, so far as I can gather, do not appear 
to have clearly recognised that different lesions produce different results. 

Paulesco’s work, on the other hand, is of the highest merit, for by 
introducing the bitemporal route he at once placed the experimental 
possibilities on a sound basis. The results which he obtained may be 
summarised as follows :— 


Experimental Operations on the Pituitary 121 


. Complete extirpation of the pituitary caused death in a short time. 

2. Partial removal of the pars anterior caused no symptom other than 
adiposity. 

3. Extensive or complete destruction (thermo-cautery) of the pars 
anterior resulted in death. 

4. Removal of the pars posterior caused no symptom. 

5. Separation of the stalk resulted in the death of the animal. 

6. Separation of the pituitary from its bed in the sella turcica pro- 

duced no symptom. 


Cushing and his fellow-workers, as the result of two series of careful 
experiments, in which they adopted the technique introduced by Paulesco 
with slight improvements, obtained results very similar to his. Indeed, 
the only differences noted were in regard to partial removal of the pars 
anterior and to separation of the stalk. 

Cushing and his colleagues found that separation of the stalk produced 
the same effects as total removal with immediate transplantation. They 
also believed that the adiposity which occurred in their animals after partial] 
removal of the pars anterior was specific; that is to say, while Paulesco 
had observed that the animals might become fat, Cushing and his col- 
leagues were the first to recognise the importance of this adiposity, and 
to note that it was identical with the pathological condition previously 
known as dystrophia adiposo-genitalis, since there was also genital 
atrophy. Further, these investigators found that in young animals per- 
sistent infantilism occurred after partial removal of the pars anterior.' 

Cushing also made the striking discovery in regard to this condition 
that the subnormal temperature always found with dystrophia adiposo- 
genitalis can be raised by injections of an extract made from the pars 
anterior. This he called the “thermic reaction.” On the other hand, 
according to the same authority (1912 (8)), the low blood-pressure and 
carbohydrate tolerance associated with this syndrone are relieved by 
injections of infundibulin (posterior lobe extract). 

Again, Cushing and his fellow-workers found that although total 
extirpation was a fatal operation, the effect was not so rapid in young as 
in older dogs. 

Lastly, Cushing described a condition of “cachexia hypophyseopriva ” 
which was considered specific of deprivation (complete or almost complete) 
of pars anterior secretion.. 

Bied1 (1910 (1)), without giving details, states that he has confirmed all 
Cushing’s findings, except in regard to stalk separation, which operation, 
in agreement with Paulesco, he found to cause death. It is hardly worth 
while to dwell on Bied1’s results in the absence of details other than those 
given in his book (1910 (4)). 

1 Aschner also claims to have obtained dystrophia adiposo-genitalis by partial 


removal of the pituitary by the oral route. There appears to be no doubt, however, 
that Cushing made the first communication on the subject (1909 (7)). 


122 Blair Bell 


Silbermark (1910 (18)), in the reference given by Bied] (with whom 
he worked), discusses the technique of the operation. Apparently the 
results he obtained with Bied] are only recorded, without details, in 
Biedl’s work. 

The results of my experiments do not entirely confirm the work of 
Cushing and his associates, which is undoubtedly the most reliable and 
satisfactory of all the experimental work carried out on the subject. It 
will therefore be of interest to discuss the points of confirmation and con- 
tradiction, and to find, if possible, some explanation of the differences. 
Although I am far from convinced by the evidence of the small series 
of operations recorded here that the results obtained will stand the test 
of further research, nevertheless they appear to fit in with the most reason- 
able explanation of pituitary activity, as I shall explain directly. 

The results of my experiments concerning the effects of total extirpation 
of the pituitary, and of the removal of very large portions of the pars 
anterior, confirm the statements of Paulesco and Cushing that such 
procedures are fatal. 

Sweet and Allen (1913 (19)) alone of recent investigators deny that 
the pituitary is essential to life; but it appears to me that their technique 
is open to criticism. 

My experiments also confirm the fact demonstrated by Paulesco and 
Cushing that the removal of the pars posterior produces no symptom. 
Further, I have been able, by means of the control specimens removed 
before the operation on the pituitary, to show that the genitalia not only 
do not undergo atrophy, but continue to develop in the young female after 
removal of this portion of the pituitary. 

With regard to the points wherein my experiments produced results 
ditferent from those obtained by Paulesco, Cushing, and others, the most 
striking is undoubtedly in connexion with the production of dystrophia 
adiposo-genitalis. Whereas Cushing—and probably Paulesco,although 
he failed to recognise the importance of the condition—found that partial 
removal of the pars anterior was the lesion responsible for this syndrome, 
in none of the cases in which I removed portions of the pars anterior did 
dystrophia adiposo-genitalis supervene, although when sufficient was 
removed, and there was a considerable lapse of time between the operation 
and death, genital atrophy was usually found. In one case there was an 
actual loss of weight in a young animal in 210 days. This animal remained 
stunted. In other cases the animals increased in size. 

I found, however, that the syndrome dystrophia adiposo-genitalis 
followed clamping and separation of the infundibular stalk. In two out 
of three cases there was atrophy of the genitalia, with considerable adi- 
posity ; in one case the increase amounted to 66 per cent. of the body weight 
in 51 days. 

It is not impossible to reconcile these diverse findings, especially if we 
study the difficulties Cushing encountered when he attempted to make 


Experimental Operations on the Pituitary 123 


his experimental results conform to his clinica] observations. Believing 
that reconciliation was not possible, he was tempted to throw over his 
experimental results in favour of the clinical evidence that was in conflict 
with them. 

It will, I think, be sufficient to call attention to the chief perplexity 
which Cushing was called upon to face. As we have seen, the results 
of his experimental work indicate that dystrophia adiposo-genitalis 
is due to anterior lobe insufficiency. But in his clinical experience Cushing 
found, as already mentioned, that the only symptom of the syndrome 
dystrophia adiposo-genitalis relieved by anterior lobe extract was 
the subnormal temperature. While, on the other hand, the low blood- 
pressure, and the carbohydrate tolerance—and, as far as I can understand 
from his later writings, the genital dystrophy—were mitigated by posterior 
lobe extract. In view, then, of these clinical observations, how was it 
possible to attribute this syndrome to the experimental removal of portions 
of the pars anterior, as Cushing himself and others had done’ Cushing 
solved the question by rejecting his experimental results. 

The results I have obtained after clamping and separation of the stalk 
appear to explain the paradoxes. Such an operation could only interfere 
with the blood-supply of the whole organ; and, if the infundibulin does 
pass directly into the third ventricle, stop this source of supply. 

It is, however, hardly possible that the pars posterior and its secretion 
has anything to do with the matter, for all recent investigators are agreed 
that the posterior lobe can be removed without producing any symptom 
whatsoever. Further, since I found it possible to remove large portions of 
the pars anterior and the entire pars posterior without causing dystrophia 
adiposo-genitalis, but was able to produce this syndrome by clamping 
and separating the stalk, it is obvious that interference with the blood- 
supply to the pituitary produces the condition. There seems little reason 
to doubt, then, that this syndrome is primarily produced by insufficiency 
of the pars anterior; but it appears certain that the only sure way to etiect 
this is to interfere with the blood-supply. If this is done we find the 
cells of the pars anterior become shrunken, atrophic, and discrete—a 
state of affairs which is always found in the human subject afflicted 
with dystrophia adiposo-genitalis. 

It is now necessary to consider how the foregoing statements can be 
reconciled with the facts that removal of the posterior lobe causes no 
symptoms, yet infundibulin relieves some of the symptoms—the lowered 
blood-pressure and the carbohydrate tolerance—in dystrophia adiposo- 
genitalis. 

I have long held (1913 (3)) that to explain these facts we must look upon 
the pituitary as one organ and not two. Further, from the clinical and 
experimental evidence of this syndrome, and from other evidence which I 
need not detail here, it is probable that the view of Herring (1908 (13)) 
concerning the determination of the secretion of the pars posterior solely 


124 Blair Bell 


and directly into the third ventricle cannot be sustained, and that this 
secretion, if required, can be taken up, like other internal secretions, by 
the blood stream. It is to be remembered that the secretory cells of the 
posterior lobe —the cells of the pars intermedia —are derived from the 
same source as those of the pars anterior; consequently, while clamping 
and separating the stalk interferes with the blood-supply to all these 
cells, the removal of the pars posterior does not remove those situated 
at the base of the brain, nor does such an operation interfere with 
the pars anterior. Hence it is that it becomes necessary to look upon 
the functions of the pituitary as a whole, and to consider this structure as 
one organ and not two. The fortuitous juxtaposition of the epithelial 
cells and the pars nervosa has probably no relation to the vital—essential 
and beneficial—functions with which the pituitary is concerned. Even 
if secretion from the pars nervosa does pass into the cerebro-spinal 
fluid, there is not the shghtest evidence to show that this is essential, 
beneticial, or even the normal method by which infundibulin is taken 
up by the animal economy. 

Special attention has been directed by Cushing to the peculiar 
somnolent condition in which the animal may exist for some time after 
operations which decrease the pituitary secretion, especially that of the 
pars anterior. This state, which has already been described, is quite 
characteristic. It may exist in different degrees from a deeply comatose 
condition to merely mental lethargy. If the animal becomes really 
comatose, as is the case after complete and almost complete pituitary 
extirpation, death always, in my experience, supervenes. But some 
animals—for example, dog 3 in my series—become somnolent for many 
days, and must be roused and lifted out of their beds in order to get them 
to take food. This they readily do as soon as they are sufficiently aroused. 
Animals that recover usually pass from this condition into one of mental 
lethargy, which either disappears in time or persists—according to the 
permanence or otherwise of the diminished secretion. 

Cushing and Goetsch (1915 (10)) have likened this condition to that 
of hibernation, a state which Gemelli (1906 (11)) first suggested might be 
due to functional hypopituitarism. And it is interesting to recall the fact 
that in hibernation one sees exactly the same histological picture—shrunken, 
inactive cells—that one observes in the experimental cases in which lethargy 
persists, and in dystrophia adiposo-genitalis in the human subject. 

There is little to be said at present concerning the relation of pituitary 
lesions to polyuria and glycosuria. Cushing and his colleagues are now 
engaged in investigating these questions (9), and their work seems likely to 
revolutionize many existing opinions. Meanwhile, no good purpose would 
be served by a rediscussion of the present views. 

With regard to the experiments in which artificial tumours were placed 
in the neighbourhood of the pituitary, my experiments are too few to 
enable me to do more than conclude that neighbourhood tumours may 


Experimental Operations on the Pituitary 125 


cause irritation with glycosuria and wasting, while tumours interfering 
with the stalk may produce carbohydrate tolerance. In the human subject 
it is, of course, well known that tumours in the neighbourhood of the 
pituitary usually lead to the syndrome dystrophia adiposo-genitalis, 
by causing atrophic changes in the secretory cells. 

Paulesco (1911 (15)) has published a paper bearing on the experimental 
aspect of this subject, but I have been unable to refer to it, for at present 
it is unobtainable in this country. 


CONCLUSIONS. 


1. The pituitary body is an organ that is essential to life: its removal 
causes death within a few hours. In the cases which survive for longer 
periods the removal has probably not been complete. 

2. The removal of very large portions of the pars anterior is incom- 
patible with life. It appears certain from the evidence at our disposal that 
it is the loss of this portion of the organ which proves fatal when total 
extirpation of the pituitary is practised. 

3. Partial removal of the pars anterior may, if sufficient quantity be 
removed, cause genital atrophy. This may occur in the absence of any 
other symptom, althongh the animal may also remain undersized. 

4. Neither partial nor complete removal of the pars posterior causes 
any symptom. The genital organs remain normal after operation, and 
young animals continue to develop. Hence the secretion of the pars 
nervosa is neither necessarily beneficial nor essential to life. 

5. Partial removal of the partes anterior and posterior causes no symp- 
tom provided only a small portion of the pars anterior be removed. 

6. Clamping and separation of the infundibular stalk, by interfering 
with the blood-supply and so causing degeneration in the cells of the partes 
anterior and intermedia, lead to the condition known as dystrophia 
adiposo-genitalis. 

7. Artificial tumours in the neighbourhood of the sella turcica may pro- 
duce irritation, which is accompanied by glycosuria and emaciation : or by 
interfering with the blood-supply may lead to degenerative changes in the 
cells of the pars anterior, and so give rise to the syndrome dystrophia 
adiposo-genitalis. . 

8. The pituitary body appears to be one organ and not two; and the 
essential and beneficial secretion is taken up by the blood stream, as in the 
case of the other organs of internal secretion. 


The investigations described in this paper were carried out in the 
Pathological Department of the University of Liverpool, and I am indebted 
to Professor Ernest Glynn for the facilities atforded. The expenses were 
defrayed out of a fund placed by Mr J. Arthur Smith at my disposal for 
scientific research. 


XX1. 


Experimental Operations on the Pituitary 


REFERENCES. 
(1) Ascuyer, B., Wien. klin. Wochenschr., 1910, xxx. D7 2: 
(2) Ascot, G., and T. Leanant, Miinch. med. Wochenschr., 1912, lix. 518. 
(3) Bett, W. Buarr, Arris and Gale Lectures, R.C.S., Teenie 1913, 1. 809, 937. 
(4) Brevi, A., Innere Sekretion, 1910. 
(5) Crows, S, J., Johns Hopk. Hosp. Bull., 1909, xx. 102. 
(6) Crows, S. J., H. Cusaine, and J. Homans, Johns Hopk. Hosp. Bull, 1910, 
127. 
(7) Cusuine, H., Communication International Congress, Budapest, 1909. 
(8) Cusine, H., The pituitary body and its disorders, 1912. 


(9) Cusuine, H., Private communication. 

(10) Cusine, H., and E. Gorrscu, Journ. Exper. Med., 1915, xxii. 25. 

(11) Gemexu, A., Arch. p. le sci. med., 1906, xxx. 341. 

(12) HanDELsMaNN (no initial given in original) and V. Horstey, Brit. Med. 


Journ., 1911, 11. 1150. 


(13) Herrine, P. T., Quart. Journ. Exper. Physiol., 1908, i. 12i. 

(14) Horsey, V., Lancet, 1886, 1. 5. 

(15) Pautesco, N. C., Ann. de Biol., 1911, i. 221. 

(16) Pautesco, N. C., L’hypophyse du cerveau, Paris, 1908. 

(17) Reprorp, L. L., and H. Cusnine, Johns Hopk. Hosp. Bull., 1909, xx. 105. 
(18) Siupermark, M., Wien. klin. Wochenschr., 1910, xxi. 467. 

(19) Sweet, J. E., and A. R. Auten, Ann. Surg., 1913, lvu. 485. 


THE CHEMISTRY OF FOSSIL BONE. By J. ArGcyLti CAMPBELL, 
Singapore. (From the Departments of Physiology of the Uni- 
versity of Edinburgh and of the Medical College, Singapore.) 


(Received for publication 31st August 1916.) 


THIs research, suggested by Sir Edward Schifer, from whom the material 
was received, gives the results of analysis of three specimens of fossil bones 
—one human, one extinct marsupial, and one pinniped. 

The first bone, a right humerus from a prehistoric man, was received in 
January 1915. It was found near North Berwick, and was well preserved 
and entire, its brownish-red colour being due probably to the soil in which 
the bone had been embedded. 

The second bone, a portion of a rib of Diprotodon, found at Kings- 
thorpe, Darling Downs, Queensland, was received in September 1914. This 
specimen appeared petrified throughout. 

The third specimen consisted of two fossil seal bones, a right scapula 
and a left astragalus, from a young animal. They were taken from the 
Portobello clay by Dr Gordon, Professor of Geology, King’s College, 
University of London, and received by me in January 1916, well preserved 
and entire, possessing in some degree the moist, shining appearance and the 
great toughness of recently cleaned bones. 

Since geological measurements of time are not definite, nothing can be 
said regarding the age of these bones beyond the fact that they are prob- 
ably all many thousands of years old. The humerus belongs to the 
Prehistoric or Post-glacial epoch, and the specimens of seal bones must be 
referred to the same period, the clay in which they occurred belonging to 
the age of the 100-feet beach.!| Diprotodon was a large marsupial, attaining 
the bulk of a rhinoceros or hippopotamus, and inhabiting Australia during 
the Pleistocene period. 

The analysis of the humerus was carried out in the Physiology Depart- 
ment, University of Edinburgh, the other specimens being analysed in 
the Government Analyst’s Department, Singapore. ‘Two different portions, 
A and B, of the shaft of the humerus were examined separately, A being 
taken from the middle of the bone, B from nearer one extremity. The rib 
bone was also divided into two portions, A and B. 

1 Geological Survey, Memoirs of the Edinburgh District, p. 335. This information was 
supplied by Professor Gordon. 

VOL. XI., NO. 2.—1917. 9 


128 Argyll Campbell 


The bones were scraped clean and then powdered as finely as possible. 
The seal bones were very tough compared with the others. 

Volumetric methods were employed in the estimation of calcium, 
magnesium, and phosphorus in the humerus, and in portion A of the rib; 
with these exceptions gravimetric methods were employed for analyses of 
the ash. 

The following is a short summary of the method used to estimate each 
constituent. 

WATER. 


The powdered bone was dried at 110° C. This caused a loss of some of 
the water only. During incineration (see below) the bone lost more of its 
weight than that accounted for as protein, fat, and water driven off at 
110° C. As will be seen in the tables, I have assumed this loss to be 
water driven off at a temperature higher than 110° C. 


ORGANIC SUBSTANCES. 


Protein.—For qualitative examination the salts were first removed by 
dilute hydrochloric acid and the residue analysed for mucin and collagen. 
For quantitative estimation the total nitrogen was obtained by Kjeldahl’s 
method. The average of Hoppe Seyler’s percentages of nitrogen in all 
proteins (16 per cent.) was used in calculating the amount of protein in 
the bone. 

Fat was estimated by extraction with ether (Soxhlet’s method). 


INORGANIC SUBSTANCES. 


Total Ash was estimated by incineration in a platinum crucible, with 
subsequent addition of ammonium carbonate to recarbonate the lime. 

Chlorine, Sulphur, and Fluorine.—Only qualitative tests were 
made. 

The following volumetric methods were employed :— 

Calcium.—The powdered bone was submitted to combustion by acid 
incineration. The calcium was precipitated as calcium oxalate. dissolved 
in acid, and titrated against potassium permanganate. 

Phosphorus Pentoxide.—After incineration the phosphorus was pre- 
cipitated by ammonium molybdate. The precipitate was dissolved in excess 
of N/2 sodium hydroxide, and the excess estimated by titration against 
N/2 sulphuric acid. 

Magnesium.—After acid incineration and removal of the calcium as 
calcium oxalate, the magnesium was precipitated as ammonium-magnesium- 
phosphate. The precipitate was dissolved in acetic acid and the solution 
titrated against uranium nitrate. The magnesium was estimated from the 
amount of phosphate present. 

The following gravimetric methods were employed :— 

. Carbon Dioxide.—The carbonic acid was liberated by dilute hydro- 


The Chemistry of Fossil Bone 129 


ehloric acid, and after having been purified from moisture by sulphuric 
acid and from hydrochloric acid by copper sulphate, was absorbed in a 
weighed soda-lime tube. 

Silica was separated, and estimated in the usual manner by evapora- 
tion to dryness with hydrochloric acid. 

Trimanganic Tetroxide.—Ferric chloride was added to the filtrate 
from the silica; and the phosphate, alumina, and excess of iron were 
removed by the basic acetate method. Manganese was estimated in the 
filtrate as trimanganic tetroxide, after precipitation with bromine and 
ammonia and ignition. 

Calcium Oxide.—tThe tiltrate from the manganese hydrate was boiled 
and the calcium precipitated by ammonium oxalate and weighed as 
calcium oxide. 

Magnesia.—The ammonium salts were removed from the filtrate from 
the calcium oxalate by evaporation to dryness with nitric acid. The 
residue was dissolved in a few cubic centimetres of hydrochloric acid, the 
liquid made slightly alkaline with ammonia, and filtered. Magnesia was 
estimated in the filtrate as pyrophosphate in the usual manner. 

Phosphorus Pentoxide.—Half a gram of ash was taken, and after 
the removal of silica the phosphoric acid was precipitated by ammonium 
molybdate in nitric acid solution. The ammonium phospho-molybdate 
was dissolved in ainmonia and the phosphorus precipitated by magnesia 
mixture, ignited, and weighed as pyrophosphate. 

Ferric Oxide and Alumina.—tThe silica was removed trom 2 grm. 
of the ash. The filtrate was nearly neutralised by ammonia, and the 
phosphates of iron, aluminium, and some calcium were precipitated by 
ammonium acetate, ignited, and weighed (Fe,O,, Al,O,, CaO, P,O;). The 
resulting material was dissolved in hydrochloric acid, neutralised, and the 
calcium removed by precipitation as oxalate in a solution made freely acid 
with citric acid. This precipitate was ignited and weighed as oxide. The 
filtrate from it was made slightly alkaline with ammonia, and the phos- 
phorus precipitated by magnesia mixture. This gave the phosphorus 
pentoxide. ‘The iron was precipitated by passing sulphuretted hydrogen 
gas through the solution. This precipitate was dissolved by hydrochloric 
acid, oxidised with nitric acid, and precipitated as ferric oxide with 
ammonia. This was filtered off, ignited, and weighed. Alumina was 
estimated by difference. 


[ TABLE. 


130 


Ash 


Fat 
Ossein 
Water 
Water ? 


Ash 


Fat 
Ossein 
Water 
Water ? 


Fat 
Ossein 
Water 


Argyll Campbell 


RESULTS 


OF ANALYSES. 


Humerus ot Prehistoric Man. 


hoastO) = 


2°65 
5°47 
6-96 
9°22 


100-00 


76:43 


2°02 
4°91 
6°54 
10°10 


100-00 


Rib Bone of Diprotodon. 


82°00 < 


0-00 
0:00 


A. 


(110° C.) 
(above 110° C.) 


(110° C.) 
(above 110° C.) 


A. 


26°73 
35°40 
40 
5°50 
+ve 
+ve 
+ve 
+ve 


Organic = 812 
Inorganic = 91°88 


Organic = 6°93 
Inorganic = 93:07 


13°87 
50°90 
10°80 
3°18 
90 
trace 
trace 
+ve 
+ve 
—ve 


18°00 (loss during incineration) 


100°00 


Organic = 000 
Inorganic = 100:00 


Ash 


Fat 
Ossei1 
Water 


Ash 


Fat 
Ossein 
Water 
Water ? 


Ash 


Fat 
Ossein 
Water 
Water ? 


The Chemistry of Fossil Bone 131 


b. 
, P.O, 14-22 
CaO 47°39 
CO, 13°80 
Si0, 4°05 
Fe,O 2°24 
84:18 / Al,O 1°42 
Mg trace 
Mn,({ ) ‘HR 
SO, +ve 
Cl + Ve 
Fl ve 
0-00 
0°00 
15°82 (loss during incineration) 
100:00 Organic 0°00 
Inorganic = 100°00 
Scapula of Seal. 
P.O, 26°26 
CaO 42°75 
SiO, 0°32 
MgO trace 
69°69 ~ Fe,0, trace 
CO, + ve 
SO,» awh h= 
Cl +ve 
Fl -ve 
263 
18°71 
3°80 (110° C.) 
5°17 (above 110° C.) 
100-00 Organic =21°34 


Astragalus of Seal. 


P.O; 
CaO 
SiO, 
MgO 
62°16 . FeO, 
CO, 
SO, 
Cl 
Fl 
2°86 
26°40 
400 (110°C.)  ~ 
4°58 (above 110° C.) 
100°00 


Inorganic = 78°66 


23°71 
33°02 
0°57 
trace 
+ve 
+ve 
+ve 
+ve 
— ve 


Organic =29°26 
Inorganic = 70°74 


N.B.—There was not sufficient material t6 complete the analyses of the ash in the 


seal bones. 


132 Argyll Campbell 


ZALESKY’S ANALYSES OF DRIED MACERATED Bones. 


' cal | | 
| Human. | Ox. | Guinea-pig. | ’ 
| Organic constituents —. , 34:56 32°02 34°70 | 
| Inorganic constituents . : 65°44 TE 67 98 65°30 


NorMAL Unpriep Bons (Hoppe Sryner). 


Oy 1 ; . 11°90 
| Co Gas oh. eke 
Bone earth. ; a Ailes CG, ; ee 
| Fl ; : : Di 
L GH : . ; 04 
Rate a f : Sosa 
Proteid : eelIEA0 
Water : : . 50-00 
Organic =27°1i 
Inorganic = 71°85 
COMMENTARY. 


Besides the tables which give my own results I have appended Zalesky’s 
analyses of normal dried macerated bone and Hoppe Seyler’s analyses of 
normal undried bone.! 

Comparing the results of my analyses of the fossil seal bones with the 
normal figures, it will be seen that these fossil bones do not differ greatly 
from normal bone as regards the relative amounts of organic and inorganic 
substances. For the scapula the figures are 21°34 per cent. and 78°66 per 
cent.: for the astragalus 29-26 per cent. and 70°74 per cent.; for normal un- 
dried bone 27:15 per cent. and 71°85 per cent.; for normal dried macerated 
bone about 54 per cent. and 66 per cent. respectively. I have no analyses 
of normal seal bones with which I could compare my results, but it is not 
unlikely that they would resemble fairly closely the normal bones of other 
vertebrate animals. The seal bones resemble normal dried bone more 
closely than they do the normal undried bone, the seal bones containing 
only about 9 per cent. water, whilst normal undried bone contains 50 per 
cent. Their organic matter is well preserved. This was perhaps to be 
expected from the characteristics of the clay in which these bones were 
embedded. 

The humerus of a prehistoric man, which was found in sandy loam 
near North Berwick and therefore not very far from the spot where the 
seal bones were dug up, is not as well preserved as these; it contains 
an appreciable amount of organic matter, but much less than the seal 
bones. 

The proteids of bone resist disintegration more beatin: | than the 


1 Halliburton in Schiifer’s Text-book of Physiology, vol. “ape elu 


The Chemistry of Fossil Bone 133 


fat: normal undried bone contains 15°75 per cent. of fat and 11°40 per cent. 
of proteid, whilst the prehistoric humerus contains more than twice as 
much proteid as fat. This point is emphasised by the seal bones, which 
contain ten times more proteid than fat. Of course the fat is less closely 
bound up with the earthy material in the bone than is the proteid, most of 
the fat belonging tothe marrow. ‘The proteid in the seal bones and humerus 
was collagen. Mucoid was not present, but traces of a protein (nucleo- 
protein /) were observed. 

Turning now to the rib of Diprotodon, we note that there is no organic 
matter at all, although the bone belongs to the Pleistocene period. As 
already stated, the rib looked and felt like stone. ‘The climate of the 
Darling Downs, where the bone was found, is very hot in summer and cold 
in winter. These conditions hasten disintegration of organic matter. Fair 
quantities of silica, alumina, and manganese are present in the ash. These 
have evidently come from the surrounding soil. Fremy,' who examined 
a number of fossil bones of animals, found that the organic matter tends te 
be displaced by the substances (silicates, lime, ete.) in the surrounding soil. 
My results confirm this observation. 

I wish to express my indebtedness to Mr James Shelton, acting 
assistant Government analyst, Singapore, for advice regarding the gravi- 
metric methods. 

Part of this work was performed during my tenure of the Crichton 
Research Scholarship, University of Edinburgh. 


1 Ann. de Chimie, sér. 3, tome xliii. p. 47. 


OBSERVATIONS ON THE EXCITABLE CORTEX OF THE 
CHIMPANZEE, ORANG-UTAN, AND GORILLA. By A. S. F. 
LeyTon and C. S. SHERRINGTON. (With thirty figures in the text.) 


(Received for publication October 4, 1916.) 


CONTENTS. 


PAGE 
I. INTRODUCTION, AND Mrruops Empioyep .’ ; : : 135 
I]. EXPERIMENTS BY STIMULATION ; ‘ : ‘ 137 


1. Prefatory remarks on “localisation” of points in the motor cortex - 
functional instability of the motor points as evidenced by facilita- 
tion, reversal, and deviation of response ; , : : 137 
2. List of motor responses observed, and the topography of their cortical 
points ; “buried ” portion of the motor cortex ; remarks on the group- 
ing of the cortical motor points ; inferences as regards the functions 


of the motor cortex. c : : : : : 144 
Il]. EXPERIMENTS BY ABLATION . : 4 : : : : 180 
Protocols of experiments and degenerations of the pyramidal tracts 
observed : : 7 ; d ; : : 180 
Remarks on the ablation experiments. ; : ; 206 
IV. EXPERIMENTS ON GYRUS CENTRALIS POSTERIOR , 2 , 3 208 
V. StimunaTion oF SurFACE oF INSULA : : : : : 212 
VI. CorticaL THRESHOLD TO FARADISATION IN ARM AREA OF Car. MACAQUE, 
AND CHIMPANZEE COMPARED ; : ‘ ‘ : F 212 
VIL Inriuence or Locat Cotp aNp WARMTH APPLIED TO SCALP ON TEMPERA- 
TURE OF CORTEX (CHIMPANZEE) : : : : ; 214 
VIII. CrosurE oF CarotTip ARTERIES AND EXCITABILITY oF CORTEX 
(CHIMPANZEE) . : : / : : : ; 215 
IX. LocanisaTion or FIBRES OF THE PyRaMIDAL TRACT IN CRUSTA (ORANG) 
AND IN Pons (GoRILLA). ; : ; ; : ; 216 
X. SuMMARY OF ConcLUSIONS . : J : : : F 218 
REFERENCES . ¥ : : ; : : : : : 221 


lL. INTRODUCTION. 


THE investigation the results of which are here recorded arose from an 
observation, which chance opportunity afforded us, of examining by stimu- 
lation the cerebral cortex of a chimpanzee. That anthropoid species had 
not at that time come under experimental examination. On faradising the 
cortex we found, contrary to our expectation, that, although the gyrus 
centralis anterior yielded motor responses readily, we obtained none such 
from gyrus centralis posterior. A second similar opportunity arising, we 
repeated our experimental tests, and the results confirmed our former ones. 
Obtaining then a specimen of gorilla, an anthropoid also not previously 
experimented on, results were again met confirmatory of our first. It was 


136 Leyton and Sherrington 


therefore decided to carry out an inquiry into the motor cortex of the 
anthropoid apes, more especially from the “localisation” aspect. The 
following paper is based on the experimental examination of twenty-two 
chimpanzees, three gorillas, and three orang-utan. The methods employed 
have included both stimulation and ablation, but chiefly the former. 

At the time our observations were begun the only recorded experiment 
on the cerebral cortex of the anthropoid ape was one of stimulation of the 
cortex of an orang by Beevor and Horsley (1890) (2). The results they 
arrived at will be referred to later in the present paper; an excellent 
diagram summarising them is given by Schafer in his Text-book of 
Physiology (1900) (387). More recently, observations on localisation in the 
anthropoid have been (taking them in their successive order of date of 
publication) two preliminary Notes by ourselves (19); observations on the 
orang by Roaf and Sherrington (36) and by the Vogts (44): on the 
gibbon by Mott, Schuster, and Sherrington (32): on the chimpanzee by 
T. Graham Brown and Sherrington (5, 6): and on the chimpanzee by 
T. Graham Brown (3, 4). Individual reference is made to these sub- 
sequently in the text. 


MeETHODS EMPLOYED IN THE PRESENT RESEARCH. 


For stimulation of the cortex we have used faradisation, applied for the 
most part by the unipolar method (18, 19). For this a broad copper plate 
was strapped over a pad wetted with strong sodium chloride solution lying 
against the sole of the foot contralateral to the hemisphere under examina- 
tion. The pattern of electrode used was that figured in the Journal of 
Physiology, vol. xxviii. p. 16 (18). It has the advantage of being easily 
applied with a light and fairly constant pressure against the cortex surface 
without risk of pricking the cortex or its pia; also of being easily steril- 
ised by the flame, and of being readily bent to any appropriate curve when 
surfaces not otherwise easily reached have to be explored. The inductorium 
was of the usual physiological pattern, worked by a single Daniell cell. In 
many instances we used also the bipolar method, the electrode tips being 
2 mm. apart. The unipolar method is preferable, and gives minuter 
localisation, especially where, as in certain experiments, a cut surface is 
to be explored for fibres running at right angles to that surface. 

The animals were in all cases deeply anzsthetised with chloroform and 
ether mixture for the whole of the operation by which the cortex is exposed. 
During the actual exploration with faradism the anzesthesia was lightened, 
since in profound anesthesia the cortex becomes inexcitable. 

After the dura mater was opened it was always necessary to prick or 
tear some small holes in the arachnoid to let out the subarachnoid fluid. 
If that is not done, localisation in the neighbourhood of the sulci is almost 
or quite impracticable. 

A precaution found necessary for success in a prolonged examination of 


re . . ‘ . “1° - ° 
Che Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 137 


the cortex is prevention of a fall in temperature of the exposed cortical 
surface. The temperature of the room was therefore always kept high, 
usually fully 30° C.; and the cortex was kept as far as possible covered 
with cotton-wool swabs wrung out after being soaked with Locke's fluid 
at 38° C. 

For recording at the time of the experiment the localisation of the 
points stimulated and the responses obtained, the following plan was 
adopted. When the cortex had been exposed after the subarachnoid fluid 
had been evacuated, a thin glass plate warmed to rather above blood 
temperature was laid over the exposed cortex. The lines of the sulci were 
then traced on the glass, and also the lines of the larger arteries and veins. 
The drawing on the glass was then traced on millimetre-squared tracing 
paper. On this map, as the point-to-point examination of the cortex pro- 
ceeded, an observer then wrote numerals indicating each point stimulated 
at the point on the map corresponding with the point stimulated. Another 
observer listed the responses obtained, entering each response on his list 
opposite a numeral corresponding with that locating on the map the posi- 
tion of the point which evoked it when stimulated. 

Ablations were performed with the low-tempered knife recommended 
by Mott; the blade of this knife can be bent to any curve desired at the 
time. A number of the ablation experiments were performed under aseptic 
precautions of the usual kind and the animals allowed to recover. Their 
details are given below under the separate experiments. 

The animal used was for the most part the chimpanzee, Anthropo- 
pithecus troglodytes. Among the specimens was one of the bald variety, 
A. trogl. calvus, the variety to which belonged the well-known “Sally,” 
observations upon which were made by G. J. Romanes. Two others were 
of the variety known as Kola-kaamba by the dealers. The convolutional 
pattern of the Rolandic region of the hemisphere of our specimen of Calvus 
is given in fig. 6, B; v. infra. Besides the chimpanzees, three orang-utan 
(Simia satyrus) and three gorillas (Gorilla savagei) were used. 


Il. EXPERIMENTS BY STIMULATION. 


1. Prefatory Remarks on the Motor Responses obtained 
and their Localisation. 


Regarding the subjoined list (p. 148) of movements obtained in response 
to localised faradisation of the cortex, they are recorded in all cases in the 
notation made at the time by an observer entrusted solely with the 
observation of them, the management of the stimulation and the recording 
of the point stimulated upon the previously prepared map being in other 
hands for that time. In some cases there appear in the list notations 
which are not detailed, e.g. “mouth”: such instances mean that the 
observer, though seeing that a movement of the mouth had occurred did 
not feel able to say what that movement had been exactly, and that on 


138 Leyton and Sherrington 


repetition of the stimulus he did not feel sure that the movement then 
obtained was the same as that seen previously. In some instances words of 
description were entered down which were equivocal, thus “ankle flexion,” 
leaving it uncertain whether the movement was dorsal flexion or plantar 
flexion. But these imperfections where occurring have not been allowed 
to exclude the observation from the list. 

The main object in view being to “ localise” the motor function of each 
cortical point yielding motor responses, the stimuli applied were systematic-* 
ally kept of weak strength and not far above threshold value, and each 
stimulus was applied usually quite briefly, e.g. 1’—2” or little more. The 
sequences of movement are therefore short, our intention being to determine 
chiefly the primary movement yielded by the cortical point. 

We had supposed at commencement of our experiments that the 
identification of exactly corresponding points in the two hemispheres of an 
individual and in the hemispheres of different individuals could be much 
more nearly possible than our experience has left us with the impression 
that in fact it is. The dissimilarity of the convolutional pattern of the 
hemispheres even in individuals of the same species (Troglodytes niger), 
and the seemingly variable relation of analogous functional points to sulei 
of corresponding name, makes it practically impossible to decide with 
sufficient exactitude what point on the hemisphere of one individual is 
identical with a given point upon another hemisphere. But in spite of 
this inability to determine what point on one hemisphere is anatomically 
identical with some particular point on another hemisphere, our series of 
experiments as they proceeded, each resulting in a detailed localisation 
map, showed us clearly that in very many cases, probably in most, the 
corresponding anatomical points in different individual hemispheres did 
not, as examined by faradisation in the course of experiment, yield motor 
responses so nearly similar as to be noted as the sanie movement in our 
movement list. Of this many illustrations can be found if the maps with 
number references furnished in this paper are turned to. The movements 
so obtained were related movements, often or indeed usually closely related 
movements, but not identical, not rarely movements of opposite sense, 
although of the same part. And many instances may be found in the 
maps where one and the same movement, as noted by the observer, was 
obtained in one hemisphere from some point which was clearly not that 
at which it was obtained in another hemisphere, either the opposite hemi- 
sphere of the same individual or the hemisphere of the corresponding side 
in another individual. Our experience is thus clearly in harmony with 
that of Shepherd I. Franz (17) on the macaque. Summarising the 
observations described in section ii. of his paper, he writes: “The data 
show in different animals,” i.e. different individuals of the same species, 
Macacus rhesus, “and in different hemispheres a variety of distribution 
of the areas concerned with the movements of the individual segments of 
the leg and arm.” 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 139 


And it was clear, in our experience, that the motor cortex of an 
individual hemisphere and of both hemispheres in one individual does not, 
as its surface is gone over point for point in a systematic localisation 
experiment, yield the whole series of movements that can be yielded by 
similar examination of a series, even a small series, of hemispheres. 
Movements will appear in one hemisphere which do not appear in another, 
or, putting it in another way, will appear in one experiment which do 
not appear in another experiment. We think that this is probably 
largely owing to “facilitation.” When the motor cortex in any individual 
hemisphere is systematically explored point to point by the electrode, 
particular motor responses when once evolved tend to reappear from 
adjacent cortical points. These cortical points form groups, each group 
occupying a small cortical area whence the same motor response is elicited. 
Such an area is probably partly the result of the facilitation exercised in 
regard to the response characteristic of it by the influence of one point 
upon another in it. This facilitation of one response would act in the 
direction of restricting the appearance of other responses which neverthe- 
less might be latent in the cortex: it would tend to deviate the response 
(v. infra). 


Functional Instability of Cortical Motor Points. 


This raises the question of the functional instability of a motor cortical 
point (5). In addition to the influence of depth of narcosis, freedom of 
blood supply, local temperature, and such effects of experimental exposure 
of the cortex as “drying” or inspissation of applied Locke’s solution, the 
motor responses of a cortical point may be easily and greatly modified by 
precurrent, especially closely precurrent, stimulation either of itself or 
of neighbouring, especially closely adjacent, cortical points. The motor 
response from a given point, though it may, as the maps of cortical localisa- 
tion usually depict, remain approximately the same throughout a lengthy 
experiment, even from hour to hour, when similar stimuli are repeated at 
intervals not too brief, may yet vary considerably in result of precurrent 
stimulations not too distant in time and place. Experiments in which a 
large field of cortex is examined systematically point for point by electrical 
stimulation to determine the functional localisation are likely to display 
the influence of previous stimulation of one point upon another. 

Three phenomena of this kind, presumably all closely akin, make them- 
selves evident in an examination of the motor cortex, namely, facilita- 
tion of response, reversal of response, and deviation of response. 
Of these the first, noted by various observers (e.g. 15), and particularly 
fully studied recently by T. Graham Brown (3, 4) in the chimpanzee as 
well as in macacus and other monkeys, is characterised by a change of the 
cortical point’s response in the direction of increase, with or without other 
modification. It may be induced by stimulation of the point itself or by 
stimulation of other points. Reversal of response (5) is a change super- 


140 Leyton and Sherrington 


vening which may culminate in complete reversal of the sense of the 
movement of the response, e.g. extension of a joint may become flexion of 
that joint. Deviation of response is a change which alters the character 
of the response, so that instead of the original movement appearing, some 
other movement, e.g. of another joint or part, appears in place of the 
original. All these changes are temporary. They may be taken as 
expressions of what has been termed the functional instability of a cortical 
motor point (5). 

1. Facilitation of Response.—Facilitation of response has been a 
usual accompaniment of our observations on the anthropoid motor cortex. 
Comparing our experience of it there with our experience of it im macaque 
and ealothrix, facilitation seems to be somewhat more extensive in the 
anthropoid than in the lower forms of monkey. Thus, as was remarked 
in our preliminary communication, it affects the delimitation of the whole 
of the anterior border of the motor field. That border is not of sharp 
and abrupt edge, but seems to fade off forward rather gradually. Facili- 
tation makes it extend farther forward than it does without facilita- 
tion. Thus if the anterior border is delimited by stimulating series of 
cortical points in succession from behind forward, the anterior limit of 
the field is found to le farther anterior than if determined by stimulating 
a series of points starting well in front of the limit and followed from 
before backward. In a similar way the boundary of the area for any 
particular movement may by facilitation be extended beyond its average 
limit; in this latter case, deviation of response comes in as well. 

2. Reversal of Response.—The mutual influence exerted by points 
moving the same joint but in opposite directions was dealt with in the 
paper by T. Graham Brown and one of us (5). 

3. Deviation of Response.—A cortical point can also influence the 
motor response of another whose response is neither diametrically opposed 
to nor identical with or very closely similar to its own. ‘Thus: chimpanzee 
(19), left hemisphere (fig. 14), leg area. Point 519 gave regularly as 
response plantar flexion of ankle, followed by flexion of all toes except 
hallux, followed further by adduction of hallux. That was its response 
when first stimulated in the experiment. It was the fifth point stimu- 
lated in the experiment, and was then stimulated next after a point 41 
in face area, which yielded movement 41 (see list); and point 41 had 
been stimulated next after one in arm area, which yielded movement 282. 
From time to time in the course of the experiment point 319 was returned 
to from distant points, and gave as response regularly movement 319 and 
no other. 

Point 342, similarly stimulated, was giving with like regularity move- 
ment 342, flexion of hip followed by adduction of hip. When point 319 
was stimulated immediately after point 342 had been stimulated, time being 
allowed, however, for the movement evoked from 342 to subside completely, 
the movement given by 319 was no longer movement 319. It then gave 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 141 


instead flexion of hip followed by dorsal flexion of ankle and flexion of 
knee, followed further by flexion of all toes except hallux, movement 363. 

Again, in the same animal and same hemisphere, point 306 gave regu- 
larly, when stimulated after points relatively indifferent to it, the response : 
flexion of little and 4th toes rapidly followed by flexion of 3rd and 2nd, 
and then followed by plantar flexion of angle, and later adduction of 
hallux. When stimulated next and quickly after point 306, but with 
allowance of time for the movement from 306 to subside completely, 
point 319 evoked no longer movement 319, but the following: flexion of 
toes without hallux, followed by plantar flexion of ankle, followed by 
adduction of hallux, movement 298. 

Again, point 268 gave as its regular response flexion of hallux, followed 
by flexion of toes, followed by plantar flexion of ankle. Point 319, when 
stimulated next and soon after 268, evoked as response, simultaneous 
flexion of toes and hallux, followed by plantar flexion of ankle. 

Again, point 331, which evoked flexion of knee when stimulated in 
quick succession to point 342, evoked when stimulated in quick succession 
to point 263 extension of hallux, the regular response from point 263 itself 
being extension of hallux, followed by extension of the remaining toes, 
followed further by dorsal flexion of ankle, and finally by flexion of hip. 

Again, point 232 in the same experiment, in arm area, yielding 
ordinarily flexion of elbow, yielded when stimulated quickly next after 
point 127, which was yielding flexion of thumb, flexion of thumb followed 
by flexion of elbow, 140. 

Again, in another animal, the following “deviation” occurred. The 
examination of the motor area had in this experiment been begun at the 
top limit of the arm area in shoulder region, and proceeded systematically 
from point to point in the downward direction. Followed in this manner, 
elbow flexion soon became the leading (primary) movement, and continued 
so very nearly or even quite down to the inferior genu of sulc. centralis. 
Beyond a certain point, which was minutely and precisely marked on the 
map made, elbow flexion disappeared abruptly, and facial movements 
appeared in the form of closure of opposite eyelids. The lower margin of 
arm area having thus evidently been reached, we turned to the delimitation 
of the face area. The examination of this area we started at the lower 
(Sylvian) end of sule. centralis, and thence proceeded point by point up- 
ward along the precentral gyrus not far in front of sule. centralis. In due 
course the point yielding closure of opposite eye was again reached, and it 
was found that then on proceeding farther upward to the point that had 
previously yielded elbow flexion as its primary movement, that point now 
yielded adduction of thumb as its primary movement, and a little farther 
upward movement of index, chiefly extension, was added to that of thumb: 
and movements of thumb and index continued to be the primary move- 
ments right up through the region which previously had given elbow 
flexion as primary response, and thumb and index movements as primary 


142 Leyton and Sherrington 


responses trespassed actually into the area that had previously yielded 
shoulder movements as the primary response. Here the “deviation of 
response” was seen to affect a whole series of points, influencing in its 
special direction a not inconsiderable fraction of the whole arm area. 

Again, in an experiment on a gorilla, a point 172 which had been 
yielding regularly flexion of all fingers without thumb, on being stimulated 
next after a point 142, which yielded extension of index finger alone, 
yielded extension of index alone without movement of the other fingers 
(fig. 12, A, points 172, 142). Later, when stimulated after an interval of 
some two minutes, it yielded flexion of fingers as at first. 

Again, in the same experiment, a point 176, which yielded regularly 
extension of fingers without thumb, on being stimulated in next succession 
to point 172 yielded flexion of fingers instead of extension of them. It 
was, however, not found possible by stimulating point 172 in next 
succession to point 176 to obtain movement 176 from point 172. Further, 
on stimulating the two points, by separate electrodes, concurrently it was 
found that a stimulation of 172, weak as judged by the induction scale 
and also by its nearness to threshold value of excitation, caused flexion 
of fingers in spite of concurrent stronger stimulation of 176. Also an 
extension of the fingers already brought about by stimulating 176 alone 
was broken down and converted into flexion by weak stimulation then 
applied to 172, although the stimulation of 176 was continued unremitted. 
Indeed the extension of fingers produced by stimulation of 176 seemed 
more readily broken down and changed to flexion by stimulation of 172 
when the stimulation of 176 had been in progress for some little time than 
when stimulation of 172 was introduced earlier. When, conversely, flexion 
of fingers was in progress under stimulation of 172, the application of 
strongish stimulation to 176 broke down the flexion and changed it to 
extension; but for this the stimulation of 176 had to be strongish. Also 
when the stimulation of 172 and 176 was commenced concurrently, but 
the stimulation of 176 was strong and that of 172 very weak, the result 
obtained-was extension not flexion, that is, 176 overpowered 172. 

Again, we have in several hemispheres observed that a cortex point 
which ordinarily evoked as its primary response flexion of the elbow 
would evoke, when stimulated next and soon after a distant point giving 
adduction of thumb, flexion of elbow and adduction of thumb. 

Again, on the opposite hemisphere of the same gorilla referred to above 
a point 218, which yielded regularly flexion of wrist followed by flexion of 
elbow, yielded on being stimulated after stimulation of a point 251 yield- 
ing rotation of shoulder, rotation of shoulder and not flexion of wrist or 
elbow. It yielded these latter, however, as secondary movements if the 
stimulation were prolonged, and it yielded them again as its primary re- 
sponse after a time interval had been allowed (fig. 12, B, points 218, 251). 

The distances across which “deviation of response” may be exerted by 
one point on another vary. Though usually the space intervening between 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 148 
the points is short, e.g. less than 5 mm., it is sometimes double or treble 
that. It is not equally developed in all directions, thus it tends to occur 
more readily between two points situate in one and the same functional 
area, e.g. leg area, than between two points situate in different functional 
area, e.g. one in leg area and one in arm area. It seems to occur more 
readily between points which under stimulation give rise, in the “march” 
(Hughlings Jackson) elicitable from them, to similar motor responses. 
Thus in the march elicitable from “abdominal wall” points movement of anus 
is prone to occur, and, vice versa, in the march elicitable from “ anus” points 
there is a proneness for abdominal wall response to appear: and similarly 
between anus points and abdominal wall points, though their foci are situate 
quite far one from the other, we have seen deviation of response exerted. 

Again, in an experiment a portion of the leg area was ablated, leaving 
below the ablated portion a small transverse slip of cortex which the faradic 
stimulations prior to the ablation allocated to hip area but abutting upon 
the abdominal wall area. The whole of this strip on being faradised twenty 
minutes after completion of the ablation yielded no trace of limb movement, 
but evoked instead vigorous contraction of the abdominal wall. Prior to 
the ablation it had yielded as movement chiefly flexion of hip; after the 
ablation it yielded contractions of the contralateral abdominal wall without 
any movement of hip. 

Again, in a chimpanzee, at the region of the gyr. cent. anterior, opposite 
the brachio-facial genu of sule. centralis, the following was noted. The lower 
limit of hand area was determined, care being taken to avoid as far as 
possible deviation of response by near precurrent stimulation of adjacent 
points. Similarly the upper limit of angle of mouth area was delimited. 
Then the lower limit of hand area was obtained by stimulation in serial 
succession of a number of points descending in order from upper part of 
arm area downward. The lower border of hand area as thus examined 
trespassed into face area according to the upward limit of the latter as 
demarcated previously. The responses of hand given by the hand area 
points thus trespassing were always similar to the last hand responses 
obtained from the portion of the hand area above them; and they were 
accompanied by “angle of mouth” movement, either simultaneous with 
them or almost so. Conversely, on determining the upper limit of angle 
of mouth area by following that area upward along a series of points 
stimulated in it in turn, the upper limit trespassed over into hand area. 
The responses of mouth movement from these trespassing points always 
resembled the mouth responses last obtained from points lower down in 
mouth area, and were accompanied by movements of hand. Similarly, at 
lower edge of closure of eyelids area that area could by serial stimulation 
of it be made to encroach on “angle of mouth” area, which lay lower down 
and rather posterior to it; and the upper and posterior edge of closure of 
eyelids area could be made to encroach over into hand area, which lay 


above and rather behind it. 
VOL. XI., NO. 2.—1917. 10 


144. Leyton and Sherrington 


The above instances are cited as typical and somewhat outstanding 
examples of what in a smaller and less pronounced manner was frequently 
met by us. As to how far such deviations, as also the reversals and facili- 
tations, are traceable to shuntings of route in the cortical structure itself, 
or how far they are referable to shuntings in sub-cortical paths and 
centres, that is a question towards whose solution our observations contri- 
bute little or nothing. The diagram furnished by Franz (17) (fig. 16, 
p. 148 in his paper) indicates the manifold possibilities in that respect, 
and Graham Brown (3, 4) has published direct observations in regard to 
“facilitation” which throw light on the problem as concerns that pheno- 
menon. ‘The main point that we wish here to emphasise in preface to the 
subjoined list of motor responses and maps of cortical points belonging to 
them as observed in our experiments is, that in looking through such data 
we would wish the reader to bear in mind that the fixity of such localisa- 
tions is as regards minutiw to some extent probably a temporary one, i.e. 
obtained at the time of observation, but in our opinion might not be pre- 
cisely the same were examination possible at a number of different times 
and in a number of different experiments. As regards minutiz of localisa- 
tion in the motor cortex, our experience agrees with that of those (36, 5, 4, 
17, 3) who tind, as Shepherd Franz (17) expresses it, that the motor 
cortex is a labile organ. 


2. List of Motor Responses and the Topography of their 
Cortical Points. ; 

The maps (figs. 1-6, 8-9, 12-16, 19, 24, 27) which illustrate the “ List 
of Motor Responses” are all approximately life-size. Fig. 7 is on a some- 
what larger scale. The maps, except figs. 7 and 13, were prepared by 
placing a thin plate of glass upon the surface of the hemisphere and trac- 
ing the sulei upon it. The plate was first applied to the brain in situ over 
the exposed part of the hemisphere, and, where this did not afford sufficient 
of the surrounding part of the hemisphere, the plate was reapplied after 
removal of the brain, and the parts surrounding the originally exposed 
portion were added tothe map by tracing the sulci on the glass. To follow 
the sulci over the convexity of the hemisphere the glass was tilted succes- 
sively in the various directions required, and those of the sulci lying in 
contact with and slightly flattened by the plate were traced. An approxi- 
inately true plane projection of the convex surface of the hemisphere 
was thus obtained. On these maps the cortical points localised with their 
motor responses listed were copied in from the maps similarly obtained by 
the glass plate and filled in as the experiment proceeded. In the maps as. 
reproduced in illustration of this paper it has been impossible to include 
all of the points actually observed and recorded in the experiment, because 
the number of them written into the actual experiment-map was often 
large, and the numerals had therefore to be written so small as to render 
them illegible unless the size of the map were considerably enlarged im 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 145 
reproduction. In the maps as given here the size of the numerals has been 
enlarged, but the original size of the map as a whole retained. This has 
necessitated the omission of some of the numerals written in the originals ; 
the omitted numbers are for the most part those indicating responses 
similar or nearly similar to others at neighbouring points and included in 
the reproductions. In some of the maps the figure 0 is written at certain 
points to indicate that from those points no motor response was obtained. 
The numerals in the maps and plans refer to the numerals prefixed to 
the motor responses furnished in the List of Responses. The place of each 


33) 33! 
~ 331 33! 33/7 33) 53! 


Fie. 1.—A, chimpanzee 3, left hemisphere. b, chimpanzee 4, left hemisphere. 
The numerals refer to the ‘* List of Responses,” p, 148. 


numeral inserted in the map indicates the position of the spot whence, in 
the particular hemisphere from which the map was made, that response as 
listed was evoked by faradisation. The topographical item thus obtained 
has to be accepted as a datum for “localisation,” subject to the caveat 
entered by the prefatory remarks on the functional instability of cortical 
motor points. The maps show that the same numeral or letter, i.e. the 
same movement, does not always appear at the corresponding point of 
different hemispheres, either right or left. The letters C, O, G prefixed to 
the numerals in the list indicate that that particular response appears 
figured in one or more of the maps reproduced, chimpanzee, orang, and 
gorilla being signitied by the corresponding initial. The words “contra- 
lateral” and “opposite” mean always contralateral, or on the opposite 
half of the body to the hemisphere stimulated. 


Fic. 2.—Chimpanzee 6, A, left hemisphere ; the stippled area enclosed by the dotted lines together 
with the parts of the adjacent sulci exhibit the portion of cortex ablated (see ablation ex- 
periment 3, p. 194). The numerals inserted in the ablated area indicate the responses 
obtained before ablation, also numerals 262 and 281. The bracketed numerals (343) indicate 


the responses obtained from the same points at the final experiment. 1, right hemisphere 
of same animal reversed for comparison with left hemisphere. 


18: 
431° 16 


Fic. 3.—Left hemispheres of t 


wo very young chimpanzees 7 and 15, the figure B from the 
younger of the two, a baby. 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 147 


Fig. 5.—Chimpanzee 8. A, arm area of left hemisphere; the broken lines indicate the limits of 
cortical ablation at successive operations, January 3, March 3, and April 8. ‘The numerals 
mark the responses observed at the time of the operations. B, right hemisphere of same 
animal ; the broken lines indicate the limits of cortical ablations at successive operations of 
April 2and 3. The numerals mark responses obtained either at the times of operation or at 
the final examination. 0 denotes that no response was at any time obtained from the place so 
marked. In the upper part not all the responses are marked in. C, left hemisphere showing 
responses obtained at the final examination and the shrinking and deformation of the ablated 
area (see p. 186). 


Numeral or 
letter indicat- 


COTO Ot wm 69 FD 


9 


52 


ing response, 


148 


Appears in 
maps under 
anthropoid 
species, 
C, O, or G. 


G 


eolololo) 


c 


Cc 


OOO. =). 
(Oy Bod 


: OO 


OaAaaa: 


Leyton 


and Sherrington 


List oF Moror RESPONSES OBSERVED. 


lst movement. 


2nd movement. 


srd movement. 


Contralateral angle of mouth retracted 


” ” 3 ” 
’ ”? ” ” 
” ’ ” 
” 2° ” ” 
” 7) ” ” 
” ” ” ? 
” ” ’ ” 
” ” ” ” 


Lips parted chietly on contralateral 
side 
Lips closed a 


” ” 


Lips “withdrawn and compressed, 
chiefly contralaterally 


” 29 99 9 
39 32> ” ” 
Lips inverted 
Upper lip moved 
+] 2 
3 Cverted 
lifted and its edge inverted 
a retracted 
everted 


7 
Lips and tongue moved together 
Contralateral angle of mouth retracted 
Upper lip and nose moved downward 
Both lips retracted but the lower 
much the more 
Contralateral angle of mouth tightly 
compressed 
Lower lip raised 
» raised and retracted 


a pouted 

‘ee elisted 

»  Yretracted 

>  everted and protruded 
” ” ” 


tel 2) 
pouted, more on contra- 
lateral side 
Sucking action of cheeks with lip 
protrusion 
Lips and jaw opened 
Upper lip lifted and everted 
», protruded with slight pro- 
trusion of lower 
Mouth moved 
Lips and jaw closed 
Lips opened without deviation 
Lips and jaw closed 
Mouth closed suddenly with a snap 
Upper lip retracted to contralateral 
side 


» ” ” ” 


Both lips drawn far round to contra- 
lateral side 

Both lips and nose drawn to contra- 
lateral side 


” 9 


” : ” ” 


39 33 ” 33 
Contralateral angle of mouth retracted 


” oh) ” i) 


%” 29 %9 ” 

and nostril twisted be 

Both angles of mouth depressed, the 
contralateral the more 


| Jaw opened without deviation 


Jaw opened 


Contralateral nostril wrinkled 
pinna moved 
s, retracted 


” 
Lips opened 
Tongue moved toward ipsilateral side 
Front of tongue recurved 
Tongue thrust forward 


2 ” ” - 
Neck turned to contralateral side 


Contralateral angele of mouth retracted 


Tongue retracted 
Nose wrinkled 
Jaw closed 

Nose wrinkled 


Lower lip retracted 
Nose moved 


Arm moved 


Tongue turned to contralateral side 


Movement of the skin of the chin 
Upper lip lifted 

: retracted 
Sucking movement of both lips 
Tongue curled upward 
Twisting of contralateral nostril 


Tongue moved 


Elbow tlexed 


lloor of mouth depressed 
Jaw opened 


” 


Tongue protruded 


92 
Jaw closed with a snap 
Fingers flexed 
Contralateral eye closed 
5 nostril moved 


. eye closed 


”) on 


Tongue 


Palate raised 
Eyelids closed 


Neck turned away 


Tongue moved 


Tongue turned to 
contralateral side 


” 


‘Tongue turned to 
ipsilateral side 
Tongue curled to 
roof of mouth 


Contralat. pinna 

retracted & raised 

Contralateral eye 
closed 

Neck turned away 


9 


4th movement. 


Pinna moved 


Jaw opened wider 


Tongue to con- 
tralateral side 


Tongue protruded 
Lips closed tightly 


§ & & E#@RSSE 


Be 


Numeral or 
letter indicat- 
ing response. 


The Excitable Cortex of the C 


Appears in 
maps under 
anthropoid Ist movement. 
species, 
C, O, or G. 
| 
c;o G Tongue retracted and curled upward 
So} 0}... »» moved 
CO. G_ Sucking action of lips 
CO} G | Tongue retracted 
ro .. oy to contralateral side} 
a Se | “a fairly straight 
C . |G Tongue heaped up and twisted, bring- 
i ing dorsum to contralateral cheek 
© | .. |... Tongue retracted to contralateral side | 
’ 
Gi...) -G ? "2 + 
c|..|G - * c a 
c}o|.. es : 
ae . ” 
C|0/|G x 5 ms 
cC|0/|G . heaped up at back of mouth 
c 0 | 6 a 4 
Cc, 0;|G protruded 
Rat és posterior part de- 
/ ' pressed 
0 Be fe = toward ipsilateral 
} side 
EO ae . - re a 
an | OG ia = and twisted on et 
; long axis 
Cr. G + tipturnedtocontralateralside 
Cc Oo}... | ., protruded to ipsilateral side 
c Go . tip protruded 
Cc a | a = 
c | oO ' zs ‘ and makes a 
/ licking movement 
c;o | AC , tip protruded 
oh eee :, tipturnedtocontralateralside) 
eS ee protruded 
C | O | G | Hollowing of tongue tip 
C | O | G | Tongue twisted on its long axis, and 
dorsum pushed into ipsilateral cheek 
a hes | .. | Tongue, undeviated protrusion 
Cc = ee re twisted and curled upward | 
] to roof of palate 
CaO... aa curled upward  withont 
lateral deviation 
C |} O| .. | Hollowing of tongue tip 
C |... .. | Tongue retracted 
CW Ory)... ,. tip curled upward to touch 
| roof of mouth 
ON a Ta .. protruded to ipsilateral side 
Or ..| G ., tip protruded with narrowing 
of tongue 
¢} 0 ; tip protruded and curled up- 
ward 
C | .. | G | Contralateral side of soft palate moved 
€ | 0 | .. | Sucking action of lips and tongue 
Cc} .. | Uvula drawn to contralateral side 
C | O.| .. | Lower jaw drawn to contralateral side 
Te Met, G Pp depressed with lateral de- 
viation 
Coe | he “ depressed without lateral 
deviation 
Cc ao dis 7 2” 79 ” 
Cog eee | a 2 Ss - 
Cc los As depressed without lateral 
deviation and wholefloor 
of mouth depressed 
Cc Jaw opened and tongue made a click 
} against roof of mouth 
Cc © po ” 2 ” 
¢ 6G and tongue curled up and 
| sucking noise made 
Cc _ G | Sucking action of the cheeks 
eh | .. | Jaw lifted 


himpanzee, Orang-Utan, and Gorilla 149 


2nd movement Srd movement. 4th movement 


| 
Upper lip moved | Nose moved 
Jaw opened 
Lips move 
Jaw closed 


Lips retracted 


Masticatory 
movements 


Jaw opened Lower lip to con- 
tralateral side 
| Jaw opened and drawn to contra- 
lateral side 
_ Lower lip drawn to contralateral side 
| Upper lip raised 
iC ontralater: al angle of mouthretracted 
Bilateral adduction of faucial pillars 


Bilateral adduction of faucial pillars Lifting of hyoid 


| Tongue retracted Tongue tip curled) 

| down 

| of and heaped up Faucesand glottis Closure of jaw 
closed 


| Tongue retracted and tip curled down | Fauces and pz ulate, Jaw closed 


moved 

| Tipe parted i 

Base of tongue depressed Faucial — pillars 
moved ! 


_ Lips retracted 


Contyralat. angle of; 
| | mouth retracted 
Lips opened and pouted | Jaws open | 
‘Tongue retracted ;, closed | Lips pursed 
| Opening of lips Licking movement) 
| 


Tongue retracted 


Contralateral angle of mouth retracted Palate raised 
Tongue turned to contralateral side Jaw opened 
| 


Tongue tip turned to opposite side | 
Contralateral nostril flattened and | 
| depressed 


Tongue retracted to contralateral side 

Tongue tip to roof of mouth on 
opposite side 

me turned to ipsilateral side | 


Tongue protruded straight } 
ee - | Tongue retracted 
to contralat. side | 


3° ” 


Larynx moved as a whole 


4th movement. 


150 Leyton and Sherrington 

os 2 Appears in | 
SSS) maps under | 

5-5 anthropoid | Ist movement. 2nd movement. | 3rd movement. 
aes species, 
ase | €, 0, or G. 

112 as Cheeks (contralateral) drawn up Contralateral nostril raised 

118 (Ol Seen hae an op flattened 

114 © | O | G | Chewing movement 

115 Calne . | Tongue curled upward 

118 c ) O} ., | Thumb extended 

119 G0) | ze Ls Index extended 

120 CO | -¢E ,  adducted 

121 Co) ull nA - and fingers tlexed ‘ 

122 ¢ | G > extended Index extended Other fingers ex- 

tended 

123 Cl}... .. », adducted Wrist flexed Fingers extended 
124 Cale ee i 7% a » flexed 
125 C = Fingers and wrist moved together 

126 sll aoc oa prs i) etlexed Elbow flexed 
127 Cc}; 0 sutexedl 

128 C hike oe : Fingers tlexed 

129 c Gi . Index flexed Other fingers 

| \ flexed 

130 OHO! | Gt 7 .. and adducted +y 

131 Cole || 8G - A re Bs Kingers tlexed Wrist flexed 
132 (Clee | Gea =e ie oe if Wrist extended 

133 C10) Gi 5, extended Thumb adducted ce 

134 c lecend ee im Fingers extended Wrist extended 
135 ae 1G 3 Wrist abducted 

TSGGE Alea ee nGeuly ie Thumb adducted Index flexed 
137 Cc | eae ts es = BN | Fingers flexed 
138 (0 | 3s % Wrist flexed 

139 (0; | H », abducted ,, extended 

140) Cc om » flexed Elbow flexed 

141 @lltse |) ge | - - Index extended 

142 | C |} O | G | Index extended 

143 | Cal Na hece Ao terminal phalanx only 

144 | C] } 5 ie Wrist extended 

1450 (ec) | : as Index tlexed Thumb flexed 
146) |) (C se - 5 ae Wrist flexed 
1470 ala G “i 9 Wrist supinated 

148 -C] G oF as Middle finger extended 

149 «| Ae 3 ~ Thumb adducted 

150 | C G » flexed ms) 

151 C G a ms Thumb adducted Thumb tlexed 
152 C | 50 56 Wrist flexed 

158 a (ive #3 | Other tingers flexed 

154 Cc G » and middle finger tlexed together, 

155 Cc I 3 rf , extended ,, | Wrist flexed | 

156 (Os » extended Thumb abducted | 

157 (oh | and middle fingerflexed together, Wrist flexed 

158 ee || ,, thumb flexed 

159 ( extended Extension and spreading of other | 

| tinger's 

LOOM SCs iG 33 Thumb extended 

161 | C| O.| G | Middle finger extended | 

162 | C 5 »  andring-tingerextended 

163 | € | Little finger flexed | 

164 | C 4s os vs Other fingers extended 

165 | C “ on 50 An flexed 

166 | C Pa » extended 

167 =| C IS #5 e | Wrist adducted 

LOSI | iretei| i E ,, and ring-finger extended 

169 | C} G eS ee x flexed 

170 Cal Res es ae Ka 4 extended | 

171 =| C | O| G | Alitingersextended withoutthethumb, Middle and index extended 

M2 CO : 3 flexed without thumb | 

1738 C ‘ 3 a a Wrist flexed 

17 C j os a i » extended and pronated Elbow flexed 
175 CO ene 4| - * oe ae .,  pronated 

76y | CuO) a extended without thumb 

177 30 bs 0 3 5 a | Wrist abducted 

178 C | G 5 “; | ., extended 

179 Cc | 5 3 BS se | Fingers flexed Wrist flexed 
180 C | 3 55 * 50 Thumb adducted Fingers flexed 
181 ¢€ lets i 1 ‘5 Elbow flexed | 

182 C G | All fingers and thumb flexed Wrist flexed | 

1838 C -, flexed without thumb | Wrist pronated Thumb flexed 
184 Cc re 55 5 a Thumb flexed Thumb adducted 
185 Cc > 1) a 5 Fingers extended 

186 Cc ; or , Wrist extended 

187 C , o 55 on Wrist flexed Wrist supinated 


Elbow flexed 


Numeral or 
letter indicat- 


255 


ing response. 


The Excitable Cortex of the Chimpanzee, Orang-Utan, 


Appears in 
maps under | 
anthropoid 
species, 
C, O, or G. 
cio 
hss | 
oy. 
c / 
C . 
C G 
Oo G 
C a 
Co — 
Cc “ 
GC G 
c G 
C10; G 
C10 )'G 
ee G 
Cc G 
op G 
c iG 
c G 
‘2 bra ee 
canal .. 
Sealer, iss 
C | 5 
Gt. 1G 
cio : 
Cc : 
Cc ; 
c G 
C G 
Cc 
Cc 
oe a 
c/o 
Cc 
cel ae 
ie’ yy: 
Car .. | G 
oS 
Sales. A 
a) ° 
Os. 
nO.) G 
Cc} Q;G 
Cc; o0o|G 
Ces |. 
alc.) G 
CaO]. 
clolé 
CeO! |e: 
cio 
C Vere G 
Cars. iG 
OM een FE 
Gel, | G 
Cc} ..| G 
(CA aaa $i 
ot seid IE 
c/0o0;|G 
aes. 
Pr | G 
Cc | G 
Cc bs 
c/o} 
C | 
C 


| 
| 
| 


Chree tingers of ulnar side of hand 


| Opening of whole hand 


Ist movement. 


All fingers and thumb tlexed 


" 


a + * 
ve flexed without thumb 
and thumb flexed 


a ” 


» - spread widely | 
Three fingers of ulnar side of hand | 
extended | 
Three fingers of ulnar side of hand | 
flexed 


” ” 


” ” on ” 


; 
extended 
Ulnar side of hand extended and. 

adducted 
Closing of whole hand 


” ” ” 
Closing =: . 

33 - ». beginning with 
ulnar side 
The three ulnar-side fingers flexed 
and index extended } 

Wrist flexed 
+»  pronated 
»» flexed 


ce) 


adducted, ie, to ulnar side 


Lal ” ” 
. extended 

” ” 

oa - 

bal 99 

», Hexed \ 
BI ” 

? be) 

” ” 

” ” 

9 ” 

»  supinated 

” 9 

,.  pronated 

5, supinated 

” ” 


”? . . . 
,, adducted, 7.2, to radial side 
Elbow flexed 


” ” 
” ” 
39 
», extended 
rt ” 


Shoulder raised 
retracted 


” ” 


abducted 


adducted 
retracted and abducted 


” 2? ” 
raised and protracted 


rotated inward 

ef + outward 

and muscles of back moved 
(contralateral) 

and muscles of front of chest 
moved (contralateral) 

and muscles of abdominal 
wall moved (contralateral) 

lifted 


2nd movement. 


| Wrist extended 
| Wrist pronated 


Thumb adducted 


+ 99 
Interosseous extension of digits 
Wrist supinated 


Index tlexed 
Wrist flexed 
Wrist adducted 
Index extended 


Wrist supinated 


Wrist flexed 
Wrist extended and supinated 


Elbow extended 


Wrist pronated 


| Wrist pronated 
' Fingers extended 

Little and ring- fingers extended 
| Elbow flexed 


| Fingers flexed 


>», extended 
Elbow flexed 


Fingers extended 
Wrist adductec 


Fingers flexed 


eS extended 


Elbow flexed 


| Elbow flexed 


Thumb extended 
flexed 


” 


| Fingers ,, 
Wrist pronated 


Wrist supinated 


» Hexed 


Shoulder abducted 
Wrist extended 


Elbow flexed 


Wrist pronated 


| Elbow extended 


flexed 


” 


Elbow flexed 


Shoulder protracted 


and Gorilla 


151 


Srd movement. 4th movemen 


Elbow tlexed 


Shonlderretracted 


Shoulderadducted' 


Fingers flexed 


| 
Thumb adducted 


Thumb adducted | Fingers flexed 


Shoulder raised 


Thumb adducted 


Wrist flexed 


Wrist flexed 


Elbow flexed 


Numeral or 
letter indicat- 


ing response. 


152 


Leyton 


and Sherrington 


Appears in } | 
maps under 
anthropoid Ist movement. 2nd movement. 3rd movement. 4th movement. 
species, 
C, O, or G. 
Ce] Shoulder and elbow flexed simul- 
taneously 
c dropped from a raised post- 
ure which it had been 
maintaining | 
Cae Hallux adducted 
cio} - 33 Hip extended Ankle moved [xed 
Cc} oO; > extended Other toes extended Ankle dorsal- | Hip flexed 
(Ohl i 5 flexed 
CrOnlG :; flexed 
} G 5 “A Ankle flexed 
C ~ ss | Other toes extended 
C as ¥ 3 flexed Ankle extended 
C G A 35 Pe z flexed 
(0) G s,  adduected a extended 
|... | G pe iH flexed [an 
Cc} oO : Pe 5 33 a Ankle extended | Hip extended 
el O ms J ; Fe i | 5, flexed 
C G :, abducted 5 spread Knee flexed 
(G |G s,s  «adducted Ankle fiexed 
Seale Be aG :,  fiexed Other toes flexed 
Cars | G ;.  adducted 
ae ee ., abducted Knee extended 
OF 0) and all toes flexed 
COleee| er . ms wi Hip extended 
Cy Ou lee es 3 * 5 | Ankle plantar-flexed Knee flexed 
CamOniie: z ; 3 extended | 
ae | Fa Ge 3 = is 5 | Ankle dorsal-flexed Hallux abducted | u 
Cue: =e ¥ flexed {hase i is .» adducted | Hip extended 
is fs 53 Me 3 | Planta inverted eee 
.: | 2: op 6 by ' Ankle plantar-flexed Knee flexed Hip flexed 
C | 2nd toe flexed 
Oa ;, extended I 
C | .. , G | 2nd and 3rd toes flexed 
C/o Three outermost toes extended 
CPs Digits except hallux spread | 
OF 8) i Hos. ony s, extended 
C | G os 5 én = Hallux abducted Ankledorsal-tlexed 
C 3 e = a Ankle dorsal-flexed 
Cc 93 ; i | 5,  everted 
(opi : tlexed 
Ze G 2 i as 5 | Hallux flexed Hip extended Contralat.albdom. | 
ce ; | * wall contracted| 
C G Ap A | Ankle flexed Hallux adducted i 
CAN yer es 3 » x 5p i 5 Knee flexed } 
.. | O | .. | Hallux and toes flexed” dorsal-flexed s 33 Hip extended 
G G Digits except hallux flexed ~ x Hip flexed Anus everted 
Ca Ge 35 an = a - oe Anus everted 
(OTR ioc 4 nr a As "i 5 5D AA 
CaOr alles as . < * Hallux adducted Ankle dorsal-flexed) Hallux abducted 
-. | O-| @ | Two outermost toes flexed 3rd and 2nd toes flexed Hallux adducted 
Cc 3 e ~ x | if B 2 Ankle plantar- | Hallux adducted 
flexed 
C |... G | Hallux and toes spread widely | 
Ci (ORG 55 a flexed | Ankle plantar-flexed 
Cal LOaITG xi - ~ | Anus everted 
Cc; oO . Ankle flexed dorsally | 
C|/0|G * _ = | Knee flexed 
¥ * | Hallux and toes spread Knee tlexed 
C G 5 | 'Loes flexed Hallux adducted 
Cc as 35 extended 
c * flexed Knee flexed 
Cale ehiG = Ae Es Hallux abducted 
Coon i= oy % an Ankle rotated outward 
C!0);G :, DPlantar-flexed 
Cae Ge es ‘ _ Toes flexed except hallux Hallux adducted 
CUT OuEG. 5 i : Knee extended Hip extended 
Se ee ; an 35 Ankle adducted 
oo || 0) ; 3 a Hip extended 
(0; Be ; 55 i Hallux extended 
(e G 3 a > ; adducted 
50 || Yap. leae “a . ; | yn om ‘Toes tlexed 
CaOn} ’ A a | Toes flexed Knee flexed 
O i; 25 ae | Hallux fiexed 
ae >»  everted | 
Oa set sae ‘a 33 | Little toes extended 
Cel ss .» inverted Hallux adducted 


' 
== -_ ee __ 


The Excitable Cortex of the Chimpanzee, Orang-Utan, 


lst movement. 


Knee flexed 


» «extended: 


Hip flexed’ 


rotated inward 


7 ” 


> 
extended 
” 


a ” 


rotated outward 
abducted 
adducted 


rotated inward 
abducted 


” 
”? 


.. Flexed knee relaxed by inhibition 
. | Hip flexed 


‘Toes except hallux flexed 

| Conjugate deviation of eyes toward 

| contralateral side 

Contralateral eye turned toward con- 
tralateral side 

Conjugate deviation of eyes toward 
contralateral side 


” ” 2 ” 
Conjugate deviation of eyes toward 
contralateral side and depressed 
Conjugate deviation of eyes toward 
| contralateral side and slightly raised 
Conjugate deviation of eyes toward 
contralateral side 
Contralateral eye toward contralateral 
side and somewhat upward 
Conjugate deviation of eyes to contra- 
lateral side and somewhat upward 


. Ipsilateral eye turned toward contra- 


lateral side 
Upper eyelids of both eyes raised 
Lower eyelid of contralateral eye 
lowered 
Opening of both eyes 


” 39 
33 bed 
a: 7” 
3 ;, and converging of 


eyeballs 
.. With marked depres- 
sion of lower lids 


” 39 
| Contralateral eye closed 
Lowering of upper eyelid of contra- 
lateral eye 

Closing of both eyes, contralateral 
_ the more 

Raising of lower lid of contralateral eye 
Closing of both eyes, contralateral 


neo 

Ses Appears in 
#= & | naps under 
of anthropoid 
Ent Species, 
Fae ©, O, or G. 

—| 

$31 | C| Oo | G 
382 | C/..|.. 
338 |..| 0/|G 
334 ora ee 
335 pe | ae 
336 || C | G 
33 | C| O| 
$39 | C|..| G 
340 c|0/|G 
S41 Cc Oo G 
342 Cio; @G 
343 clo - 
344 Fetes 1G 
345 ot a ae 
346 Ci.. |G 
S47 te O: lis. 
ms }..|..| G 
349 mee ew o's 
350 eh... |G 
351 Gr 0:1 G 
352 OD AS) 
353 Bree Ts 
354 Os 
355 GLO}... 
356 “ee ee 
357 4 ee 
358 i G 
359 Ral oie 
360 eet 0 

361 As / a 
362 ee ae 

363 Cel 

364 LC ea 
370 c 0 G 
ae 

872 | C/0/|G 
3733 |c|0,/G 
S18 ee ? 
375 

376 | C 

377 | 

378 

379 C / 

: | 

380 C iG 
Sa 
382 Cc} 90 
383 «| GC G 
384 y hes’ 
385 | C {G 
386 C |G | 
387 | me 
— ae G 

a (0) G 

b fe G 

e € }. 0:1 G 

a 4 

= C 


the more 


2nd movement, 


Toes flexed 
Hip flexed 
., adducted 
extended 
Ankle flexed 
‘Toes flexed 


Hip extended 


| Hip adducted 
| _,,. Totated inward and adducted 
Hallux tlexed 
” extended 
Ankle flexed 
Knee flexed 
Hip extended 


Knee extended 
Contralateral wall of abdomen 


Knee flexed 
» extended 
| Ankle extended 


| 
| 


Contralateral wall of abdomen con- 
tracted 

| Knee extended 

| Contralateral wall of abdomen con- 
tracted 


Knee flexed, ankle plantar-flexed 


Ankle plantar-flexed 


Both eyes opened 


i) 


Pupils dilated 


Both eyes opened 


Both eyes turned contralaterally and 
downward 

Conjugate deviation of eyes to oppo- 
site side 

Face turned to contralateral side 

Eyeball turns to ipsilateral side 


Eyes turned to contralateral side 


33 


| 
| 


| Movement of jaw 


and Gorilla 


Srd movement. 


Ankle plantar- 
tlexed 


| Ankle flexed 


| Ankle extended 


| All digits flexed 


| 


| Toes except hal- 
Inux flexed 
' 


Neck turned to | 
contral. side | 


| 


Face turned to) 
opposite side 


Face turned to 
opposite side 


153 


ith movement. 


Hip tlexed 


| 
| 


Numeral or 


154 Leyton and Sherrington 
A ip = a ie a a, yal 
Betis 
S2 Appears in 
SS | maps under 
irl | anthropoid Ist movement. 2nd movement. 3rd movement. 
52 | species, 
ae | C, 0, or G. | 
f _ | Closing of both eyes, contralateral | Retraction of contralateral nostril Retraction con- 
the more : tralateral angle 
of mouth 
C | G , re , Fingers flexed 
om pales é “ a Index extended 
[bers Hei ; ; Screwing up of nose 
(0; | G _ Face turned to opposite side 
1c ; Drawing down of opposite eyebrow 
and forehead skin 
G , 2 Elevation of contralateral pinna 
ce | G 55 5 Retraction of . 
| | G Ae : Contralateral nostril raised Contral. forehead 
wrinkled 
* 5 Si ee ae pinna retracted Contral. angle of | 
Cc | Anus protruded chiefly on contra- mouth retracted 
lateral side 
» Contralateral side of abdomen con- 
tracted 
| 5 a 33 Hip flexed 
| | Raising’ of opposite ey elid 
3 and eyebrow 
=.) | Frowning on contralateral side Face turned to contralateral side 
c G | Contralateral nostril flattened Contralateral angle of mouth retracted; Tongue moved 
eet .. | Hyoid raised 
c | .. | G | Larynx raised (observed from outside) 
as in swallowing 
Cc | O | G | Movement of neck turning face to 
| contralateral side 
R | O | .. | Contralateral nostril lifted and opened 
s | O 75 ad Couvtralateral pinna moved 
d oO : 20 me ttt ittened Upper lip moved 
WwW | G Contralateral angle of mouth retracted 
oY, C | .. | G | Nostrils drawn down Mouth moved Eyes closed 
Z Gro | Eyes closed Face turned to 
opposite side 
P C | Contralateral pinna retracted 
Pe; oi) | a ; ; Eyes closed Eyebrow moved 
P, CNG 93 Lifting of contralateral eyebrow 
P, Ou! : 33 Contralateral angle of mouth retracted 
P, : | moved, upper 
part only 
Aordtece, han protracted 
Gallas | G | Vocal cords addueted 
C Js. | -- ;, With emission of 
: a sound 
Cree AG Ms especially the 
contralateral one 
asaitintete | abducted | 
oO | Pille u's of fauces bilaterally adducted | 
C0} % - e Tongue curled and heaped up at back 
of mouth 
oO . | neck turned face toward contralateral Contralateral pinna moved 
| side 
-: » Mouth moved 
Cc aS 53 Nose moved 
Cc Nea 55 of 3 5 | Eyes closed 
C G | Chest muscles of contralateral side | 
| contracted 
H » ” os .. | Neck turned to contralateral side 
C G Chest and scapular muscles of contra- 
lateral side acted (latissimus dorsi) 
Cc G | Lower part of contralateral side of 
| chest acted 
Cc G | Contralateral side of abdominal wall 
(om | Arching of back with action of contra-| 
Bi | | lateral rectus abdominis | 
Cc | G | Muscles of contralateral side of ab- 
| dominal wall, and contralateral 
| loin and anus } 
Cc | | Muscles of contralateral side of ab- | 
| dominal wall at umbilical level | 
Gal Muscles of contralateral side of lower | Anus protruded 
abdominal wall 
Cc | G |Musclesofcontralateral sideofandomen| Contralateral hip 
Cc | * 3 + Ee re L- Movement of 
| chest wall 
C | a) 53 3 Pe - ‘ _ Anus protruded 
Cc | | Retraction of contralateral shoulder | 


4th movement. 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 155 


Index to Primary Movements Listed. 


Numeral or letter in list. Numeral or letter in list. 
lips. ‘ i , : ; . 1-87, 39-58, 61, 99 ring Nnger . - . 162, 168-170 
nose. : 5 - ; SAB ARE vii Rp little finger ; : : 163, 170 
pinna of ear : ; ; : : P P, P, P, fingers without thum!) : . 171-181, 191 
cheek . ~ : ; . ‘ : 38, 110, 112, 113 fingersandthumh  . ; ; . 182-190, 192-194 
chin. . : F . si three ulnar fingers. . 195-200, 207 
eyebrow and frontalis - , : : DLE whole hand ; : ; . 202-206 
tongue . ; , . . 59, 60, 62-97, 99, 115 wrist. : : : : . 208-230 
jaw . J . 39, 43, 45, 46, 101 109, 111, 114 elbow . ; ; 231-287, 256 
hyoid region 3 : : f F cae shoulder. ; ; ‘ R . 288-256 
fauces . ‘ , : : P 5 - ; . Oo,” chest wall . : ; , ; . VI.-IX., 253 
palate . ‘ A . ; F ‘ ‘ . 98 abdominal wall . , . : X.-XVIII., 254 
uvula . . : : : , ; ‘ . 100 hallux . ; : ; ; ; P : . 260-285 
vocal cords : : .a, B, y, 5 2nd toe P > : : . 287-288 
larynx as observed from without — L 2nd and 3rd toes ; ; ‘ k ; : . 280 
eyelids, closing . - ; .a-h, k, m, n, Dp 4, ty w three fibulartoes ‘ ; . 290, 305 
eyelids, opening ; : ‘ “ ~ . 880-388 digits except hallux . ! é 91 - 290, 301-304, 364 
eyeball : * . : : ; P . 3870-379 all digits . : : ; . ‘ . 300, 307-3809 
neck. : . : ‘ : ‘ ; N, II.-V. | ankle . : ; : ; ; : . 810-330 
thumb ‘ ; ; . F - ‘ . 119-141 knee. , : ‘ ’ ; . 5831-389, 362 
index finger : . : : . 142-160, 158, 207 | hip ‘ ; ; ; ‘ , ; . 340-361, 363 
middle finger. 2 ; . 154, 155, 157, 161,162 | anus . : ‘ ‘ ; : i 3 Way AM ae: 


Buried Portion of the Motor Cortex. 

It will be seen from the accompanying charts and the foregoing list 
that a good deal of the excitable motor cortex lies tucked away from the 
free surface of the hemisphere, buried in the fissures adjoining gyrus 
centralis anterior (19). We have explored some of this buried portion of 
the motor area by faradisation. The unipolar method of stimulation is 
suited for such exploring better than is the bipolar; but the laying bare of 
the deep surface in a fissure necessitates destruction of one wall of it, and 
hemorrhage and interference with the minuter local circulation render 
difficult the successful examination of any large continuous length of a 
fissure in one and the same specimen. Our results have therefore been 
obtained piecemeal from a number of hemispheres. Fig. 6, A, B, illustrate 
the deep points localised in two such experiments ; fig. 6, A is a left hemi- 
sphere: fig. 6, B,a right, the map of which has been reversed for easier 
comparison with fig. 6, A, and with other figures. 

The results of our various experiments on this taken altogether revealed 
no movement from the buried motor cortex which was not elicitable at 
one time or another in one specimen or another froin the free surface of 
centralis anterior itself. The buried portion in sulcus centralis extended 
along the whole length of the anterior wall of that fissure except for its 
extreme upper tip, where motor cortex leaves the fissure and lies a little 
forward of it. In some places the motor cortex seems to pass down the 
whole depth of the anterior wall of the fissure, and not far below the inferior 
genu it seems in some individuals to occupy the deeper portion of the 
fissure’s posterior wall also. It seems to extend less deeply than elsewhere 
into the fissure at two places, one of these being the lower part of genu 
inferius, the other lower part of genu superius, the shallowing being more 
marked and sharper at the former. The former of the two corresponds 
approximately with the region for neck lying between arm area above and 
face area below. The latter corresponds with the region for abdominal 
wall and chest wall lying between arm area below and leg area above. 


156 Leyton and Sherrington 


And we have obtained movements of neck and trunk respectively by 
actual faradisation of the anterior wall of the fissure at those two places. 
The fissure being of considerable depth, exceeding 12 mm. in several 
places both in the chimpanzee and orang (we have not explored it in the 
gorilla), the amount of excitable area contributed by it to the motor tield 
of the cortex in the anthropoid is quite large. Fig. 15 (vide infra, p. 171) 
illustrates this. It is schematic, but not wholly so; it was prepared from 
a chimpanzee hemisphere, in which a number of points of the buried motor 
cortex were actually determined by faradisation, and the depth to which 
the fissure was excitable was tested ata series of places. With these as 


Fic, 6.—A, chimpanzee 11; left hemisphere, showing responses obtained at opened-up parts of 
some sulci. On the free face of the convolutions some of the responses evoked there are marked 
into the map to serve for orientation. bB, chimpanzee 15; right hemisphere reversed, re- 
sponses from opened-up parts of sulci and from free surface. The animal was of the variety 
Troglodytes calvus, and very intelligent. 


a basis, the rest of the deep contour is given by interpolating determina- 
tions obtained in other chimpanzee hemispheres. In our observations the 
posterior boundary of the motor cortex lying hidden in sulcus centralis 
seems to be more abruptly and sharply delimited than is the anterior 
margin of it, lying largely on the free surface of the hemisphere. 

The motor responses yielded from points buried in sulcus centralis 
corresponded for the most part rather closely with the motor responses 
yielded by the free surface of centralis anterior of about the same 
horizontal level. A good deal of the local area for pinna seems to lie 
buried in the sulcus close below inferior genu, and we have obtained pinna 
movements from the anterior wall of the sulcus at that place in specimens 
where we could not elicit them from the free surface of the gyrus. The 


? 


Ft 
261 392 299 39) SL) 4: 


PII LIS 3557 


- 


30 A320 318 3 
354i Ai 29025 35 4 ‘ 
20 317 2) 


Fic. 7.—A and B, Orang 1, left hemisphere ; three perspective plans. The numerals and letters 
refer to the ‘‘ List of Responses,” pp, 148-154. 


158 Leyton and Sherrington 


precentral sulci, superior and inferior, also contain portions of the motor 
cortex. ‘These sulci are far more variable in their extent and position in 
the anthropoid than is suleus centralis,so that it is not easy to make a 
general statement as to the amount of motor cortex they contain that can 
apply strictly to all cases. In the fig. 15, from a chimpanzee, is represented 
the amount buried in them in that specimen as experimentally determined. 
Moreover, the determination of the exact position of the anterior limit of 
the motor cortex is even on the free surface a matter of some artificiality, 
because in the anterior direction the motor field as examined by faradisa- 
tion seems to fade off graduatim, so that a prolonged series of stimulations 
in that neighbourhood produces, by inducing “ facilitation,” a limit set 
farther forward than under a brief decisive examination by faradisation 
at a restricted number of selected points. 

In his “Localisation of Cerebral Function” Campbell (8) has fur- 
nished an admirable and full account of the structural types of cerebral 
cortex and their topographical distribution not only in man but in the 
chimpanzee and orang. It is instructive, therefore, to compare the limits 
of the motor field as determined by faradisation in those anthropoids with 
his “precentral area” determined by cell and fibre lamination. The 
posterior borders of the two as delimited by these two different methods 
seem to agree so closely, that there can be little doubt that as regards that 
limit the two fields or areas are the same. In regard to the anterior 
border, the motor fields boundary seems to lie, especially in its lower two- 
thirds, farther forward than does that of Campbell’s precentral area. The 
anterior boundary, as determined by faradisation, is, however, not a sharp 
one, and its situation seems to vary somewhat from specimen to specimen, 
As placed by us, it certainly appears to lie for the most part in the inter- 
mediate precentral area of Campbell. Opposite the “arm area” it lies 
not far behind the anterior boundary of the “intermediate precentral area,” 
but opposite the “leg area” more considerably so. Opposite the “face 
area” it lies very much farther behind the anterior limit of “inter- 
mediate precentral area,” although in front of anterior limit of the pure 
“ precentral area” of Campbell. Campbell in his original description 
furnishes a number of arguments in favour of his “intermediate precentral 
type” of cortex possessing motorial functions, though differmg from the 
precentral type or motor cortex pure. The gradual shading off of the pure 
motor field in the anterior direction, as experienced in our observations, 
and the variability of its anterior edge, as mentioned above when faradised 
under different experimental conditions, seem to us to lend support.to his 
contention, although the latter is put forward on other evidence. 

On the whole, we should estimate that in the anthropoid brain the 
portion of the motor region which lies buried in the sulcus centralis and 
other fissures amounts to not less than about 35 per cent. of the whole 
motor region. 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 159 


Remarks on the Grouping of the Responses 
of the Motor Cortex. 


An occurrence met in some of our experiments was that in the course 
of examination of gyrus centralis anterior some small area of its surface 
might exist whence the faradisation failed to evoke responses. An in- 
stance is figured in fig. 2, B, close to the genu inferius of sulcus centralis. 
The appearance of the cortex at such a place would reveal to inspection no 
obvious circulatory disturbance; nor so far as we were aware had any 
damage been inflicted there. But in many experiments the whole motor 


Sil 
333 


Fic, 8.—Orang 2. A, left hemisphere; B, right hemisphere reversed for comparison with A, 
Some only of the responses obtained are entered on the map. 


field systematically explored revealed no obvious gap in it. Beevor and 
Horsley in the brain of the orang they stimulated met with relatively 
large and numerous gaps of this kind, and supposed them characteristic 
of the motor area of the anthropoid brain. Franz records meeting with 
small areas not yielding responses in the motor field of the macaque 
monkey. In our experience, a return later in the experiment to the small 
area which had not yielded responses found it still unyielding of response. 
Such an area was on several occasions ascertained to have no counterpart 
that we detected in the hemisphere of the opposite side. Nor were the 
places of occurrence of such seemingly non-stimulable gaps the same in 
hemispheres of different individuals, a finding in conformity with that of 
Franz in macacus. In our experience, such a gap was perhaps less infre- 


quent than elsewhere about the region where face area meets arm area. 
VOL. XI., NO. 2.—1917. 11 


160 Leyton and Sherrington 


It may be noted that among the chimpanzees we examined were two 
very young ones, the younger of them, though in good nutritive con- 
dition, weighing only 2°240 kilog. In one of these we found a cortical 
differentiation of the finger movements at least as great as in any other of 
the anthropoid brains we explored. In this animal we obtained from 
appropriate points in the cortex isolated movements of the little finger, 
both isolated flexion and isolated extension; also isolated movement of 
the 2nd toe, both of flexion and of extension; also movement of the 2nd 
and 3rd toes without movement of the other toes. The animal was so 
young as to be infantile; it was fed from a sucking-bottle, and had the 
petulance and habits and cries of a very young animal. 

Epilepsy.—Prolonging the faradisation, especially strong faradisation, 
of a spot in the motor surface usually induces not only a considerable 
“march” or sequence of responsive movement, but also, as is well known, 
an epileptiform convulsion. Our experiments were not directed toward 
observation of these, but we induced such effects from time to time. We 
found them easily provoked in the anthropoid, but not obviously more 
readily than in small monkeys such as macacus and calothrix. A differ- 
ence in the two cases seemed the greater relative ease with which in the 
anthropoid an epileptiform convulsion could be evoked in this or that 
small region of musculature without the convulsion spreading beyond that 
part. Thus it could be evoked in the index finger, in the angle of the 
mouth, or in the toes, and remain confined to the field in which it started : 
such restriction is, in our experience, quite uncommon in macacus or 
calothrix. 

“Epilepsy ” was evoked readily in the “baby” chimpanzees coming 
under observation; it seemed neither more nor less readily obtained in 
them than in the grown specimens. On the other hand, the ease with 
which it was evoked, and the tendency for it to occur in the course of an 
experiment, appeared to us to vary distinctly in different individuals; in 
some individuals stimulation of duration and intensity too small to evoke 
it usually, tended to evoke it from the very beginning of the experiment, 
and that tendency continued throughout the experiment. 

It may be of interest to remark that in ablation experiments with 
small monkeys we have sometimes found a collodion dressing applied to 
the scalp over the removed area of bone produce severe epileptiform 
convulsions, which ceased at once on removal of the dressing. The 
shrinkage of the collodion in such cases caused the dressing to press upon 
the scalp and underlying brain, the surface of the dressing over the 
removed piece of skull becoming flat or slightly concave outward. 

A few general remarks may be offered in regard to certain of the 
movements evoked and the representation of separate motile parts in the 
cortex. We follow for convenience the order taken in the foregoing index 
to the motor responses listed. 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 161 


1. Face Area. 


Face area so-called might be better termed head area, since it includes 
not only the face but tongue, palate and fauces, and larynx. Its upper 
boundary is usually with close accuracy marked by the level of the genu 
inferius of sulcus post-centralis. In some chimpanzees and gorillas, and 
especially, in our experience, in the orang, there is a tendency to the 
appearance of a third genu of the fissure below the ordinary genu inferius, 
And this third genu indicates approximately the level of subdivision of the 


Fic. 9.—Gorilla1. A, left hemisphere; B, right hemisphere reversed for comparison with A. 


so-called face area into an upper part in which movements of face proper 
predominate, and a lower part in which are represented tongue movements 
and movements of fauces, vocal cords, and palate. This third genu might 
be called a labio-lingual genu (fig. 7A), because at it the area where lip 
movements predominate as primary responses meets the area where tongue 
movements as primary predominate. In the upper part of the face area 
the movements elicited can bear for the most part an interpretation as 
being partial movements in mimetic acts. In the lower part of the face 
area the movements suggest for the most part their being parts of acts 
subserving feeding, e. g. chewing, mastication, deglutition, ete. 


162 Leyton and Sherrington 


Lips.— The upper lip movements are rather closely associated with 
movements of the nose, but the lip has a much wider focal field than has. 
the nose. The lips are represented largely together, but independent 
movements of both upper and lower lips were seen. The field of repre- 
sentation of the lower lip seems somewhat larger than that of the upper. 
The areas for the lips are much commingled, but the representation of 


ANUS 


Fie. 10.—Gorilla 1 ; perspective view traced from a photograph ; responses grouped 
diagrammatically. C, opposite the end of the sulcus centralis. 


the lower lip seems to extend or to have its chief seat rather lower down 
the centralis anterior than does the upper. Pouting of the lips was 
distinct and not uncommon. Retraction of contralateral angle of the 
mouth, either primary or secondary, seemed the most common of all the 
lip movements. 

Nose.—Movements of the nose seemed better developed, and to have 
a wider cortical representation in the orang than in chimpanzee or gorilla. 
They were not so marked in the orang, however, as they were found to be 
in the baboon (36, 7). 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 163 


Pinna of Ear.— Movements rarely alone, almost always associated 
with other movements of the face; focal area lies partly buried in sulcus 
post-centralis. The area seems larger in the chimpanzee than in the 
orang. 

Cheek and Chin.—Movements were elicitable, but not common. 

Eyebrow and Frontalis.—Movement always contralateral; field in 
upper part of facial area. 

Tongue.—Movements of tongue were obtainable from a very large 
area, in which they were usually the predominant primary movements. 
They were extremely varied in their form and sequence, almost bafHing 
verbal description. For the most part they could be grouped under the 
headings retraction, protrusion, rolling on long axis, upeurving of base or 
tip, and hollowing of upper surface from side to side. 

Protrusion very rarely carried the tongue tip beyond the lips. The 
appearance of the movements frequently suggested that they were part 
actions in mastication, licking, lapping, and swallowing. Thus one not 
infrequent was a thrusting of the tongue against the inside of the cheek- 
pouch as though to remove food thence; again, a rhythmic movement of 
licking or lapping; again, a heaping of the back of the dorsum against the 
back of the palate, followed by contraction of the faucial opening as though 
in swallowing. Occasionally the tongue was drawn back or thrust forward 
straight; much more commonly the retraction or protrusion was deviated, 
the deviation being sometimes to the ipsilateral, sometimes to the contra- 
lateral side. Retraction and protrusion were evidently much commingled 
in their representation in the cortex, but on the whole protrusion seemed 
situated lower down the convolution than was retraction. Sometimes the 
protrusion of tongue was accompanied by closing of jaw, and then 
occasionally the tongue was nipped by the teeth, recalling the biting of 
the tongue in epilepsy. On many occasions the points of excitable cortex 
farthest down of all in the convolution evoked movement confined to 
the tongue tip. 

Jaw.— Opening and closing were both elicitable, but the former has a 
considerably larger field of points than has the latter; the latter's field 
seems to lie the farther forward and not to extend nearly so far down, 
at least as a primary movement, although in sequence to opening it extends 
far downwards. Rhythmic chewing, a movement observed by Ferrier as 
readily elicitable from the cortex of the cat, dog, and monkey, was observ- 
able in the anthropoids, and was got from points low down and far forward 
at the foot of the convolution. The jaw was not infrequently deviated 
towards the contralateral side as well as opened or closed. By dividing 
the symphysis it was found that the cortical representation is mainly uni- 
lateral, although when the two lateral halves are normally conjoined by the 
symphysis the unilateral representation in the hemisphere is mechanically 
obscured. 

Hyoid.—Lifting of hyoid from a restricted part of lingual area. 


164. Leyton and Sherrington 


Fauces. — Movement usually bilaterally symmetrical in appearance ; 
generally from posterior part of lingual region about half-way down and 
from a quite restricted region often in association with heaping up of 
tongue at the back of the mouth. 


Fic. 11.—Gorilla 1. 


Vocal Cords.—Movement almost always adduction, bilateral, but 
sometimes clearly more marked on contralateral side. Focal field small 
in the anterior and lower part of the face area, i.e. adjunct to the 
lingual field. 

Movements of the Eyelids and Eyeballs.—Cortical stimulation 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 165 


draws a sharp distinction between eyelid movements of closure and of 
opening respectively. This is the more striking, because movements of 
opposite sense implicating one and the same part, e.g. thumb, index, ankle, 
wrist, tongue, lips, even elbow and jaw, are not easily or even commonly 
separable in the cortex by reason of their foci of representation lying con- 
siderably remote one from the other. But the tields for eye-opening and 
eye-closure respectively do lie considerably separated apart. 

The great field of excitable cortex which lies open to examination in 
the free surface of gyrus precentralis may be termed the precentralis motor 
field; and we may include under that term the whole of the seemingly 
continuous field of motor points which occupies as well as the free face of 
gyrus precentralis the adjoining portions of sulcus centralis and of sulci 
precentrales and parts of the free faces of the gyri annectantes connecting 
gyrus precentralis with the frontal convolutions. Among all the numerous 
and varied movements which faradic stimulation applied at the appropriate 
points evokes from this great field, opening of the eye does not appear to 
be included, neither do movements of the eyeball. But closure of the eye 
is well and definitely included among the movements elicitable from 
precentralis field. 

Closure of Eye.—The place in that field which yields eye-closure lies at 
the level of, and extends a little above, and to a wider extent below, genu 
inferius of sule. centralis. It meets, as examined by the electrode, 
the lowest points of hand area (thumb) above; it is intimately adjunct to 
areas for ear, nostril, neck, and lip, occupying part of the upper portion of 
face area, and is traceable with the electrode into tongue area. The eye- 
closure is obtainable with faradic stimuli of the same strength as suffice 
for other motor responses from precentralis and with the same readiness. 
The movement may be (e.g. with weak stimuli) restricted to closure of 
opposite eye only, or even to isolated movement of the upper or lower lid 
only of that eye. With moderate stimuli the closure is of both eyes, but 
practically always is more vigorous in the opposite eye. The closure 
sometimes has the appearance when the animal is not too deeply narcotised 
of being executed against the animal’s will, for it occurs while the other 
eye remains almost open, and on withdrawal of the cortical faradisation 
the contralateral eye, as also the less closed ipsilateral, re-opens again 
immediately and quickly. 

Opening of the Eye.—This movement is observable under stimu- 
lation of the cortex in various widely separated regions. It may occur, so 
to say, in a desultory manner, and, in our experience, is prone to crop 
up unexpectedly. But in two regions it occurs fairly regularly; these 
regions are a frontal area anterior to the lower half of precentralis gyrus, 
and an occipital region including the calcarine area and the occipital pole. 

Taking the second region first, the opening of eyes elicitable thence is 
clearly associated with a turning movement of the eyeballs toward the 
opposite side. The eye-opening, like the eye-turning which it accompanies 


166 Leyton and Sherrington 


(36) is elicitable from this region much less easily and regularly than are 
the ordinary motor responses evocable from the precentralis motor field. 
Moreover, the points which here yield it seem, as tested by the electrode, 
to lie in a scattered manner, not constituting a continuous field of excit- 
able points. Examination of the region shows that reacting points are 
most numerous along the area bordering the posterior part of calcarine 
fissure, and therefore on the mesial face of the hemisphere, but both in 
chimpanzee and gorilla the response was obtained also from a few points 
of the lateral face of the hemisphere at its occipital pole; the experiments 
giving this were on quite young animals, except in the case of one adole- 
scent chimpanzee. 

The other region whence eye-opening is elicitable, the frontal, is a large 
one. It embraces a considerable part of the 2nd and 3rd frontal con- 
volutions, and seems separated from the “ precentralis motor region” by 
an intervening strip of “silent” cortex, although this strip is sometimes 
encroached on almost to extinction. Elicitation of eye-opening from this 
region, like its elicitation from the occipital field, though apparently in a 
less degree, is irregular, and requires stronger faradic currents than are 
required for exciting motor responses from precentralis region. The move- 
ment when evoked has commonly a more deliberate execution, and the 
points which yield it are in any one experiment scattered in discrete fashion, 
instead of forming a seemingly continuous excitable field—as obtains, 
for instance, with the points yielding eye-closure in precentralis. The 
movement is practically always bilateral, often without obvious trace of 
preponderance of vigour for the contralateral eye. With it is associated 
turning of the eyeballs; almost always conjugately away from the side 
stimulated, occasionally however convergently, and then sometimes toward 
a plane continuous with sagittal plane of head. Sometimes the eye-opening 
precedes the turning of the eyes, sometimes it follows it. In our experience 
the former is more frequently the case with the lower part, e.g. 3rd frontal 
gyrus, of the frontal region, the latter with the upper part of the region. 
The opening of eyes tends to be followed, especially after reiterated stimula- 
tion, not only by eyeball-turning, but by turning of the neck and head in 
addition. This secondary and tertiary movement is always directed so as 
to turn the face away from the side stimulated. 

In some experiments, e.g. young gorilla (fig. 12), the frontal area yield- 
ing eye-opening seemed to be subdivided into two by an inexcitable strip 
running horizontally across it. But considering the scattered distribution 
of the points in this field, this subdivision may be one that more extensive 
experimentation would break down, and certainly in some specimens it did 
not seem confirmed. Where it occurred the lower of the two sub-fields 
usually yielded eye-opening precedent to eye-turning, and the upper sub- 
field eye-turning precedent to eye-opening, and not rarely altogether 
without the latter. 

It was said above that the movement of eye-opening seems absent from 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 167 


Fic. 12.—Gorilla 2, very young. A, left hemisphere; B, right hemisphere. The extent of the 
excitable area indicated by stippling, and its gradual merging forward into inexcitable indi- 
cated by decrease in density of stippling. Only some of the responses are mapped. The areas 
enclosed in the heavier dotted lines indicate the portions ablated (v. infra, p. 200). 


Fic. 13.—Gorilla 3, young. Perspective view of right hemisphere, traced from a photograph 
kindly made by Dr W. A. Campbell. Reversed for ease of comparison with fig. 10. Only 
some of the responses obtained are mapped. 


168 Leyton and Sherrington 


the long list of motor items assembled in the large precentralis motor field. 
Although that, as a broad statement, is true, it requires some modification, 
inasmuch as occasionally, and, in our experience, very rarely with weak and 
moderate stimuli, eye-opening is evoked from precentralis. It is not then 
a primary movement, in our experience, but is secondary to, or developed 
from, the turning of neck and head elicitable from a restricted area between 
hand and face regions; and with it sometimes occurs movement of eyeballs 
to opposite side, also not a primary movement from precentralis. More 
usually this neck movement, which is a movement carrying the face away 
from the side stimulated, is associated with movement of eye-closure, as 
mentioned above. From the evidence of our experiments in anthropoids, we 
infer that the eye-opening sometimes elicited from this part of precentralis 
region is not to be taken as evidence of the existence in precentralis of 
motor foci there situate and directly executive of eye-opening, but rather 
of secondary connections of the precentralis neck focus with foci for eye- 
opening situate extrinsic to precentralis motor region proper. 

In addition to the instances of eye-opening coming under one or other 
of the three groups in the above category, there are instances of its occur- 
rence under faradic stimulation of still other regions of cortex. Such 
instances as these latter are those we had especially in mind in the opening 
paragraph as desultory, unexpected, and unreliable of repetition even at one 
and the same period of an experiment. When they have occurred they 
have been noted by us, and the places of their occurrence have variously 
included points in the first temporalis, calloso-marginalis, and post-centralis, 
as well as various parts of precentralis. In regard to the whole of this 
group of “desultory ” eye-opening responses, it is to be borne in mind that 
the movement of eyes-opening is one commonly accompanying an awaken- 
ing from sleep, and that the grade of narcosis under which the faradic 
examination of the cortex has to be carried out is one which in its depth 
somewhat resembles natural sleep. Any stimulus which arouses the animal 
is likely to evoke an opening of the eyes. For instance, the application of 
the faradic stimulus to the dura mater instead of to the cortex commonly 
does so. The eyes-opening has therefore to be accepted with much caution 
as evidence that the stimulus which evokes it is one really playing in a 
direct manner upon a “ motor” eye-opening centre in the cortex. 

Movement of Eyeballs.—This, as obtained from cortex cerebri in the 
anthropoids, is, as in the small monkeys, almost always lateral conjugate 
deviation to the side away from the stimulus. It is obtainable from (1) 
the calcarine region and occipital pole in the same area as that already 
mentioned under eye-opening; (2) a frontal area embracing a considerable 
part of the surface of the 2nd and 3rd frontal gyri, corresponding fairly 
well with the frontal area above mentioned under eye-opening. The con- 
jugate deviation of the eyeballs to the opposite side seems usually purely 
lateral, but sometimes the deviation is partly downward or partly upward 
as well as lateral; a partly downward deviation has, in our experience, 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 169 


been more common than a partly upward one. ‘To evoke these eyeball 
movements, whether purely lateral or not, requires, in our experience, 
stronger stimuli than are required for motor responses from precentralis 
motor area, and the responses even under these stronger stimuli are not 80 
regularly obtainable as are the responses from precentralis area (33). Nor 
do the points yielding them form, in our experience, a seemingly continuous 
field of excitable surface either in the occipital or in the frontal regions. 
The movements when obtained have further a slow deliberate development 
(39) usually, distinguishing them somewhat from the limb, face, and eye- 


Fic. 14.—Examples of ‘‘deviation of response” from a chimpanzee 
hemisphere, chimpanzee 19. Some only of the responses obtained 
are mapped, Left hemisphere. 


lid-closure movements evocable from precentralis region. The movement 
is usually bilaterally symmetrical, but not rarely the ipsilateral eyeball 
lags somewhat behind the contralateral. Very occasionally we have seen 
convergence of the eyeballs occur, sometimes very markedly, and as 
though to fixate a point approximately in a plane continuous with the 
sagittal plane of the head. After the lateral conjugate deviation has 
been obtained, it has been usual for the eyes to remain for some time in 
the posture thus assumed, and to return very slowly toward the primary 
straight-forward posture after the stimulus has been withdrawn. 

Quite exceptionally we have seen movement of the eyeballs produced by 
stimulation of the precentralis motor field. The movement has not been 
primary ; it has accompanied turning of the neck, carrying the face to the 
opposite side, and the region which has yielded it has been that of genu 


170 Leyton and Sherrington 


inferius, which contains representation of the neck. The eyeball move- 
ment has always been conjugate deviation of both eyes to the opposite 
side. It has, in our experience, almost always been accompanied by eyes- 
opening. Our inference from our experience of it is, that in the anthropoid 
cortex there is no focus in the precentralis motor field which represents 
eyeball movements in the same relatively direct way as do foci therein 
represent movements such as those of hand, face, neck, etc., regularly 
elicitable from the precentralia. We regard the eyeball-turning movement 
occasionally elicitable from the inferior genual portion of precentralis as 
secondary to the neck-turning foci, in the same way as we regard the eye- 
opening elicitable from the same portion as secondarily associated with the 
neck-turniny foci. 

As regards the eyeball-turning movement obtainable from the frontal 
region, this may be unaccompanied by eye-opening, especially so in the 
upper part of the region, in our experience. The eyeball-turning is easily 
detectable although the lids remain closed, the movement of the balls being 
obvious under the shut lids. 

We are disposed to regard the neck-turning elicitable from precentralis 
motor region as a protective movement mainly associated with closure of 
the eyes. The neck-turning movement elicitable from frontal region and 
from occipital region seems connected with the management of the direc- 
tion of the gaze. 


2. Neck Area. 


The area in which neck movements as primary or isolated responses 
are elicitable is, in our experience, small. The movements are closely 
associated with that of closure of opposite eye, and the movement is almost 
invariably one which turns the face away from the side to which the 
stimulated hemisphere belongs. We have not met with indubitable “re- 
traction” of the neck, although in an orang in which tetanus had been 
induced by inoculation with tetanus toxin, H. E. Roaf (386) and one of us 
observed “retraction,” which suggests that the observed retraction in that 
case was a deviated response due to the disease, and probably symptomatic 
of tetanus. To the small neck field lying on the free surface of the cen- 
tralis anterior, our observations show that, at least in some specimens, a 
part of the cortex buried in sulcus post-centralis has to be added. Neck 
field lies between arm area and face area, and seems to mingle more with 
the latter than with the former. Occasionally the turning of neck is to- 
wards ipsilateral side. 


3. Arm Area. 


Thumb.—Movements of thumb are among the movements obtained 
from the lowest part of arm area, and usually the predominant ones there. 
Their field abuts on, but mingles relatively little with, eyelids, neck, and 
“pinna of ear” fields. 

Index Finger.—The field for primary movements of this digit appeared 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 171 


in some of our specimens to be larger even than that for thumb. Exten- 
sion was a much more common movement than flexion, but isolated flexion 
was sometimes evoked. Extension of index as an isolated movement with- 
out any motion in thumb or other fingers was on very many occasions 
readily obtained. A not infrequent response from cortex when the resting 
posture happened to be one with adducted thumb and semi-flexed index 
was simultaneous abduction of thumb and straightening of index as if to 


Fic. 15.—Diagrams to illustrate the size and shape of the ‘‘motor” cortex of the 
chimpanzee as determined by faradic stimulation. A, shows the extent as observed 
on the free surface; B, shows the extent and shape as observed when the portions 
buried in the sulci are added to that on free surface. The line running through B 
marks the position of the sulcus centralis; all the shaded area behind that line 
represents the part of the ‘‘motor” surface buried in that fissure. The stippling 
to left of the shaded area in each diagram indicates the gradual fading off of excitable 
area in the anterior direction, making the actual demarcation of the motor field in 
that direction a somewhat arbitrary one. The smallest squares on the map represent 
mm.” of actual cortex surface in the specimen mapped. 


let go an object that had been picked up. Occasionally isolated extension 
of the terminal and 2nd phalanx only of index was evoked. In some 
chimpanzees we noted the using the singly extended index finger for 
various purposes to be habitual; thus, for picking the teeth after eating, 
for getting up a rice or maize-grain between boards of the cage floor. 
The marked individuality of this finger’s representation in cortex stands 
in harmony with such habituation. As an occasional response from cortex 
we saw extension of index finger accompanied by flexion of the three other 
fingers, no movement at all occurring in thumb. 

Other Fingers.—Isolated movements of other individual fingers were, 
as might be expected, much less elicitable. Isolated flexion of middle finger 


Leyton and Sherrington 


172 


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173 


zee, Orang-Utan, and Gorilla 


table Cortex of the Chimpan 


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174 Leyton and Sherrington 


alone was, however, obtainable occasionally, and notably in two very young 
chimpanzees. Isolated movement of annulus was never obtained. Closure 
of the whole hand was usually easily obtained, but the degree of separate- 
ness of the representation of the fingers as a group from that of thumb 
was marked. Over and over again all the fingers were extended or flexed 
without accompanying movement of thumb. 

Wrist.—The motor field for wrist is extensive, and both flexion 
and extension are readily elicitable, the foci for the two lying not 
together, although near one another. The responses of wrist are closely 
bound up with those of fingers, but the former's focal field lies higher up 
the convolution. Wrist responses sometimes were obtainable from points 
very far forward, in front of precentral sulci, e.g. fig. 6, and then commonly 
in association with index and thumb; but there also the wrist tended to 
have its representation higher upon the face of the hemisphere than either 
thumb or index. There appears no great predominance in representation 
of flexion over extension or extension over flexion in regard to wrist when 
observations made in a number of hemispheres are taken together, although 
in a single experiment on a single hemisphere one or the other may appear 
to predominate. 

Elbow.—Elbow has a large focal field, situate higher up centralis 
anterior than is wrist’s, and below the shoulder’s. Flexion of elbow 
predominates over extension in its representation. The two focal fields 
are commingled, but the smaller extension focus, in our experience, lies 
posterior to that for flexion; some of it lies buried in sulcus centralis. 

Shoulder—These movements are, as was to be expected, represented in 
a wide focal area, occupying their well-known position at the top of arm 
area. Their area extends into both central and precentral sulci. Genu 
superius varies much in prominence, and not rarely a small spur fissure, 
generally cutting into centralis anterior from behind, but sometimes from 
in front, lies partly, or rarely wholly, across the convolution at level of the 
genu. Into the lower wall of this spur fissure shoulder area sometimes 
dips. Shoulder area merges somewhat gradually into elbow area below 
and into a chest-wall area above. 


4. Trunk Area. 

Chest Wall.—There is a small area focal for movements of the 
contralateral chest wall. This les opposite or close below genu superius 
of suleus centralis. It lies partly buried in anterior wall of that sulcus, 
and in the spur fissure usually when that is present. It merges upward 
quickly into an area focal for movements of the abdominal wall. 

Abdominal Wall.—Movements of the contralateral abdominal wall are 
very regularly elicitable from a small area situate at the genu superius 
level. ‘The area merges in chest-wall area below and hip area above. The 
movements are, in our experience, always unilateral and contralateral. 
Some of the area commonly les buried in sulcus centralis, and in the spur 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 175 


fissure when that is present. Over the area one or more large veins 
commonly traverse the face of the convolution (fig. 18), rendering the 
experimental examination of the area somewhat difficult. From the 
abdominal wall area anal movement is sometimes elicitable secondary to 
movements of the abdominal wall. 


5. Leg Area. 


Leg Area.—The movements of the several parts of the limb seem more 
commingled in this area than are those of the separate limb parts in the 


ip wana 


/ 


ea ' r 
ay ® P 
Nar \entd / : 


en + Noes, 


rh, =e Fi % 
Dene dhulilelses \ *s Aonque 
echt ‘ : *. Palele Te. 
Wg f ‘ Ain ot Ra 
. r Mouth 
\Voow’ a) 
\waners 


Fic. 18.—The veins passing over trunk area of motor cortex in a large chimpanzee ; 
drawn by Professor Harvey Cushing. 


arm area. Nevertheless a general sequence of foci of main representation 
is recognisable; as the area is examined from below upwards this sequence 
runs hip, knee, ankle, and digits. The leg area extends over the mesial 
border of the hemisphere, and dips into the mesial surface for about one- 
third of the depth toward corpus callosum (figs. 2, i) By 9).10,33) 1415 1 
24). The area does not usually follow sulcus centralis to the extreme end 
of the sulcus, but leaves it a few millimetres below that, and slants obliquely 
forward over the mesial edge of the hemisphere. 

Hip.—The movements have a wide focal field; in our experience, the 
lowest situate of the movements is flexion. On the whole, extension of hip 
lies farther anterior than does flexion. In some specimens, notably in one 
orang, extension of hip was represented as a primary movement over a 


much wider field than was flexion, but the reverse is more usual. 
VOL. XI., NO, 2.—1917. 12 


176 Leyton and Sherrington 


Knee.—Extension, of knee as the sole movement of a response is rare, 
much more so than is flexion. 

Ankle.—Movement of ankle occurs often as a leading movement, but 
tends to be rapidly followed by movement of some other part of the limb. 

Digits.—Isolated movement of individual digits is not uncommon, as 
the “list of movements” shows. Some of the movements obtained from 
cortical stimulation of the anthropoid are such as we find difficult of 
execution ourselves. This was notably so with the foot area. Flexion of 
digits along with dorsal flexion of ankle we observed under cortical stimu- 
lation both in chimpanzee and orang. Extension of the 2nd toe isolatedly 
from the other toes was also seen. 


6. Perineum Area. 


Anal movement, usually protrusion, was elicitable fairly regularly and 
readily from a small area near the mesial border of the hemisphere in the 
anterior part of leg area, and apparently surrounded by this latter. The 
movement often seemed bilaterally symmetrical, but with weak stimuli was 
usually quite clearly unilateral and contralateral. Associated with it secon- 
darily was movement of abdominal wall, as has been noted in the smaller 


monkeys by Schafer (36) and by Jolly and Simpson (22). 


Inferences regarding Functions of the Motor Cortex. 


Franz (17) has recently obtained experimental evidence indicating that 
the functional topography of the motor cortex exhibits in Macacus rhesus 
demonstrable variation from individual to individual. The larger scale on 
which the motor cortex presents itself in the anthropoid, and the greater 
degree to which isolated movements of separate motor parts are elicitable 
from it, favours examination of the question, although our observations 
were not specially directed toward it when they were made. Compared 
one with another, the charts obtained from our anthropoid specimens of 
the same species exhibit, as said above, differences in detail; the amount 
of difference varies very greatly, as reference to those of the charts re- 
produced will show. The differences are present even in those brains 
in which the convolutional pattern is less dissimilar than usual, and are 
then for that reason better recognisable. One difficulty for such com- 
parisons is the fluctuating character of the sulci as landmarks for evaluat- 
ing the topography. Since we must suppose that the sulci have some 
functional significance, this fluctuation may itself be taken as an indication 
of individual variation of function. And certainly variation of convolu- 
tional pattern from individual to individual is, in our experience, one of the 
most salient structural features of the anthropoid cerebrum. Another 
difficulty in making the comparison from individual to individual is the 
fact, illustrated above, that the cortical motor points, or many of them, 
are within limits functionally unstable. The chart obtained from a motor 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 177 


region examined at one time and by one series of stimulations may not 
agree in detail with that obtained from the same motor region at another 
time and under another series of stimulations. But the differences between 
the charts from different individuals in our experiments seem too wide in 
most instances to be accounted for by merely temporal fluctuations of 
response, such as localisation experiments somewhat differently conducted 
might evoke from one and the same hemisphere. Inspection of the charts 
reproduced exhibits the scale of difference observed better than can verbal 
description. We regard them as indicating that individual variation of the 
functional topography of the motor cortex, as found by Franz (17) in 
Macacus rhesus, is demonstrable in the anthropoid species examined by 
us, and in at least as liberal measure. 

The list of motor responses taken as a whole shows that a very con- 
siderable number of different movements are obtainable from the motor 
cortex of the anthropoid, far more than can be obtained from the dog or 
macaque. Although of these a very large proportion may crop up in any 
single systematically conducted point-to-point examination of a single 
hemisphere, many of them do not. From our experience we imagine that 
had our experiments extended to a larger number of hemispheres, the list, 
which continually slowly grew in length as our experiments proceeded, 
might have grown a great deal farther. Another point obvious from the 
bare memoranda in the list, but still more obvious to inspection of the 
movements as they occurred at the time, is that the individual movements, 
elicited by somewhat minutely localised stimulations, are, broadly speaking, 
fractional, in the sense that each, though co-ordinately executed, forms, so 
to say, but a unitary part of some more complex act, that would, to attain 
its purpose, involve combination of that unitary movement with others to 
make up a useful whole. In evidence of this “fractional” character it is 
only necessary to note the predominantly unilateral character, as elicited 
from the cortex, of movements that under natural circumstances are 
symmetrically bilateral. Thus under cortical stimulation even such move- 
ments as contraction of the fauces, adduction of the vocal cords, closing 
and opening of the jaws, protrusion of anus, were often, indeed usually, 
detectibly asymmetrical, the execution being chiefly or wholly in some 
cases by the muscles of the contralateral side. A further point evident 
from the list is the considerable variety of combination into which these 
fractional movements were welded in the movement-sequences noted. Our 
main purpose being “localisation” of the primary movement, we did not 
usually, by pressing and prolonging any single stimulation, develop these 
sequences in our observations. Had that been done, the listed variety of 
them would doubtless have been greatly increased. Their variety, how- 
ever, even in the list obtained, indicates that a property possessed by the 
cortex is the combining of a large, though exhaustible, number of move- 
ments, belonging to this and that restricted portion of limb, face, or other 
motile part, into sequences of very great variety, sequences in which 


178 Leyton and Sherrington 


members of the same group of elementary movements follow now in one 
order, now in another, according as the point of cortex stimulated is 
chosen now at one place or now at another not too far apart, and 
influenced also by the stimulations that have been more immediately 
precurrent. It is the isolated and restricted character of the primary 
movements elicited by punctate stimulation of the cortex, or, to repeat 
the term introduced above, their fractional character, which makes so 
equivocal any purpose that an observer, who would interpret their purpose, 
can assign to them. Such a movement as the extension of the index finger 
can serve many purposes, so, again, a closure of the lips, or a retraction 
of the tongue, or flexion of the ankle. Some of the facial movements 
observed suggest mimetic acts, some the acts concerned with feeding; 
some, such as the narrowing of the glottis, might be mimetic on one 
occasion, deglutitional on another. But the combinational sequences are, 
so to say, eloquent of purpose in most instances. The large variety of 
partial, though discrete and in themselves perfect, movements of separate 
portions of the bodily framework, evocable by localised point-to-point 
stimulation of the motor cortex, and the multiform combinations which 
these assume under cortical reaction and the rich mutual associations of the 
cortical motor points which the physiological phenomena of “ facilitation ” 
and “deviation of response” reveal, are suggestive. They lead to the sup- 
position that from movements of locally restricted parts, e.g. movements of 
a finger or of a limb-joint (movements themselves discrete and individually 
separable in the motor cortex), the upbuilding of larger combinations varied 
in character and serviceable for purposes of different and varied kind, 
prehensile, defensive, locomotor, mimetic, masticatory, deglutitional, orienta- 
tional (in von Monakow’s (80) sense), ete., is one of the main offices per- 
formed by the motor cortex. The functional properties of this cortex seem 
specialised for that end. It appears at first sight surprising that a motor 
nervous organ relatively so high as is the cerebral cortex in the nervous 
hierarchy, where the power to deal with large integrated complexes of the 
motor machinery might be expected, exhibits on actual examination a 
representation, still more or less discrete, of relatively small and “partial ” 
movements. And in the motor cortex this discrete “representation” of 
small local items of movement, each highly co-ordinated with others yet 
separably elicitable, instead of becoming less evident with ascent to the 
higher types of hemisphere, becomes more so. Thus, it is more evident in 
cat and dog than in rabbit, more evident in the macaque than in cat or dog, 
in baboon than in macaque, in gibbon than in baboon, and in the chimpanzee, 
orang, and gorilla than in gibbon. It would seem that in order to preserve 
the possibility of being interchangeably compounded in a variety of ways, 
successive or simultaneous, these movements must lie, as more or less dis- 
crete and separable elements, within the grasp of the organ which has the 
varied compounding of them. To draw an analogy merely illustrative, the 
synthesis of the proteins of the body requires that certain metabolic organs 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 179 


must have, lying at their hand, the numerous amino-acid constituents of 
proteins; for that purpose the food proteins, split up into constituent 
chemical sub-groups more or less freed one from another, are presented to 
the synthetic organs for varied re-grouping in the re-synthesis which follows. 
The motor cortex appears to be par excellence a synthetic organ for motor 
acts. How does the motor cortex obtain these fractional and partial move- 
ments on which work its powers of varied synthesis? Simpler co-ordinated 
elements, such as flexion of a single joint, e.g. knee or elbow, can be safely 
assumed to lie ready to its hand in the bulbo-spinal mechanisms. But 
the higher of the compounded movements which those mechanisms give 
tend, if judged from the spinal and decerebrate dog and cat, to be 
compounds exhibiting total flexion or total extension of a whole limb. In 
the limb movements evoked from the anthropoid motor cortex, flexion of 
one joint may go either with flexion or with extension of another. The 
motor cortex may therefore obtain the partial and fractional movements 
it so variously weaves together by, to a certain extent, breaking up com- 
pounds already constructed by lower centres. Such analytic power may 
be a property of its own, or of some other, perhaps subcortical, organ with 
which it keeps close touch. Such synthesis involves time adjustments 
as well as spatial adjustments. The bulbo-spinal axis also, of course, syn- 
thesises motor acts. But the difference between the constructive planes 
of the two is considerable. Bulbo-spinal synthesis constructs in the main 
those locally restricted but co-ordinate movements which the cortical 
synthesis finds ready to hand as elements for it to work with. The bulbo- 
spinal organ taken as a whole does, even in types so high as dog and cat, 
synthesise in addition to the local elementary movements a not inconsider- 
able number of more complex ones, such as respiratory, defensive, and even 
locomotory. But comparison of the synthetic capacity of the bulbo-spinal 
organ with that of the motor cortex reveals a great excess of synthetic 
capacity in the latter, as evidenced by the variety and multiform scope of 
the motor acts and sequences it builds up. Especially is this so when it 
is borne in mind that many acts which, when naturally performed, are 
bilateral, are, when excited by stimulation of one motor cortex, essentially 
unilateral, indicating that the two motor cortices have to be regarded 
as in many respects a single organ when in natural operation. Together 
they form, in such an animal type as the anthropoid ape, an organ for 
synthesis of movements—and of postures—on a vast scale. Phenomena, 
such as “reversal of response,” “ facilitation,’ and “deviation of response,” 
prominent in cortical responses, and accounting for the functional instability 
of cortical motor points, are indicative of the enormous wealth of mutual 
associations existing between the separable motor cortical points, and those 
associations must be a characteristic part of the machinery by which the 
synthetic powers of that cortex is made possible. The motor cortex 
seems to possess, or to be in touch with, the small localised movements as 
separable units, and to supply great numbers of connecting processes 


180 Leyton and Sherrington 


between these, so as to associate them together in extremely varied com- 
binations. The acquirement of skilled movements, though certainly a 
process involving far wider areas (cf. v. Monakow) of the cortex than the 
excitable zone itself, may be presumed to find in the motor cortex an 
organ whose synthetic properties are part of the physiological basis which 
renders that acquirement possible. What has been termed above the 
“functional instability” of cortical motor points seems but one aspect, 
revealed to experiment, of the many-sided motor synthesising which this 
cortex can effect. Such “instability ” may be a means used in those cortical 
readjustments which the experiments of Osborne and Kilvington (35) and 
of Robert Kennedy (23) prove to take place where, after the experimental 
crossing of nerve-trunks, willed movements of normal effect are practically 
restored. As Franz (17) and Bayliss (1) point out, the “instability ” may 
serve as part of the basis on which is founded the educability of the cortex. 


III. EXPERIMENTS BY ABLATION. 


Ablation-Experiment 1. Ablation in Arm Area of Left 
Hemisphere, Chimpanzee (figs. 19, 20). 


Troglodytes niger, f,strong, adult; tame. Not infrequently walks 
erect. Accepts fruit, etc., with either hand, no apparent preference of right. 
After food picks teeth with extended index of either hand. Generally 
walks quadrupedally; a common posture is the semi-erect, the support 
from front-limbs being given by knuckles touching floor. Shakes hands 
with either hand, and occasionally the grip of the hand is then felt to be 
very powerful. Picks up nuts with deftness, but the thumb is less used 
in such movement than might be expected; it seems too short to help the 
fingers for things requiring the finger tips. Sleeps with arm under head 
for pillow, generally with body not on side, but fully supine. When 
wanting to attract notice it has a habit of stamping one foot or both feet 
on the floor. ' 

March 26, forenoon.—Under deep chloroform anesthesia the left 
hemisphere exposed through enlarged trephine hole over lower part of 
centralis region. The lower part of arm area of cortex and the upper part 
of face area explored by unipolar faradisation. The stimulation results 
mapped (fig. 19) and recorded. Then the whole of the area yielding primary 
movements of thumb, index, fingers, wrist, and elbow excised to a depth 
of about 7mm. The excised field included the anterior wall of centralis 
fissure but not the posterior wall, because no motor responses were elicited 
from it or from the free face of post-centralis gyrus. Wound closed 
aseptically. 

Afternoon.—The animal since recovering from the operative narcosis 
has eaten two bananas, and is lively. The right arm, which it seems sur- 
prised to find disability in using, shows marked wrist-drop. It moves right 
elbow and shoulder imperfectly, but with less imperfection at shoulder 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 181 


than at elbow. It moves right fingers little, and thumb and index hardly 
at all. There is a frequent twitching adduction of right thumb. Animal 
seems unable to grasp with right hand; the right hand slips on the 
vertical bars of the cage when animal tries to hold them by it. 

March 27.—The movement of right shoulder is obviously defective. 
The condition of right hand and elbow is the same as yesterday; but the 
animal rarely now attempts to use the hand for grasping, etc. ; it seems to 
have learnt its disability in regard to that hand, and to do without it. 
Animal seems very well otherwise, and is active. Wound looks well. 

March 30.—The movement of right shoulder has improved, but other- 
wise the condition of right arm remains apparently the same as on the day 
after operation. The animal was seen sleeping with its head pillowed on 
right arm, as was not unusual with it before the operation. 


Fic. 19.—Map showing the cortex area ablated in ablation-experiment 1. 
The numerals indicate the responses obtained from it and its neighbour- 
hood. Dotted lines indicate the edges of the ablation, the top one that 
of May 3, the others those of March 26. The numerals refer to the 
‘list of motor responses,” pp. 148-154. 0 denotes that at no time did 
stimulation of the point so marked evoke any response. 

April 26.—Right shoulder seems perfect in all movements, so also 
elbow. Animal often supports itself in the quadrupedal posture by one 
fore-limb, while the other is used for feeding, etc.; for such support the 
right arm is employed seemingly as often as the left, the hand resting on 
the knuckles, and the support involving fixation of elbow in extension and 
of somewhat protracted shoulder. Wrist is moved well, and if any wrist- 
drop is present it is slight, although that wrist is often postured in a some- 
what drooping pose, but questionably more so than usual. The three 
ulnar fingers seem perfectly strong and good in all actions. Index is 
imperfectly moved ; it is generally flexed along with flexion of the ulnar 
three fingers, but without much strength. Nor does it follow the flexion 
of the ulnar three digits perfectly, for occasionally when the vertical bars 
of the cage are grasped by this hand the ulnar three fingers clasp the 
bar, but the index, although incurved, is curved not round the bar, but 
between the bar and the palm. This does not happen with the sound 
left hand. Neither is index of right hand ever seen to be moved in- 
dependently of the other digits, although that is frequently the case in the 


182 Leyton and Sherrington 


left hand. In the left hand index is often both extended and flexed inde- 
pendently of the other fingers, but never in right. Thumb: this digit in 
the normal (left) hand is used less than might have been supposed; as 
mentioned above, the thumb seems too short to be very competent for 
opposition to the other digits, at least fur many purposes. In the paretic 
right hand the thumb slightly but indubitably combines with the other 
digits in a general grasp with the hand, but this grasp movement is in 
reality less abnormal to inspection than it proves to be when the grasp is 
felt. The animal had been taught to shake hands, and with either hand. 
When one shakes hands with its right hand, one feels that it exerts very 
little compression or force with thumb of that hand: whereas, when one 
shakes hands with its left hand, the compressive force of the left thumb is 
felt to be good and considerable. Yet the right thumb is employed by 
it to a far from negligible degree. Thus it frequently employs the thumb 
of right hand in holding a banana, an apple, ete., with that hand; also in 
peeling a banana, the fruit itself being held in the left hand. Occasionally 
when eating fruit it holds the fruit with both hands, the right contributing 
seemingly an equal share of manipulation with the left. Thus, on one 
occasion, the right thumb was clearly seen to be employed with successful 
force to break a banana open, the ends of the fruit being held each with 
one hand, the thumb of each hand pressing down on the convexity of the 
banana from above, and so breaking the fruit. Sometimes the animal 
feeds from fruit held to the mouth by right hand alone. The ulnar three 
fingers of right hand are well used, not only for grasping, but apparently 
for all their full variety of movements. 

May 38.—Condition of right arm appears the same as when last note 
was written. Animal lively and well. The scalp wound has been com- 
pletely healed for more than a fortnight. 

Animal anzsthetised, and the left hemisphere re-exposed in the same 
situation as before. Faradisation of the old lesion yields nothing. Stimula- 
tion of cortex adjoining it in front and behind yields nothing. Faradisa- 
tion of post-centralis for the whole exposed extent of it yields nothing. 
Faradisation of precentralis adjoining the lesion above evokes brisk and 
strong retraction of shoulder, but no movement of wrist or hand, and ques- 
tionably any of elbow. Faradisation of precentralis adjoining lesion below 
evokes retraction of opposite angle of mouth, especially of upper lip. 

The old lesion was re-excised and slightly extended in depth, and the 
cortex adjoining its upper edge was excised for a 3-mm. strip. The wound 
was then closed aseptically. 

May 4.—Animal very well and active; feeds eagerly, dances about and 
stamps on floor to attract notice; climbs about, coos, and calls, etc. Face 
perfectly normal. Right arm, including hand, is used as freely and well 
as before the last operation, but there seems somewhat less free use of the 
shoulder. The limb is, however, used for climbing, swinging, holding food, 
etc., apparently just as before. 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 183 


May 5.—Same condition. 

May 10.—Right shoulder seems to be fully used now; the second 
operation had seemed to impair its movement to some extent. Hand, etce., 
used freely, and seem to have been in nowise impaired by second operation : 
they seem in the same condition as recorded in the note written on 
April 26, certainly no worse. 

May 14.—Animal] well. Wound is practically healed. Animal deeply 
narcotised with chloroform; whole centralis region of left hemisphere 
exposed, faradised point for point, mapped, and results recorded. Results 
of stimulating cortex adjoining borders of lesion same as before. Lesion 
measured 10 mm. along lower horizontal border, 13 mm. along upper 
horizontal border, and has a vertical length of 14 mm. While the animal 
was being put under the anesthetic it was noticed that it clenched its 
right hand vigorously on several occasions, exerting considerable force with 
fingers and thumb. 

Left hemisphere’s centralis region then exposed and explored. Animal 
then killed with chloroform. 

Bulb and cord examined by Marchi method revealed a heavy degenera- 
tion in the left pyramidal tract (fig. 20). In the pyramids the degeneration 
was entirely confined to the left pyramid. At the region of the most 
anterior part of the pyramidal decussation some of the degenerated fibres 
are seen to be among the very first to decussate (fig. 20). In the left 
pyramid the degenerated fibres, although scattered over the whole cross 
area of the pyramid, were somewhat less numerous, in comparison with 
normal fibres, at the ventral lateral angle than elsewhere. In regions of 
the decussation, where the decussation is in full progress and the degenerat- 
ing fibres are undergoing that re-arrangement in large bulk, it can be 
clearly seen that a certain few of them pass slanting dorsally and toward 
the left into the dorsal part of the lateral column of the ipsilateral side 
(34, 41, 25), although the vast majority cross to the lateral column of 
contralateral side. In the upper cervical region (fig. 20) the degeneration 
consists of a heavy crossed pyramidal field, occupying most of the contra- 
lateral lateral column except for a well-marked border zone and for a 
ventral area. In the 2nd and 3rd cervical levels a few degenerated fibres 
lie at the extreme edge of lateral column for a short strip about midway 
between the dorsal and ventral roots. Many degenerated fibres lie in the 
reticular formation at the base of the dorsal grey horn. In the ipsilateral 
half of the cord there exists a slight and scattered degeneration in lateral 
column, occupying about the same area as that of the crossed pyramidal of 
the contralateral side (16, 40, 41, 25). In the ipsilateral ventral column 
there is a well-marked ventral pyramidal tract (19, 42) degeneration 
bordering the whole length of the cross-section of the lip of the ventral 
fissure (figs. 20, 21). 

In lower cervical region the crossed and uncrossed lateral column 
degenerations have become very distinctly less heavy, although their areas 


184 Leyton and Sherrington 


relatively to that of the lateral column remain about the same as higher 
up. They also retain about the same proportions relatively each to the 
other. In the ipsilateral ventral column a “ direct pyramidal tract” 
degeneration is still marked, but its area is smaller, and is confined to the 
deeper part of the side of the ventral fissure. Below the brachial enlarge- 


Fic. 20,—Outlines (x3 nat. size) of cross-sections of the bulb and spinal cord 
of chimpanzee, showing the pyramidal-tract degenerations after lesion in 
the arm area of the left hemisphere, in ablation-experiment 1. 


ment the degenerations are seen to have become much smaller. In the 
mid-thoracie region no degeneration is detectable in the ipsilateral lateral 
column, but in ipsilateral ventral column a few degenerated fibres are still 
obvious near the ventral lip of the fissure. In the contralateral cord-half 
a faint scattered degeneration is still obvious occupying the usual pyra- 
midal-tract area. This latter degeneration is still detectible, although 
much less, at the 13th thoracic level, but lower than that is not recognis- 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 185 


able with certainty. In the lumbar enlargement no degeneration can be 
detected. 

The ventral horn of grey matter in the lower part of the brachial en- 
largement shows, especially in 7th and 8th cervical segments, a marked 
difference between the right and left sides. On the right side, the whole 
of the cross-area of ventral horn has scattered through it many degenerat- 
ing fibres of very minute size; they give a “peppered” appearance to the 
grey matter there, in contrast to the ordinary clean appearance of the 
corresponding grey matter of the left half of the cord. The peppering is 
perhaps most marked in the dorsolateral and ventrolateral cell-group 


R L 


Fic. 21.—Microphotogram of direct pyramidal tract degenerated, along lip of ventral fissure, 
at 3rd cervical segment. Marchi preparation. Ablation-experiment 1. 


regions. It is certainly least in the medio-ventral cell-group. Sections 
stained with Marchi show these degenerated fibres in the grey matter but 
slightly, although, when aware of them, one can detect the presence of a 
number of them by that method, The degeneration in the ventral horn is, 
however, much better revealed by the Schafer (38) combination of the 
Marchi and Kulschitzky methods; the minute blue-black ring surrounding 
the pale axis cylinder, which many of the very small fibres in the grey 
matter give by that method, when seen in cross-section, is altered to a 
minute blob containing no axis cylinder. In other words, the fine col- 
laterals are degenerated, and their sheaths, with that element of it which 
the hematoxylin stain after the mordant tinges deeply, is broken up, and 
the axis cylinder also; and this kind of minute degeneration is scattered 


186 Leyton and Sherrington 


widely and liberally through the ventral horn. Our sections have not 
detected changes in the perikarya of the motoneurons, even where the 
minute nerve-fibre degeneration is most heavy. 


Ablation-Experiment 2. Removal of part of the Arm Area 
from both Hemispheres (fig. 5, A, B, C, and fig. 22). 


Troglodytes niger, $, of Kola-kaamba type; arrived at laboratory 
from W. Africa, March 26, in rather cramped cage, but perfectly healthy. 
Transferred to roomy laboratory quarters, and allowed in and out of his 
new quarters. Habits observed; after eating, picks teeth with nail of 
extended index finger, usually of left hand, but no clear preference in use 
of left hand over right, employing right and left seemingly equally, except 
for doubtful preference of left in picking teeth. Accepts fruit (bananas, 
apples) with either hand; often stores the fruit so taken. Another chim- 
panzee, § , in the same quarters, was at once treated as a friend; on receiv- 
ing biscuit, Paddy, who did not care much for biscuit, gave most of it at 
once to his “ girl friend,” handing it to her. Occasionally he was seen to kiss 
her, perhaps a trick learned on voyage. Not rarely walked fairly erect with- 
out touching floor with fore-limbs at all, especially when free in the room 
outside the cage, but sometimes in the cage. When wanting to attract 
attention, or vexed at not getting his own way, stamps one foot or both 
feet on the floor. Makes a considerable variety of sounds, especially a 
sucking noise, with lips protruded. Sleeps with one arm under head for 
pillow, lying on side. Dungs in one corner of his cage. 

lst Operation, January 3.—Has been in laboratory more than nine 
months. Chloroform and ether mixture (equal parts) administered, and 
taken very well; loud snoring when under deep narcosis. Trephined on left 
side; the hand area of left cerebral hemisphere exposed through the suitably 
enlarged trephine hole. Dura mater turned back in flaps; arachnoid care- 
fully opened, allowing much subarachnoid fluid to escape. 

The exposed convolutions thus freed from fluid were stimulated. Uni- 
polar faradism was used for the most part, with occasional application of 
the ordinary double-point electrodes. Temperature of room 29° C. through- 
out experiment; temperature in rectum 374° C. at commencement and 
37° C. at end of operation. Ether narcosis maintained throughout. 

The cortical region exposed included the precentral and _post-central 
convolutions above and below the inferior genu of central fissure. The 
post-central was carefully explored by faradisation with both forms of 
electrodes, and with the same strength of stimulus as was employed for the 
precentral, but no movement was at any time elicited by the stimulations, 
although the similar stimuli applied to the precentral evoked movements 
regularly. The whole vertical length of precentralis, as far as it was laid 
bare, was found excitable, but the anterior portion of its horizontal width 
did not respond with such certainty as did the rest. The exact topography 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 187 


in it of a number of points with the movements they yielded was carefully 
mapped. This record is reproduced in fig. 5, A. Focal epilepsy was very 
readily provoked; it did not, however, spread; e.g. on several occasions 
when an index finger point was excited the “epilepsy” remained confined 
to index finger. Although faradisation of post-centralis did not provoke 
any movement, it did, when the narcosis was light, seem each time to arouse 
the animal. 

The part indicated by the enclosure within the dotted line on the map 
(fig. 5, A) was then extirpated: care was taken to include the whole anterior 
wall of the sulcus centralis, i.e. down to the bottom of thesulcus. The wound 
was then aseptically closed and surgical dressing applied. On the animal’s 
recovery from the narcosis it was noted that right wrist was “dropped” 
and that the thumb seemed quite paralysed, but that a little flexion and 
extension of the three outer fingers occurred from time to time. The 
shoulder seemed slightly affected, but the elbow not at all. No trace of 
paresis of face or leg. The following morning the animal was found active, 
feeding well, and in good spirits. There was no trace of facial or leg 
paresis. Flexion or extension of the three ulnar fingers of right hand 
occurred, but seemingly only when the hand was placed in contact with 
some object, e.g. bars of cage. Movement of the thumb was seen only 
under circumstances when it might be passive; indubitable active motion 
of it was not seen. Movement of index finger was never seen. The hand 
hung helplessly prone, and dropped at the wrist joint. Power to elevate 
the shoulder seemed defective. No paresis noted at elbow. The defect at 
shoulder probably seemed greater than it really was, for in the afternoon 
on the cage door being set open, the animal ran out, and in climbing up the 
outside of the cage it was seen to raise and protract the right arm well at 
the shoulder. The grasp by the three ulnar fingers appeared to improve 
during the day. In regard to the sensations of the right hand, clear 
evidence was obtained that bending back either the thumb or little finger 
was felt, and was unpleasing to the animal. 

Eight days after the operation. Small pieces of fruit, e.g. bits of a 
grape, were fairly accurately picked up by the right hand. There seemed 
a liability to misplace the whole hand. Fifteen days after the operation 
great improvement had occurred in the use of the limb; a cursory examina- 
tion would hardly detect any defect of movement in it; the wound had 
completely healed. 

2nd Operation, March 3.—Sixty days after the first operation. The 
animal had recovered full use of right hand and arm; the sole remaining 
defect seemed a slight clumsiness of the digits, revealed by testing it in the 
picking up of maize-grains, etc. But during the last previous eight days 
it has seemed that the pose of the right hand when at rest indicates some 
slight over-extension at the metacarpo-phalangeal joints. A second opera- 
tion was now undertaken. The same region of the left hemisphere as in the 
first operation was re-exposed. The dural flap was turned upward toward 


188 Leyton and Sherrington 


mid-line. Main blood-vessels marked in the map of the former operation 
were re-found above and in front of the scar, and were unaltered from their 
previous appearance. Faradisation of the cortex along the lower edge of 
the old lesion evoked no movement in hand, but retraction and raising of 
right angle of mouth, and at one point quite regularly a brisk turning of 
neck and head toward the opposite side. Faradisation by plunging the 
unipolar electrode into the soft sear, even when the penetration amounted 
to 1 em., failed to evoke movement. Precentral gyrus above the lesion was 
explored up to the upper genu; it gave the same results as at the previous 
examination two months before. Post-central gyrus was also re-explored, 
and no movement could be elicited by currents of even greater intensity 
than those sufficing to evoke response from the uninjured parts of pre- 
centralis. The old scar was then entirely cut away, and the old lesion 
deepened everywhere by further ablation ; and the old lesion was increased 
upward by removing part of the gyrus previously uninjured as far as the 
line marked 3. iii. in the map (see fig. 5, A). 

On recovering from the operation narcosis the animal showed no 
distinct aggravation of any paresis there might have been remaining in the 
right arm. The hand was at once used well to grip the bars of its cage. 
The animal often hung suspended by the right hand and arm; and this 
was the better demonstrated, because the animal seemed excited, and was 
for a few hours particularly active. Small objects, e.g. bits of grape and 
such as had been previously used for testing, were picked up well, and to 
all appearance as well and readily as the day before the second operation. 
Sometimes the right arm seemed to be used not quite so freely or well as 
before this second operation, but the difference, if really existent, was a very 
slight one. There was no obvious evidence of astereognosis. There was 
no obvious paresis at elbow or shoulder, and no trace of paresis of face or 
leg. Next day, the animal doing very well and being very active, the 
movements of right arm were thoroughly examined. No difference was 
detected between its existing motility and that obtaining before the last 
operation. Thumb was well and accurately adducted and abducted ; index 
was similarly well flexed and extended; and both were used quite success- 
fully to all appearance. On March 5th, forty-eight hours after the second 
operation, the animal was seen using the tip of the right index finger, 
the finger being isolatedly extended, for picking its teeth. Certainly no 
recrudescence of paresis in the right arm was observable. 

April 1.—The wound has been completely healed for some time. No 
recrudescence of paresis has at any time ensued in result of the operation 
of March 3rd. The tendency to contracture noted shortly before the second 
operation has become more pronounced. The arm tends to be kept 
partially flexed at elbow, and there is some postural over-extension at 
metacarpo-phalangeal joints, with some flexion at the phalangeal joints of 
all the digits except thumb. 

3rd Operation, April 2.—The left hemisphere had been the field for 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 189 


the previous two experiments. The right was now exposed at the level of 
inferior genu of sulcus centralis. The field of cortex exposed embraced 
about a third of the length of both precentral and post-central gyri, and 
the exposed portions of these were carefully faradised from point to point. 
From post-centralis the faradisation never evoked any responsive move- 
ment, despite repeated trials, often with very strong currents applied both 
with the unipolar and with the bipolar electrodes. Precentralis, on the 
contrary, was an “excitable” field continuously throughout the whole 
portion of its length exposed to view, i.e. from 5 mm. below superior genu 
to about 6 mm. below inferior genu. The topography of a number of the 
points yielding the movements and the movements so obtained were re- 
corded and mapped (fig. 5, B). It is noted that in proceeding with the 
application of stimuli to a series of points taken in successive order from 
above downward, i.e. passing away from mid-line responsive, movement of 
elbow could be evoked as a “leading” (primary) movement quite down to 
genu inferius; whereas when the successive stimuli to the points pro- 
ceeded in the opposite direction (upward), movement of index and thumb 
was evoked as the “leading” (primary) movement, even when the stimulus 
reached points up in the shoulder area of the cortex near genu superius. 
From a narrow area just below genu inferius there was elicited closure 
of the lids of the opposite eye unaccompanied by any other movement. 
Careful search was made for evidence of movement in the right arm on 
stimulation of this the cortical area for the left arm, in order to test the 
supposition that the recovery of the right arm movements might be ex- 
plicable by supplementary functions for right arm taken over by the 
cortical field for left arm. Even with very strong and diffuse (widespread 
bipolar electrodes) stimulation, let alone moderate and weak with the uni- 
polar electrode, never was any trace of movement of right arm evoked by 
excitation of this the motor arm area of the right hemisphere. The move- 
ments elicited in left arm were, however, very various and vigorous. 
Finally, the whole of the area which under faradisation had provoked 
“leading” (primary) movement in fingers, thumb, wrist, and elbow was 
then extirpated by the knife to a depth of about 8 mm., and the floor of 
the ablated area cauterised superficially with the electro-cautery. The 
wound was then closed with full aseptic procedure (fig. 5, B, limit lines 
of lesion shown). 

‘On recovering from the operation narcosis the animal became some- 
what excited and very active in its movements. Not the slightest re- 
crudescence of symptoms of paresis and clumsiness in the right arm was 
detected. In the left arm the opportunity for seeing impairment of move- 
ment was excellent, because the animal (and the same condition was re- 
peatedly noticed in other experiments on cortical ablation) was evidently 
slow to realise that he could not use the limb as before. The animal spent 
nearly five minutes in trying to pick up individual grape-berries placed on 
the floor of his cage. There was evident inability to move thumb and 


190 Leyton and Sherrington 


index of left hand, and very little ability, although some, to move the 
three ulnar digits. The animal seemed imperfectly aware of where his 
thumb and index actually might be. Yet in climbing about his cage he 
hooked on to the bars with the ulnar fingers of left hand curved to a 
rather open claw. The wrist was clearly “dropped”; at elbow and at 
shoulder there was little obvious impairment. Next day there was distinct 
improvement in the motility of left hand and wrist. There was less wrist- 
drop, and the index finger was seen to be moved at times. The animal was 
in good general condition, and climbed actively about the cage. It was 
therefore determined to ablate more of the arm area of right hemisphere. 

4th Operation, April 3.—The animal was again put under deep 
narcosis, and the central gyri of the right hemisphere were exposed for 
the middle portion of their length. The cortex of precentralis was found 
excitable down to the very margin of the area extirpated the day before. 
From a point in the arm area close above the extirpation, field movement 
of little finger was obtained by strong faradisation; this was a primary 
movement of the little finger unaccompanied by movement elsewhere. 
From arm area close above the extirpation, field movement of other fingers, 
and slightly of thumb and index, was also obtained, but always as a 
secondary movement, never as a leading or primary one. “Epilepsy” of 
arm when provoked sometimes spread to the ulnar fingers, but never to 
thumb or index. The area of yesterday’s extirpation was itself tested by 
thrusting the electrode 10 mm. into it; the electrode, though buried in clot, 
must have reached the floor of the ablation hole, and movements of thumb 
and index were then obtained, but were very weak and uncertain. Out- 
side the lesion area the precentralis was found, as on the previous day, to 
present a continuous excitable field. From the post-centralis, as yesterday, 
no responsive movement was obtainable. A map was made, fixing the 
topography of a number of the excited points in precentralis. The area of 
cortex indicated on the figure as bounded by fissura centralis and the dotted 
line above (fig. 5, B, limit line marked 3, iv.,) was then excised to the same 
depths as yesterday. After this ablation the cortex of precentralis above 
the freshly ablated piece was tested with faradisation and found to be still 
excitable down to its very edge, but from it shoulder movements only were 
elicitable, and there occurred no “march” or spread of the movement to 
other portions of the limb. 

On recovering from the operation narcosis the animal exhibited marked 
wrist-drop, more so than yesterday; but active movements of the wrist 
were noticed to occur from time to time. The ulnar fingers were not so 
successfully moved as before the operation, though they were still slightly 
moved at times. A little movement was still observed occasionally in 
thumb and index. Using the ulnar edge of hand with its partially 
flexed ulnar fingers as a sort of claw, the animal conveyed a grape-berry 
from the hand offering it to its own lips. Its lips took it with difficulty, 
because the pronated hand came with its radial edge toward the mouth, 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 191 


the berry lying at the far end of a short cone formed by the claw-like 
hand tapering to its ulnar edge. Elbow seemed slightly defective in motor 
power. Distinct, though slight, impairment of motor power was noted at 
shoulder. There was not any trace of recrudescence of paresis of the 
right arm. 

Next morning considerable improvement was obvious in the motility 
of left arm. Elbow and shoulder were no longer noticeably affected. 
The arm was used fairly well for swinging from the roof of the cage. It 
was clear that the ulnar fingers were used fairly well in willed movements. 
Wrist-drop was severe, and seemed quite as marked as yesterday afternoon. 
Two days later the left arm had improved further. Thumb and index 
were evidently better and more freely employed. The animal when induced 
to put forward its left arm for fruit, etc., offered to it does so by thrusting 
the arm fairly skilfully, though with seeming hesitation and some effort, 
between the cage bars. The paretic wrist is prone. The grape-berry is 
taken by the three ulnar fingers, more flexed than formerly, assisted now 
by the index; the thumb also was often moved in the manceuvre, but did 
not practically assist. 

April 7.—Further improvement in motility of left arm. Since the 
right cortex operation, which impaired the motility of left arm, the motility 
of right arm has notably increased, and right arm has been much more 
frequently employed than before. No paresis remains detectable in it; 
and hand, and the whole arm, are now repeatedly employed for all their 
usual purposes. This morning, after its breakfast, the animal sat and 
picked its teeth with the isolatedly extended index finger of right hand. 
It was seen also to pick and scoop out the furrows of the pinna of the 
left ear with right index finger. When making an effort to take with the 
left hand a small object, e.g. maize-grain, there occurs frequently an 
accompanying strong contraction (flexion) of the fingers of right hand. 
The converse has not been noticed to occur. 

April 8.—Further improvement in motility of left arm, less wrist-drop, 
better movement of index and thumb. 

5th Operation, April 8.—Under deep narcosis the arm area and the 
circumjacent cortex of left hemisphere was laid bare. Gyrus post-centralis 
was then tested by faradisation to see if, especially at the levels opposite 
the excised portion of arm area, it had acquired motor responsiveness to 
the electric stimuli, but no motor responses could be elicited from it. From 
precentralis above the lesion, from the edge of the lesion right up to the 
trunk area, between arm area and leg area, repeated excitation elicited, and 
elicited easily, movements of shoulder, but of no other part of arm. Move- 
ments thus evoked in shoulder were never on any occasion accompanied by 
or followed by movements of elbow, wrist, fingers, or thumb. “Epilepsy ” 
was frequently obtained, but remained confined to shoulder, never spreading 
to other parts of the limb. A map was made recording the topography of 
a number of points at which carefully observed movements were evoked. 

VOL. XI., NO, 2.—1917. 13 


192 Leyton and Sherrington 


The precentralis throughout the region explored outside the lesion with 
the electrodes was found to consist of a continuous field of excitability for 
the whole length examined in it. The strip of cortex above the former 
lesion was then excised to the limit shown in fig. 5, A, by the broken line 
marked 8, iv. From the old lesion the electrodes never obtained responses, 
although plunged deeply into the tissue, and although both the single and 
double electrodes with strong stimuli were used. 

On recovery from the operation narcosis the animal showed no impair- — 
ment in the motility of right arm. Grapes and pieces of grape and maize- 
grains were picked up as successfully as before. Once it was noted that 
the animal having picked up a grape-berry carried it by right hand to the 
base of the forehead at root of nose, seemingly instead of to the mouth. 
But on many other occasions the morsel was carried by the hand accurately 
and quickly to the mouth. The following morning, April 10th, the animal 
was seen to pick something out of the inner canthus of the left eye with 
the tip of the right index finger. The left arm had a good grasp; there 
was much less wrist-drop; elbow was certainly moved freely; but there 
seemed some misjudgment of the position of the left hand. 

May 4.—The animal now uses both hands and arms well. Employs 
either hand in feeding himself with banana or grapes. Peels banana, hold- 
ing it in one hand and stripping off the peel with the other. 

Under deep narcosis the central region of left hemisphere laid bare 
(fig. 5,C). Faradisation of centralis anterior above the excised area from 
border of the old lesion to superior genu evoked merest trace of shoulder 
movement and no trace of movement elsewhere in arm, but gave near edge 
of lesion and for some distance above it movement of chest muscles, uni- 
lateral and crossed, and not otherwise interfering with respiratory rhythm 
of chest. Higher still centralis anterior gave movements of abdominal wall, 
also unilateral and crossed. Higher still the leg area gave usual results. 
The cortex adjoining lower edge of lesion evoked retraction of opposite 
angle of mouth with some inversion of upper lip. From the cortex in 
front of the lesion no response was obtainable. From the centralis posterior 
likewise no responsive movement was elicitable when faradised under 
ordinary conditions, but on faradising it opposite the leg area immediately 
after a precurrent stimulation of centralis anterior closely on the same 
level there seemed distinct re-induction of the same movement which had 
just previously been given by the centralis anterior. 

The right hemisphere in centralis region was then similarly examined, 
and gave similar results (fig. 5, B). The animal was then killed. 

Examined microscopically, the bulb and spinal cord revealed by the 
Marchi method the following degenerations (fig. 22):—The pyramids 
exhibited large numbers of degenerated fibres distributed fairly symmetri- 
cally right and left, but distinctly rather less numerous in the right pyramid 
than in the left. In both pyramids the degenerated fibres were rather less 
densely scattered in the extreme ventrolateral portion of the cross-section. 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 193 


Some of the degenerated fibres were among those that entered the pyramidal 
decussation in the headmost portion of the decussation. In the spinal cord, 
at a level midway between Ist and 2nd cervical roots,a heavy degeneration 
exists in the pyramidal region of both lateral columns, but is heavier in the 
right than in the left. In both the degeneration extends out to the extreme 
edge of the lateral column for the greater part of the periphery of the 
column, but is absent from it at the dorsal angle of the column, where it 


Fic. 22.—Outlines ( x 3 nat. size) of cross-sections of the bulb and spinal cord of 
chimpanzee, showing the pyramidal-tract degenerations following lesion in the 
arm area of both hemispheres in ablation-experiment 2. 


scarcely at all invades an area of large nerve-fibres (dorsal cerebellar tract). 
The main degeneration lying in the deeper part of the lateral column joins 
the zonal part of the degenerated area by a narrowish isthmus. A small 
degeneration exists in each ventral column alongside the deeper half of the 
lip of the ventral fissure. A segment lower the configuration of the 
degenerations in cross-section has altered, in that a zonal area in the dorsal 
half of each lateral column exists free from degenerated fibres. By the 
level of the 7th cervical segment the amount of degeneration has obviously 
lessened ; it is clearly less heavy in the left half of cord than in the right. 
In each lateral column it reaches the periphery over a limited small stretch 
about the junction of the ventral third with the dorsal two-thirds of the 


194 Leyton and Sherrington 


column’s edge. A few degenerated fibres lie in both ventral columns 
opposite the deepest part of the ventral fissure. At a level between 2nd 
and 3rd thoracic segments the degenerations in both lateral columns are 
found to be very greatly lessened, especially in the left; and the disparity 
between the right and left lateral column degenerations is more obvious 
than higher up. A trace of degeneration is still detectable in the left 
ventral column at the deep part of the ventral fissure’s lip. At the mid- 
thoracic region there is still some degeneration in the pyramidal region of 
both lateral columns, distinctly heavier on right side than left. In the 
upper lumbar region degenerated fibres are detectable in the right lateral 
column’s pyramidal region, but none are obvious in the left. Below the 
3rd lumbar level no trace of degeneration was discovered. 


Ablation-Experiment 3. Ablation of part of Leg Area 
from Left Hemisphere (figs. 2, A, and 28). 


Chimpanzee, § , rather small, well nourished. 

June 25.—The upper part of gyrus centralis anterior exposed under 
deep anesthesia. After reflecting the dura mater the surface of the ex- 
posed portion of centralis anterior and of the adjoining centralis posterior 
and of the cortex in front of sulcus precentralis superior was explored with 
the electrode up to the near neighbourhood of superior longitudinal fissure. 
Fig. 2, A, indicates the positions of a number of the cortical points tested and 
their responses. No responses were obtained from centralis posterior even 
with the second coil of inductorium pushed up to5cm. A large part of the 
leg area cortex was then excised to a depth of about 8 mm. The excision 
was carried down to the region yielding abdominal wall movements. The 
points marked 262 and 281 lay in the exposed area, but were not excised. 
The points marked 313, 339, 269, 292, and 293 were not exposed at this 
operation. The whole operation was carried out aseptically, and the dural 
flaps reclosed and stitched, and the wound closed. 

On recovering from the anesthesia the animal was active, and showed 
clear paresis in right hind-limb, but it used the limb in climbing and 
moving about the cage. No paresis detected in fore-limb. The impair- 
ment of right hind-limb was evidenced in clumsiness as well as weakness, 
and affected the digits as well as the hip. 

June 26—Animal lively. Seems more aware of defective control of 
right leg than yesterday, when it seemed to trust to it, and be unaware 
of disability in it until it put it to use. Right knee-jerk is distinctly more 
ample and brisk than left. 

June 27.—Animal lively. Evidently more power and less clumsiness 
in right leg. 

June 30.—Animal lively; scalp has nearly healed. Climbs about 
freely, and right leg is much used, and has improved. 

July 10.—Very little noticeable disturbance in right leg in its use as 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 195 


animal climbs about cage and runs about the room. Right knee-jerk is 
the more brisk. 

July 20.— Right knee-jerk exaggerated as compared with left, but 
as the animal's free movements are examined when playing with other 
animals and in going about cage and room, there is no clear evidence to 
inspection of abnormality in the leg movements. The bar is grasped by 


A ) 
ee 


Fic. 23.—Outlines (x3 nat. size) of cross-sections of the spinal cord, 
showing the degenerations following ablation of part of the leg 
area of left hemisphere ; chimpanzee, ablation-experiment 3, 


right foot deftly, and apparently with power, certainly with enough power 
to swing the animal from it. 

July 24.— Animal anesthetised and the central gyri of both hemi- 
spheres examined systematically by unipolar faradisation. The lesion in 
leg area, left hemisphere, includes less of that area than was expected ; 
points 262 and 281, both of them, now yield movement 343. The contrac- 
tion of the lesion which has occurred seems to have narrowed the lesion 
laterally and to have displaced it toward mesial edge of hemisphere, so 


196 Leyton and Sherrington 


that the top border of lesion is a little over on mesial surface, which it 
certainly was not when made. The rest of leg area was explored, and gave 
results a number of which are in the map (fig. 2, A). Centralis posterior 
still remained quite unresponsive to stimulation. Animal then killed with 
chloroform. 

Examination of the bulb and cord by Marchi method showed (fig. 23) 
a degeneration scattered all over the cross-section of the left pyramid, but, 
entirely confined to that. The amount of degeneration appeared to be 
less than that in the other specimen of leg area lesion and than in the 
arm area lesions. In the upper cervical region the contralateral degenera- 
tion at 5rd cervical level shows only a small extension to the margin of 
the lateral column. There is a small scattered degeneration in ipsilateral 
lateral column, and a smaller degeneration still in ipsilateral ventral column 
by the side of the ventral fissure. At the level of the lower part of Ist 
thoracic segment these degenerations, though they have shifted in position 
somewhat, the crossed pyramidal especially having spread ventrally, 
exhibit no obvious diminution. In the mid-thoracic region the ipsilateral 
lateral column degeneration has the appearance of being a little heavier 
than in cervical region, but is quite slight, and any increase in it is dubit- 
able. At the 12th thoracic segment, last thoracic segment but one, the 
ipsilateral ventral pyramidal degeneration has almost disappeared, but the 
crossed pyramidal seems still as heavy as anywhere above, and has 
encroached toward margin of column laterally, and extends farther 
ventrally. At the Ist sacral level the crossed pyramidal degeneration has 
become obviously less, but is still extensive; it has the comma shape noted 
in the other leg area lesion; there is still an obvious ipsilateral degenera- 
tion in the lateral column, but it is quite slight. 


Ablation-Experiment 4. Ablation of part of Leg Area 
of Left Hemisphere (figs. 24 and 25). 


Troglodytes niger, 7, adolescent. 

May 13.—The upper part of centralis anterior of left hemisphere 
exposed under deep chloroformisation, and with aseptic precautions. 
Precentralis gyrus and post-centralis explored by faradisation from the 
superior genu of centralis sulcus up to the mid-line. The motor responses 
noted and their cortical points mapped (fig. 24), they showed the same 
general results as obtained in previous experiments. The leg region came 
down to level of superior genu, which was not well marked, but in its 
neighbourhood the primary movements of the responses were practically 
always of hip. Below the hip area, and meeting it, was a transverse strip 
of the convolution, which evoked movements of abdominal wall. A large 
vein crossed the gyrus horizontally about 2 mm. above the lower edge of 
the abdominal wall area. In the upward direction the hip area merged 
into an area yielding knee movements, and this latter in turn into an 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 197 


area yielding ankle movement and toes movement. Gyrus centralis 
posterior was faradised at points over the whole of its exposed surface, and 
yielded no movements. The leg area was bounded in front by an ascend- 
ing limb of precentralis superior fissure, and above that by a small vertical 
fissure lying farther forward. Large veins entering the superior longi- 
tudinal sinus near the mesial border of hemisphere precluded satisfactory 
examination of that actual border. An incision parallel with the mesial 
border, and only a few millimetres from it, was made to a depth of about 8 mm. 
along the whole antero-posterior extent of the leg area; and another along 
precentralis fissure, and from it to the anterior end of the mesial incision ; 
and another along the centralis sulcus, meeting posterior end of the mesial 
incision ; and finally, a transverse incision across centralis anterior close 
above the large vein at the genu superius. Beginning from the mesial 
incision, the cortex of the whole area enclosed by these incisions was then 


Fic. 24.—Map showing the limits of an ablation in the leg area of 
the left hemisphere of a chimpanzee, ablation-experiment 4. The 
numerals refer to the motor responses obtained; vide ‘‘list of 
motor responses,” pp. 148 seq., supra. The double ruled line 
indicates the position of the mesial border of the hemisphere. 


removed to a depth of about a centimetre, so as to include buried portions of 
leg area cortex in the side fissures (fig. 24). A transverse strip of cortex next 
below this lesion, which had previous to the ablation given hip movements, 
evoked after the ablation no movements of hip or leg, but gave instead 
brisk movements of the abdominal wall, as did the strip of cortex immedi- 
ately below it, which had yielded them prior to the ablation. Centralis 
posterior after the ablation was irresponsive, as it had been before. 

The dural flaps were brought together and stitched; the skin was 
closed, and the whole operation completed, as it had been prosecuted, under 
full asepsis. 

On recovering from the operative narcosis the animal showed slight, but 
distinct, paresis of the right leg; the leg could not support the animal on 
the vertical bars of the cage so well as did the left leg. On landing on 
floor from descending the bars of the cage the right leg did not seem able 
for a moment to support the weight of the body, but yielded under it, so 
that animal lurched to that side, and once rolled over. The weakness 
appeared chiefly in the right ankle. There was apparently clumsiness in 


198 Leyton and Sherrington 


catching hold of the bars with it, not so much due to the digits as to the 
ankle. The right foot, as the animal climbed about, sometimes missed the 
bars. It never grasped the bar so fully as did left. The limb was, how- 
ever, freely moved and used. Next day the animal was lively, and fed 
well; the clumsiness and want of strength in climbing with right leg was 
distinctly less than on the day previous. 

May 24.—The right leg is now well used, although obviously some- 
what clumsy. Wound has healed almost. Animal has no difficulty in 
supporting itself with right leg. 

May 30.—Very little obvious impairment of movements of right leg. 

June 3.—Left hemisphere exposed in centralis region under deep 
anesthesia. The whole of free faces of precentral and post-central gyri 
explored with faradisation. The cortex of leg area below the lesion 
yielded no hip or leg movement, but only movements of abdominal wall 
and trunk, 

The extreme mesial edge and the whole mesial face of the leg area 
were then explored and the motor responses mapped (fig. 24). Animal 
then killed with chloroform. 

Examination of the bulb and spinal cord by the Marchi method revealed 
(fig. 25) a heavy degeneration scattered throughout the left pyramid, but 
confined to that. At Ist cervical segment there is a heavy scattered 
degeneration in the right lateral column occupying the pyramidal tract 
area as usually figured, but also occupying a considerable length of dorsal 
half of the margin of the column; this marginal degeneration is separated 
from the deeper area of degeneration by a dorso-ventral strip containing 
large sound fibres (cerebellar tract), but the marginal and deep areas conjoin, 
especially ventrally, by thinly scattered intervening degeneration. In the 
other (ipsilateral lateral) column a small amount of diffuse degeneration 
extends through the pyramidal tract area. In the ipsilateral ventral 
column, beside the whole length of the lip of the ventral fissure, a ventral 
direct pyramidal degeneration exists. A couple of segments lower the 
degeneration in contralateral lateral column has assumed a different shape, 
the marginal degeneration lying farther ventral, and joined by a narrow 
isthmus dorsally to the main deep-lying degenerate area. Ipsilateral 
uncrossed pyramidal tract degeneration in lateral column is still obvious, 
so also the uncrossed ventral, but latter lies deeper down the lip of ventral 
fissure. In the lower cervical region the contralateral degeneration has 
assumed an oval form, with a long ventral extension reaching ventral 
margin of lateral column. In the ipsilateral side, lateral and ventral 
columns exhibit the lateral and ventral uncrossed pyramidal degenera- 
tions as before. In upper thoracic and in mid-thoracic levels the contra- 
lateral degeneration is marked by the considerable extent of its ventral 
sweep, and the ipsilateral by the gradual shift of the ventral column 
portion of it so as to lie in the ventral portion of the ventral fissure’s lip 
once more. There has been no marked, or indeed obvious, decrease in 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 199 


the amount of the degenerations, either contralateral or ipsilateral, in their 
descent along the cord so far. On reaching the lumbar region the contra- 
lateral degeneration shifts out laterally so as to leave the deepest region 
of the white column next to lateral horn, and to oceupy about the dorsal 
three-fourths of the margin of lateral column. By the level of interval 
between 4th (last) lumbar and Ist sacral segments a great diminution in 
the extent of the degeneration is obvious; the contralateral has assumed a 


~ 1-2th 


Fic, 25,—Outlines ( x 3 nat. size) of cross-sections of the spinal cord of chimpanzee, 
showing the degenerations following on ablation of part of the leg area from the 
left hemisphere ; ablation-experiment 4. 


comma-shaped form (fig. 25), the tail of the comma lying along the margin 
of the lateral column. The deeper part of the degeneration is in contact 
with grey matter only along the dorsal horn. Of the ipsilateral degenera- 
tions, the one in ventral column is no longer present, and that in lateral 
column lies in a situation corresponding with that of the head of the comma 
of degeneration on contralateral side. 

In the lumbo-sacral enlargement, especially in the 4th lumbar and Ist 
sacral segments, the ventral horn of grey matter shows on the right side a 
fine-fibre degeneration similar to that mentioned under the unilateral arm 
area lesion. 


200 Leyton and Sherrington 


Ablation-Experiment 5. Removal of Area yielding Closure 
of Eyelids from both Hemispheres (fig. 12). 


December 2.—Gorilla savagei, young, ¢. Under deep chloro- 
form narcosis trephined over the left hemisphere, lower centralis region. 
With strict aseptic precautions the centralis from superior genu downward 
and the frontal region anterior to it exposed and explored. Closure of 
eyelids extremely readily and regularly elicited as a primary movement 
from a small area which was made up of a seemingly continuous field of 
points, each of which evoked closure of eyelids, especially of contralateral 
eye, usually accompanied or followed immediately by some other secondary 
movement. In this area on some occasions, from a small part of it the 
eyelids-closure was not primary, but followed upon a briefly precedent 
movement of mouth (retraction of angle of mouth contralateral to stimulus). 
The map (fig. 12, A) illustrates with simplification of details the condition 
and area found. The area’s general position in the functional topography 
of centralis anterior was determined carefully by test stimulations of points 
above and below it. The centralis anterior was found to yield a seemingly 
continuous field of motor points from genu superius above to the very tip 
of the centralis fissure below. Characteristic localising points, as found in 
it, are entered on the map (fig. 12, A). Centralis posterior was nowhere 
found excitable. 

Forward of the centralis anterior, in the region exposed, two fields were 
found, giving eyelid movements in addition to the area yielding closure of 
lids in the precentral gyrus itself. Both these frontal fields yielded open- 
ing of eyes, and were functionally characterised by other distinctions, also 
from the eyelid region in precentralis. These distinctions were that (1) 
the responsive movement could be evoked only by stronger faradisation 
than that sufficing to evoke motor response, including eyelid-closure re- 
sponse from precentralis; e.g. precentralis, eye-closure at 12:5 em. of 
second coil; eye-opening from frontal regions at 10°5 cm. of second coil; 
(2) neither of the frontal fields yielding eye-opening offered a seemingly 
continuous field of excitable points, but consisted of scattered points which 
were excitable, yet even these were not excitable so regularly as were the 
eye-closure points in the precentralis; (3) the eye-opening movement was 
usually to all appearance fully symmetrically bilateral, sometimes it seemed 
slightly quicker or stronger contralateral to stimulation, but on the whole 
its bilateral equality was in marked contrast to the decided asymmetry of 
the eyelid-closure movement evoked for precentralis; this latter was never 
observed to be fully equal in both eyes, there being always a detectible 
preponderance of the movement on the contralateral side. Not rarely the 
eyelid-closure from precentralis was confined to the contralateral eye, and, 
when weak, was observed on one occasion to be confined to the contralateral 
eyes lower lid only, and on more than one occasion seemed to be confined 
to the upper lid of crossed eye only. 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 201 


The movement of opening of eye responsive to faradisation of frontal 
cortex anterior to precentralis occurred, as mentioned above, in relation to 
two seemingly separable fields. Of these the lower one (fig. 12, A) lay with 
suleus precentralis inferior not far behind it. From this region the scattered 
points evoked eye-opening, often quickly followed by lateral deviation of 
the pupils towards the contralateral side. The other and higher field lay 
with the superior spur of suleus precentralis medius behind it, and the 
inferior spur of suleus precentralis superior farther behind it still. In this 
upper field the scattered points yielded eye-opening seemingly secondarily 
to turning of the eyeballs to contralateral side. 

After the exposed region of cortex had been carefully explored and 
these results noted and mapped, the whole of the small field yielding eye- 
closure was carefully excised, the limits of the excision being shown in the 
map (fig. 12, A) by a dotted line. 

The dura was then replaced, stitched, and the whole wound closed. 
On recovering from the operative narcosis the animal revealed no indica- 
tion whatever, so far as could be detected, of this lesion in regard to the 
eyes or eyelids. There was, however, distinct, though slight, drawing of 
the mouth toward the left side, and a slight flattening of the nasal fold on 
the right side, i.e. some slight paresis of lower part of face on right side. 
These symptoms were obvious the same evening and the next morning, 
and no others were observable. Two days later the slight facial paresis 
was no longer observable, and no abnormality of eyelids had been noted 
at any time. 

December 6.—Under deep chloroform narcosis trephined over the 
lower centralis region of right hemisphere. Centralis fissure exposed from 
genu superius downward, also the frontal region anterior to it for some 
distance. Precentralis carefully explored with unipolar faradisation, with 
especial reference to eyelid-closure area.. A small continuous field of excit- 
able points giving this movement as its primary response was made out, as 
indicated on map (fig. 12, B). Precentralis above and below also offered a 
seemingly continuous field of motor points; characteristic landmark points 
in the field as noted among others at the time are inserted in the map. It 
was noted that the eye-closure obtained from precentralis seemed rather 
more markedly contralateral than usual, i.e. that the associated closure of 
right eye accompanying closure of left seemed rather weaker than usual. 
As with the left hemisphere explored five days previously, so here with the 
right, a wide field yielding eye-opening was met with, and this field seemed 
separable into a lower and an upper, as in the left hemisphere, and the 
characters of its reaction resembled those already mentioned for the corre- 
sponding area of left hemisphere. It was noted, however, that from two 
points in the lower field the movement of eye-opening was followed by 
distinct convergence of the eyeballs, the convergence being directed toward 
a point not far aside from a plane continuous with mid-sagittal plane 
of head. 


202 Leyton and Sherrington 


The eyelid-closure area was then carefully ablated within the delimita- 
tions marked in the map (fig. 12, B). The dura was replaced, stitched, and 
the whole wound closed with aseptic precautions. On recovering from the 
operative narcosis the animal was noticed not to close the eyelids fully 
when blinking. Blinking was as frequent apparently as usual, and could 
be elicited by suddenly approaching the hand to the animal’s face. 
Stronger eyelid-closure was elicited by touching the eyelids or conjunctiva, 
but even then the eye-closure was not tight, and seemed distinctly less 
vigorous than normal. The eyelid-closures which the animal now showed 
were clearly much less tight and vigorous than had been the closures 
evoked by many of the stimulations applied to the cortex. In these latter 
the skin of the eyelids themselves often was actually wrinkled by the 
closure, but this was never the case now with the closures produced by 
the animal itself, even in response to touching the conjunctiva. 

The next day the condition remained the same as regards eyes. Some 
paresis of the lips was obvious. 

December 8.—‘“The closing of the eyes seems now better; paresis in 
face quite as marked as yesterday ; wound puffy.” 

It was then determined to open up the cortex; this was done under 
deep anzsthesia. The whole of the central region of both hemispheres 
was then systematically examined by unipolar faradisation. Nowhere in 
either precentralis were any further points found yielding eye-closure. 
The eye-closure area of both sides appeared to have been completely 
extirpated. The “motor” area otherwise gave under the analysis results 
in harmony with the results obtained on the two previous gorillas. The 
results were mapped and recorded, but as the observations on the other 
two animals have been given in some detail, and these on this animal 
presented no obvious departure from those, the details are not reproduced, 
although incorporated in the general list of movements recorded in the 
anthropoids examined by us. 


Ablation-Experiment 6. Removal of major part of left 
Gyrus centralis anterior (fig. 26). 

Troglodytes niger, #, very young, weight 2°45 kilog., well nourished ;. 
takes milk, sucking it from bottle. 

July 27.—Left hemisphere exposed under deep anesthesia; full aseptic 
procedure ; the centralis region laid bare for its whole length; gyri centrales 
anterior and posterior explored by faradisation ; no responses evoked from 
post-centralis anywhere, but precentralis offered a seemingly continuous 
field of excitable points throughout its whole length. The mesial face of 
hemisphere was not explored. Protrusion of anus was evoked from 
extreme top of centralis anterior well in front of upper end of central 
fissure. Coming down the length of gyrus precentralis the general results 
were like those obtained in other specimens. The abdomen-chest area 
between leg field and arm field was well evident, there being no spur 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 203 


fissure across the precentralis as there usually is at that part, and the 
great veins over-running the gyrus there being not so large as usual. The 
individual movements of index and thumb were easily elicitable. A narrow 
zone of representation of neck was demonstrable between hand and upper- 
face region at the genu inferius. The various movements elicited from 
precentralis were in no case at all choreiform. “Epilepsy” was evoked by 
strong and prolonged stimuli, but it remained almost confined to the part 
implicated in the primary movement belonging to the focus excited, and 
did not spread. 

The precentralis was then ablated for its whole length, except that a 
narrow transverse strip at its top end was left next to the longitudinal 
sinus; and for its whole width, except that a strip 1-2 mm. wide was left 
along the posterior border next post-centralis, and that the arm annectant 
to middle frontal gyrus and leg annectant to superior frontal gyrus were 
not far trespassed upon. 

The wound was then closed aseptically. 

On recovering from the operative narcosis the animal was seen to be 
hemiplegic in right face, arm, and leg. Next day, 28th, the face was 
asymmetrically posed; the cheek bagged on the right side; the nasal fold 
was less marked right than left; the right half of the mouth was not fully 
closed, whereas the left was. When the eyes closed the play of the upper 
lids seemed bilaterally equal, but the play of the lower edges of the 
palpebral fissures seemed greater and stronger for right than left eye. The 
tongue lay in the mouth, with its mid-line, especially just behind the tip, 
slightly to left of the mid-line of the mouth. The play of the lips, both in 
protrusion and in retraction, was very much greater on the left side than 
the right; indeed, the right half of both lips seemed in flaccid paralysis. 
Careful examination failed, however, to detect any difference in posture or 
movement between the forehead, eyebrows, or upper eyelids, right and left. 

Right hand and right foot were often moved fairly extensively, but 
both were markedly paretic. It was several times noticed that flexion of 
the right fingers and hand accompanied willed movements of the left arm, 
and that folding of the right toes accompanied willed movements of the 
left leg. Movements of right shoulder were poor and weak. On scratching 
the sole of the right foot the digits were extended and spread, but scratch- 
ing similarly the left sole induced no movement. Tickling the right side 
of the face with a straw evoked lively action of the face; similar tickling 
of the left side of face evoked no facial action. 

The pupils were equal; no disturbance of eyeball movements was 
detected, though frequently looked for. 

July 29 (fig. 26).—Animal climbs about cage a little, but practically 
makes no use of right arm, though some use of right leg, holding bars of 
cage fairly well with right foot: is not feeding well. 

July 30.—Refuses food. Right hemisphere exposed under deep narcosis, 
and then explored by faradisation and mapped. Results resemble those 


204 Leyton and Sherrington 


previously noted for right hemisphere in all general features. Left hemi- 
sphere re-exposed ; motor responses obtained at some points from posterior 


Fic. 26.—Very young chimpanzee ; large lesion in gyrus centralis anterior of left hemisphere. 
y young i > larg) gy 


strip of centralis anterior not excised; no motor responses from centralis 
posterior anywhere, thus agreeing with previous examination. Animal 
killed by chloroform. 


Ablation-Experiment 7. Removal of part of Inferior Frontal 
Convolution of Left Hemisphere (fig. 27). 


Troglodytes niger, No. xiv., young adolescent {%, strong, tame. 
Very vociferous; subject to fits of anger and excitement, and then becomes 
very noisy, uttering a considerable variety of sounds, scolding, greeting, 
petulant, etc. On April 9, after animal had been in laboratory for seven 
weeks, the skull was trephined under deep anesthesia, and the lower 
region of left hemisphere was exposed and stimulated by unipolar faradisa- 
tion. The post-centralis in the field laid bare, i.e. up to inferior genu, was 
nowhere found excitable; precentralis was excitable continuous throughout 
its exposed length, i.e. up to genu inferius. The excitable field faded off 
somewhat gradually in the frontal direction. Certain of the cortical points, 
the responsive movements of which were specially observed, were topo- 
graphically recorded as given on the map (fig. 27). A field of cortex in 
front of the lower end of gyrus precentralis was also mapped (fig. 27) 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 205 


and explored by faradisation from point to point. It nowhere yielded any 
regularly recurring results as far as could be seen except, in upper and fore 
part, opening of eyes; in most parts of it excitation was not followed by 
any visible result whatever. The larynx was kept under laryngoscopic 
examination for the greater part of the time that this area was being 
explored by the electrode. From points in the precentralis behind this area 
it had been ascertained by laryngoscopic examination that faradisation 
evoked regularly adduction of the vocal cords; the topography of those 
points is recorded in the map (fig. 27). From the field explored forward of 
the precentralis, however, no movement of vocal cords, larynx, lips, or 


Fic. 27.—Map of the left hemisphere of chimpanzee 14; a number 
of the responses obtained to faradisation both at the time of the 
ablation-operation and at the subsequent final exploration are 
mapped. The shaded area limited by sulci and dotted lines shows 
the ablated area of the cortex. Ablation-experiment 7. 


tongue was elicited. The larger portion of this latter field was then 
ablated to a depth of about a centimetre and for the area enclosed by 
the dotted line in fig. 27. There was little hemorrhage. The dura was 
stitched together, and the skin wound closed. Aseptic precautions had 
been adopted throughout the operation. 

On recovering from the operation narcosis the animal showed no facial 
or other paresis, and no impairment of its vocalisation was detected. It 
was excited somewhat after coming round from the anesthetic, and uttered 
a variety of vocal sounds. The pitch of its voice was thought to be rather 
higher than had been usual, but this may have been an after-result from 
the inhalation of the anesthetic, and was not observable later in the after- 
noon nor subsequently. 


Next day: “animal seems irritable; he screams, and is more noisy than 


206 Leyton and Sherrington 


usual on having to return to his cage after feeding and playing. Is finally 
pacitied on being given more grapes than customary allowance. Is certainly 
as vociferous as ever; seems to employ quite as wide a range of various 
sounds as before. Calls and shouts when alone or when watched from a 
distance.” 

April 12.—Certainly not the slightest reduction of animal’s vocalisa- 
tion, nor any apparent change in it. Appetite good. Wound healing well; 
but animal seems more excitable than before the operation. 

April 14.—Same condition; still very excitable; wound healing well. 

April 16.—Same as before; wound nearly healed. 

April 17.—Animal has torn off its dressing, and torn up skin flap; 
some hemorrhage. Re-dressed. 

April 20.—Wound infected: animal killed under chloroform. 


Remarks on Foregoing Ablation-Experiments. 


Owing to their lesser remoteness from human type it seems more 
possible in regard to the anthropoid than to monkeys such as macaque 
to infer the animal’s mental attitude at various times. A point which 
impressed us repeatedly was the seeming entire ignorance on the part 
of the animal, on its awakening from an ablation-experiment, of any dis- 
ability precluding its performance of its willed acts as usual. Surprise 
at the failure of the limb to execute what it intended seemed the animal’s 
mental attitude, and not merely for the first few minutes, but for many 
hours. It was often many hours before repeated and various failures to 
execute ordinary acts contributory to climbing, feeding, etc., seemed to im- 
press gradually upon the animal that the limb was no longer to be relied upon 
for its usual services. The impression given us was that the fore-running 
idea of the action intended was present and as definitely and promptly 
developed as usual. All the other parts of the motor behaviour in the 
trains of action coming under observation seemed accurate and unimpeded 
except for the réle, as executant, of the particular limb whose motor cortex 
was injured. And there seemed to be, and to persist for some time, a mental 
attitude of surprise at the want of fulfilment of that part of an act which 
had been expected to occur as usual. The surprise seemed to argue 
unfulfilled expectation, and defect in the motor execution rather than in 
the mental execution of the act, raising the question whether the function 
of part of the cortex ablated in such cases be not indeed infra-mental. 

We would not by this suggest that the part of the cortex in which the 
motor zone is situate may not be involved in processes of synthesis of 
sensation as well as in that of motor and postural action. The recent 
experiments by Dusser de Barenne (11), by minutely localised application 
of strychnine to the cortex of the “motor” zone as well as to other adjoining 
parts of the cortex, clearly give grounds in support of the view that the 
cortex of the motor zone influences sensation. 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 207 


The paresis of the limb whose corresponding motor cortex area had 
been heavily damaged by ablation was severe, as evidenced by imperfection 
of willed movements attempted to be executed by it in the early days 
following upon the inflicting of the lesion. But this paresis was largely 
temporary. Improvement in the willed actions of the limb set in very 
early, and progressed until the limb was finally used with much success for 
many purposes even of the finer kind. Thus after destruction of the 
greater part of the arm areas of both hemispheres the two hands were 
freely and successfully used for breaking open a banana and bringing the 
exposed pulp of the fruit to the mouth. And again, after considerable 
destruction of one leg area the foot was successfully used for holding on 
the bars when climbing about the cage. As we said in our preliminary 
communication, the absence of recrudescence of the hand paresis on ablating 
the remaining intact part of the arm area showed that that latter part of 
the cortex had not taken over the functions, at least not to any marked 
extent, of the ablated portion. “In accord with the absence of recru- 
descence of the hand paresis on ablating the remaining intact part of arm 
area was the finding that faradisation of that part (elbow and shoulder) 
provoked as usual movements of elbow and shoulder, but not of hand 
itself, or only of hand late in a general arm movement, and that very 
rarely. In short, neither the ablation or excitation methods gave any 
evidence that the remaining part of the arm area had taken on the fune- 
tions of the ablated hand area. Neither was the gyrus centralis posterior 
appreciably altered under exploration, and had not become a stimulable 
area for arm, hand, or other movements.” And recently it has been found 
by T. Graham Brown and one of us, and by the former in independent 
observations, that subsequent ablation of the adjoining centralis posterior 
does not cause recrudescence of the arm paresis. Further, as pointed out 
in our preliminary communication, the double arm area lesion showed 
clearly that the regaining of ability to use the limb could not be attributed 
to the arm area of one hemisphere taking over the functional powers of 
the arm area of the other hemisphere after the latter’s ablation. This 
confirms for the anthropoid the result obtained in the dog by Frangois 
Franck (15), and is itself confirmed by an experiment on the chimpanzee 
published by T. Graham Brown and one of us (7) much more recently. 
On the other hand, that in the movements of some parts the motor 
cortex of one hemisphere is supported in its function by the correspond- 
ing part of the motor cortex of the other hemisphere is indicated by our 
ablation-experiment on the eye-closure area. In it the ablation of the 
area from one hemisphere produced very little paresis of the movement, 
but a rapidly successive ablation of the corresponding area from the other 
hemisphere brought about distinct paresis. In this instance the movement 
impaired tends usually to be a bilateral one: and that seems the main 
factor accounting for the different result. 

The absence of obvious symptoms resulting from destruction of a large 

VOL. XI., NO. 2,—1917. 14 


208 Leyton and Sherrington 


part of the left inferior frontal gyrus in a very vociferous chimpanzee has 
probably not much weight in regard to the possible functions of that con- 
volution in man. The experiment and its negative result were mentioned 
in our preliminary communication, at a date prior to the interesting and 
important controversy as to the functions of that gyrus which has led to 
so much recent inquiry in regard to human material. 

As regards the secondary degeneration in bulb and cord, they show that 
the pyramidal tract in the anthropoid (chimpanzee) more closely resembles 
the human than does that of any other animal so far examined. In the 
chimpanzee as in man there is a well-marked uncrossed ventral column 
bundle belonging to the tract, and as in man so in the chimpanzee, to judge 
from our experiments, though they are few, much individual variety exists in 
the relative size of that bundle to the rest of the tract (13). The uncrossed 
ventral column bundle shows degeneration after arm area lesions as well as 
after leg area lesions, but in the latter case its degeneration is traceable 
into the lumbar region, whereas in the former it ceases much higher up 
the cord, although there it may be large. The degeneration at the region 
of the pyramidal decussation shows in addition to the main mass of fibres 
crossing to the contralateral lateral column a small number of fibres sloping 
backward towards the ipsilateral column, as has been shown for the smaller 
monkeys (25, 41, 43), and presumably holds also for man. It is this 
uncrossed pyramidal tract slip entering ipsilateral lateral column which 
probably accounts for the scattered slight degeneration in the pyramidal 
tract area of the lateral column of the cord ipsilateral with the cortical 
lesion, an ipsilateral degeneration observed in all of our experiments. The 
pyramidal-tract degeneration after the arm area lesions was traceable to 
much below the brachial enlargement (cf. Sutherland Simpson and 
W. A. Jolly (48)), but did not reach the lumbo-sacral. In the grey matter 
of the ventral grey horn of the side contralateral to the cortical lesion a 
heavy degeneration in the minute fibres was evident, in the brachial 
segments after arm area lesion, in the lumbo-sacral enlargement after leg 
area lesion. 


IV. EXPERIMENTS ON GYRUS CENTRALIS POSTERIOR. 


Our experiments on this portion of the cortex may be divided into two 
groups :— 

1. Observations by stimulation. 

2. Observations by ablation. 


1. Results of Stimulation. 


Faradism, applied to the free face of centralis posterior in the same manner 
as to gyrus centralis anterior, although readily evoking motor responses from 
the latter, failed to excite in a similar way any detected motor responses 
from gyrus centralis posterior in chimpanzee, orang, or gorilla. In all our 
experiments this experiment was made, and in all the same negative result 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 209 


was obtained. In many experiments the whole length of post-centralis was 
systematically explored, in some only a part of it, namely, that corresponding 
with face area or arm or leg area of the precentralis. Stronger faradisa- 
tion of post-centralis also failed to give motor responses ; indeed frequently 
the strength of faradism applied to post-centralis was carried far beyond 
the strength ordinarily permissible for reliable physiological observations, 
and still quite failed to evoke any detectable effect. 

Faradism of the cortex of the centralis posterior convolution, though 
not itself eliciting movement, does, however, when employed at certain 
places, facilitate sometimes, as was mentioned in our preliminary communi- 
cation, the elicitation of movement by faradisation at certain points in 
about the same horizontal level of the precentral convolution. In other 
words, from certain parts of the post-central gyrus a facilitating influence 
can be exerted upon somewhat adjacent parts of the precentral gyrus. 
This observation has been confirmed for the human brain by C. K. Mills 
(27, 28, 29). In our observations on the anthropoid we met the phenomenon 
especially in the region below the inferior genu; for instance, stimulation 
of a point of centralis posterior close to the central sulcus facilitated 
response of a point opposite to it in precentral gyrus yielding movement 
of the contralateral side of the lips. 

Under certain special circumstances faradism may at times evoke, as 
was mentioned in our first preliminary communication, reactions from the 
post-central gyrus itself, though the conditions are sufficiently different 
from those which obtain for elicitation of responses from the centralis 
anterior to exclude the centralis posterior being accepted as equivalent 
to centralis anterior cortex. When the centralis posterior near to the 
central fissure is faradised immediately after elicitation of a motor response 
from centralis anterior at a point in the latter lying about opposite the 
point faradised in centralis posterior, the motor response obtained from 
the centralis anterior may reappear, and this even a few times in succes- 
sion, though not for many unless centralis anterior be restimulated. This 
“ echo-response ” is a phenomenon of considerable constancy. Our observa- 
tions on it were made chiefly in the region of the inferior genu and below 
that, and with motor responses in lips, thumb, or index finger. Graham 
Brown (4, 5)! has, independently of us, observed the phenomenon in 
regard to flexion of the arm, and in small monkeys macacus and cerco- 
pithecus as well as in chimpanzee. We met with it in all of the three 
anthropoid apes. We have been inclined to regard it as analogous to a 
phenomenon met with along the anterior confines of the motor region. 
The anterior limit of the excitable region as examined by faradism seems 
to merge somewhat graduatim into the inexcitable surface beyond it. 
Towards the end of a lengthy stimulation-experiment, when the region 
has been repeatedly stimulated at many points, the anterior limit of it 
seems not rarely to extend forward farther that it had done at first. Re- 

1 Journ. of Physiol., 1914, xlviii. p. xxx ; Quart. Journ. of Exper. Physiol., 1915, ix. 82. 


210 Leyton and Sherrington 


iterated faradisation of a point, e.g. close in front of one regularly yielding 
thumb movement or finger movement, will ultimately sometimes yield that 
movement, especially if faradised directly after the point yielding it regu- 
larly has responded. The posterior boundary of the motor area lies buried, 
as we have shown, within sulcus centralis. In some specimens, and in 
some parts of the length of the sulcus, the limit seems to correspond pretty 
exactly with the floor of the fissure; but it seems to vary, and in some 
specimens it les up the posterior wall of the fissure. Points on the free 
face of the post-centralis and near to the lip of the fissure are, therefore, 
not very distant from the posterior limit of motor area itself. 

In two cases, one of them including both hemispheres, we ablated the 
arm area back to sulcus centralis, and in none of these, neither immediately 
after the ablation nor subsequently at periods amounting in the longest case 
to four months from the ablation, were we able to evoke motor responses 
by any means from the face of centralis posterior for that length of it 
corresponding with the centralis anterior ablation. In two cases we 
ablated that portion of leg area which lies on the external face of the 
hemisphere back to sulcus centralis, and in those cases also neither immedi- 
ately after the ablation nor twenty-one and thirty-three days later re- 
spectively did faradisation of the corresponding portion of gyrus post- 
centralis evoke any motor response. In one case (ablation-experiment 8) 
we ablated the facial area of precentralis back to the sulcus centralis, and 
in that case likewise faradisation failed to elicit either immediately after 
the ablation or twenty days later any motor response from the correspond- 
ing region of centralis posterior. 


2. Ablation. 


We have made three experiments (ablation-experiments 9, 10, 11) on 
ablation of gyrus post-centralis, all partial, all on the chimpanzee, and all 
in the left hemisphere. In one the gyrus was removed with the knife from 
opposite the superior genu above to a little below the inferior genu below. 
The anterior limit of the ablation was sulcus centralis itself, and the whole 
width of the gyrus was removed. Prior to the ablation the gyrus was 
faradised from point to point systematically in the usual way, and yielded 
no motor or other detected results. 

On recovering from the operative chloroformisation the animal became 
somewhat excited, and observation of its movements revealed no paresis; 
it climbed about the cage apparently with normal facility, and picked up 
grapes and other food with the right hand as with the left, and carried 
the food to its mouth seemingly equally well with either hand. Nor was 
there any obvious sign of asymmetry of the face or paresis of the lips or 
eyelids, No abnormality of skin sensitivity was detected, but such ex- 
amination was difficult, as the animal was not tame. 

A week later, no paresis having in the meantime been manifest, the 
animal was again chloroformed, and the whole of the precentral gyrus, 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 21] 


systematically examined by the electrodes, revealed nothing abnormal in 
its responses, either in the direction of excessive or of defective response. 
The animal was then destroyed by chloroform. 

In another experiment (fig. 28) the lower part of the gyrus was exposed, 
and after point-to-point examination of it with the electrodes had been 
found to yield no motor or other detected responses the gyrus was ablated 
with the knife from close above inferior genu downward for one-half of its 
length below that genu. The anterior boundary of the ablation was made 
by thrusting the knife between the lips of that part of sulcus centralis and 
sloping it downward and backward through the posterior wall of the fissure 


Fic, 28.—Lesion in gyrus centralis posterior giving no symptoms and no degeneration in bulb 
or spinal cord ; chimpanzee, ablation-experiment 10. 


thus avoiding the floor of the sulcus and the deepest portion of the posterior 
wall. The width of the ablated portion in its upper part extended back to 
the lower spur of the post-central sulcus, and below that parallel with sulcus 
centralis and 15 mm. behind it. The lowest boundary ran horizontally from 
sulcus centralis to the posterior cut behind. 

On recovering from the anesthetic the animal showed no trace of 
paresis either in hand or face, no impairment in mastication or drinking 
or swallowing, in short, no detected departure from normal motility. The 
skin of the right half of the face could feel, but it was not possible to 
assure oneself that its sensation was undisturbed and actually normal. 
Fifteen days later the animal was used for systematic examination of the 
motor cortex, and the precentral gyrus throughout its facial as well as its 


212 Leyton and Sherrington 


other regions yielded responses in which no abnormality was detected on 
the left side any more than on the right. The bulb and cord were 
subsequently sectioned and examined by the Marchi method for degenerat- 
ing tracts as far forward as the upper half of the pons. The pyramidal 
tracts showed no degeneration, either in pons, bulb, or cord. 

A third experiment, similar to the preceding, but with a lesion extend- 
ing farther down the centralis posterior, resulted similarly, no obvious 
etiects being detected. We tried the lower part of the gyrus by preference, 
because it is in that part that previous experimenters placed extensive 
motor centres. 

V. THE INSULA. 


In one chimpanzee we exposed the insula for the larger portion of its 
extent, and tested its surface by faradism, both by the unipolar and the 
bipolar method. To expose it, the lower portion of gyrus centralis anterior, 
which had been examined by stimulation and given the usual results, had to 
be removed. From the whole surface of insula tested we obtained no detect- 
able results, although the gyrus centralis anterior where not removed con- 
tinued at the time to respond readily. Two fissures exposed in the insula 
were also opened up and the electrode applied, but no result was elicited. 


VI. THE THRESHOLDS OF FARADIC EXCITABILITY OF THE MOTOR CORTEX 
oF Cat, MacaquE MONKEY, AND CHIMPANZEE COMPARED. 


We made some observations in regard to this by a procedure for 
the suggestion of the electrical arrangement of which we are indebted 
to Professor J. S. Macdonald. Cat, macaque (Macacus sinicus), and 
chimpanzee were anzesthetised by chloroform and ether mixture in the 
usual way. In each animal the motor cortex was then exposed suitably 
for stimulation of the fore-limb area. A copper plate with binding screw 
was applied over a pad of cotton-wool soaked in strong salt solution to the 
shaved skin of the hind-leg in each animal (fig. 29). The copper plate 
applied to one of the animals (A) was attached by a wire to one end of the 
short-circuiting key in the secondary circuit of the physiological induc- 
torium. Another wire attached the binding screw on the handle of an 
electrode of the pattern (stigmatic) used by us for unipolar faradisation to 
the copper plate on a second of the animals (B), and to the copper plate 
attached to the third animal (C) a similar electrode was similarly attached. 
Finally, to the short-circuiting key on its side opposite that connected with 
the copper plate on animal (A) a wire attached a third electrode of similar 
pattern to the other two. The secondary circuit's short-circuit key being 
closed, three persons in separate charge of the three animals, and each 
controlling one of the stigmatic electrodes, applied his electrode to a spot 
in the “arm area” of cortex, the spot being in each case one which had 
been found to elicit flexion of elbow. The three animals were thus 
connected in series in the secondary circuit of the inductorium beyond the 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 213 


short-cireuiting key. The stigmatic electrode in each animal was that 
which was cathode at the break-induced current. The three electrodes 
being applied, the primary current was then started by closing its key, and 
the vibrating spring, about 40 per second, operated in the primary circuit. 


Fic, 29.—Scheme of faradisation for testing the relative excitability of the motor 
cortex in cat, macaque monkey, and chimpanzee ; see text. 
The short-circuiting key in the secondary circuit was then opened. The 
observation consisted in finding at what distance, as the secondary coil was 
brought nearer to the primary, one or other animal gave a motor response, 
and whether response was given by one or other animal under distinctly 
weaker stimulation than by the other animals, 


EXPERIMENT, APRIL 20. +, response (elbow flexion). 0, no response. 
2 See eee 
Distance of | 

2nd coil from Cat. 

primary in cm. 


Chimpanzee, Macaque. 


| 13 
12 
11 
10 
95 
9 
85 
9 
9°5 
10 
10°5 
11 
115 
12 
12°5 
13 


The results we obtained are illustrated by the above protocol. It will 
be seen that there was no clear indication that response was elicited by 


ooo¢4t+++4+4++4++000° 
coot4t4+4+4+4+4+4+4+4+4+ 000 
cooot¢tHt+444+4+4++4+000 


214 Leyton and Sherrington 


weaker stimuli in any one of the three animals than in the others. On 
the contrary, the evidence was that the threshold of excitability was closely 
similar for all three. 


VII. INFLUENCE oF LocaL CoLD AND WARMTH APPLIED TO THE SCALP 
ON THE LocAL TEMPERATURE OF THE CORTEX UNDERNEATH. 


Large chimpanzee, anesthetised and trephined in lateral frontal region 
of left side; the thickness of skull in this place was 3°5 mim. as measured 
on the edge of the button of bone removed. The trephine hole was 
slightly enlarged antero-posteriorly, the dura opened, and the bulb and 
stem of a slightly curved and flattened small thermometer was introduced, 
and gradually shifted under the dura for 4°5 cm. in a posterior direction. 
The bulb thus lay under dura and bone about 4°5 cm. behind the hole in 
the skull and in a postero-lateral direction from it. The issuing stem of 
the thermometer, wrapped in dry cotton-wool for its part next the lips of 
the wound, was readable. The room temperature was 27° C. After the 
intracranial thermometer had been giving steady readings for a time an 
ice-bag was applied to the scalp in the left parietal region, approximately 
over the seat of the bulb of the thermometer inside. The diameter of 
application of the bag was 9 cm. Its edge did not come quite close to the 
edge of the skin wound and the emergent thermometer. 

The following illustrates the observations obtained. 


Time. sas ee Rectal temp. Pulse. 
emp. 
2.30 36°4° C. 36°2° C. 74 
2.33 36°4 
2.35 364 
2.36 ice-bag applied 
2.38 35°9 
2.39 35°1 
2.40 34°4 
2.41 34:0 
2.42 33'8 
2.43 33°6 
2.44 33°4 
2.45 33°2 
2.46 33°1 
2.47 331 36°4 75 
ice-bag removed 
2.48 331 
2.49 33°2 
2.50 33°7 
2.51 34°] 
2.52 34:5 
2.53 34:9 
2.54 35°2 
2.55 35°5 
2.56 35°7 
2.57 35°8 
2.58 35°8 
3.0 36:0 36:0 75 | 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 215 


Intracranial 


temp Rectal temp. Pulse. 


Time. 


Heat applied—water in bag at 55° C. applied over 
same area—to same hemisphere. 


| 
3.30 35°9 36°0 
3.35 359 | 
warm bag applied 
3.36 36°3 | | 
3.37 37 
3.38 37°4 
3.39 a7T°0 
3.40 38°0 
3 4] | 38°3 | 
3.42 | 38°5 fresh water at 55° in|to bag. 
3.43 38°6 
3.44 | 38°5 
| bag removed 
3.45 | 38°3 
3.46 | 38-0 i 
3.47 37°3 | | | 
3.48 oil 
3.49 369 
3.50 36°8 


The thickness of the wall of the parietal bone overlying the thermo- 
meter bulb was 3 mm. in fresh state, i.e. about the same thickness as of an 
ordinary human skull in that region. Also a certain amount of temporalis 
muscle intervened between the scalp and bone at the place of application 
of the bag. 

The above instances are representative of the results obtained in the 
short series of observations made. The chimpanzee used in the above 
observations was chosen because, from previous operations on the other 
hemisphere, it was recognised to have a skull of more than average 
thickness. 

From the above, it is seen that by local application of cold and warmth 
to the scalp under conditions similar to their application clinically in man 
the temperature at the cortex of the underlying brain may be veried 
readily over a range of about 10°F. (e.g. 91°5° F-101°5° F.). 


VIII. Errect oN THE EXCITABILITY OF THE Motor CorRTEX OF 
CLOSING THE CAROTID ARTERIES. 


We have observed upon six chimpanzees the effect of closure of the carotid 
arteries upon the excitability to faradism of the hand, leg, and face areas 
of the cortex. In three of these simultaneous closing of the two common 
carotid arteries extinguished the excitability of the cortex. In one of the 
three animals the excitability of the cortex lapsed in 100 seconds in the 
right motor area and in 115 seconds in the left, after simultaneous closure 
of both carotids. The vessels were then released, and the excitability, 
as tested in hand area, returned in 75 seconds in left hemisphere and in 


216 Leyton and Sherrington 


85 seconds in right hemisphere. In this animal closure of the right carotid 
alone diminished the motor responses in right hemisphere practically to 
extinction, though less speedily than did closure of both vessels, and so 
similarly did closure of left carotid those of left motor region. But in the 
second animal closure of one carotid alone did not abolish the motor 
response in either hemisphere, although the closure of both carotids 
together extinguished all motor responses from both hemispheres in 
120 seconds. The responses in this case became re-elicitable on each side 
in 90 seconds after freeing the carotids. In the other case the closing of 
the carotids extinguished cortical excitability only after 4 minutes. No 
convulsions were evoked by any of the occlusions, but transient increase 
of exitability of the cortex was noted in one instance: cf. L. Hill (21). 


IX. FUNCTIONAL GROUPING OF PYRAMIDAL-TRACT FIBRES IN 
CRURA AND PONS. 


The size of the pyramidal tract in the anthropoids is large enough to 
offer a better chance than in animals which are smaller or in which it is 
less developed for testing by faradisation the degree to which the various 
fibre groups from the various fields of the motor cortex lie separate or 
commingled in the tract at various levels. In the largest of our orangs 
we removed the whole brain in front of a transection through the posterior 
part of the anterior colliculi, and examined by faradisation the cross- 
section of the crusta. As so exposed, the pyramidal-tract fibre bundles 
run of course perpendicular to the plane of the transection. With fibres 
thus exposed the unipolar method of faradisation gives better opportunity 
than does the bipolar for minutely localised stimulation. With the former 
method the current lines converge in a direction more nearly parallel with 
the lengthwise direction of the fibres it is devised to excite. Examined 
by unipolar faradisation, the results obtained from the orang’s crusta were 
as follows:—The most lateral third of the cross-section gave no detected 
responses at all, neither did the most mesial fourth. The intermediate 
portion gave responses which, taken in sequence from its lateral edge to its 
mesial, were in the following order: toes, ankle and knee, hip, trunk, arm, 
face, and tongue (fig. 30, A). There was very great overlapping of the 
areas yielding these results; thus it was easy to obtain from some points 
concurrent movement in leg, trunk and arm, or again of arm, face, and 
tongue. 

The severity of the operation necessary for exposing such a cross- 
section did not allow a repetition of the observations at a lower level in 
the same animal. But in the largest of our gorillas we removed the whole 
brain in front of a transection through the highest part of the pons, and 
examined by faradisation with the unipolar electrode the cross-section of 
the pyramidal tract at that level. The results obtained in both right and 
left pyramidal tracts were similar, confirming each other. They were that, 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 217 


although facial, lingual, brachial, and hind-limb movements of the contra- 
lateral side were evoked together by stimulation anywhere in the cross- 
section of the tract at that level, the movements in face and tongue 
predominated greatly when the electrode was applied to the mesial portion 
of the cut face of the tract, and movements of toes when the application 
was to the outer lateral portion of the tract, while, when the electrode lay 
about midway between the mesial and lateral borders of the tract, there 
was marked predominance of the finger movements (fig. 30, B). The 
inference is therefore that although by that level the pyramidal-tract 
fibres from face, arm, and leg areas have become a good deal commingled 


| 
o/ 


es AN 


Fic. 30.—A, outline of cross-section of the crura cerebri in a large orang, to show the position of the 
spots in the crusta whence unipolar faradisation elicited separable movements of opposite side. 
1, movement of toes and ankle; 2, hip and knee ; 3, abdominal wall and chest wall ; 4, fingers, thumb, and wrist ; 
5, face and tongue ; between 3 and 4, movements of elbow, wrist, and shoulder. 
B, outline of cross-section of anterior part of pons of gorilla, to show points whence move- 
ments predominating in toes (¢), in fingers and thumb (/), and in face (F) respectively were 
elicitable by unipolar faradisation. 


those for face predominate toward the mesial side of the tract, those for 
leg towards the lateral, and those for arm in the middle part of the tract’s 
cross-area. 

Along with these observations may be mentioned a feature observed in 
the pyramidal-tract degeneration following on destruction of the arm area 
in the chimpanzee (v. sup., p. 192, figs. 20, 22). In the cross-section of the 
degenerated pyramid the degenerated fibres are somewhat less numerous 
in the ventrolateral part than elsewhere. 

And the same cases showed that some of the fibres from arm area are 
among the highest of the fibres which decussate to the contralateral side 
in the pyramidal decussation. 

In another of the orangs we made the following observations :—After 
the hemispheres had been systematically explored with the electrode, and 


218 Leyton and Sherrington 


were still responding well, the spinal cord was exposed at the 4th thoracic 
segment, and the right lateral half carefully severed. Subsequent micro- 
scopic examination of the semi-section proved it to have been an accurate 
one, the whole right half being severed, with a slight trespass only into 
the left side in the dorsal column. Stimulation of the left cortex evoked 
after this lesion unaltered responses from face and arm area, but no 
response at all from trunk area or leg area. Responses from right cortex 
as before the semi-section. A second right side semi-section of the cord 
midway between 3rd and 4th cervical roots was then made. Microscopie 
examination subsequently showed that in this semi-section the mesial part 
of right ventral white column escaped severance. Stimulation of the left 
hhemisphere’s face area after this second semi-section evoked facial move- 
ments as before, but stimulation of the arm area evoked no intrinsic arm 
movements, although responses in trapezius and rhomboids were obtained 
from it. Responses from right hemisphere remained unaltered. 


X. SUMMARY OF CONCLUSIONS. 


1. The “motor” area of the cortex in the three species of anthropoid 
examined (chimpanzee, orang-utan, and gorilla), as determined by faradisa- 
tion, embraces almost all of the free surface and a large part of the sulcal 
surfaces of gyrus centralis anterior; it also extends over the mesial border 
upon gyrus marginalis for a distance about half-way toward sulcus cinguli, 
in agreement with Campbell’s delimitation of his “precentral type” of 
cortex in chimpanzee and orang. 

2. The proportion of motor area buried in the sulci is probably usually 
about one-third of the whole area. 

3. Although the broad “localisation” of the responses of the various 
main motor parts of the opposite half of the body follows a well-fixed 
topographical scheme in this cortex, the minuter localisation, as examined 
by faradisation, is subject to temporal instability. 

4, This instability is largely the expression of mutual influences exerted 
transiently by the physiological states for the time being of different points 
of the motor cortex, and of the sub-cortical centres they connect with, one 
upon another. These influences make themselves felt as “deviation of 
response,” “reversal of response,” and “ facilitation,’ phenomena all seem- 
ingly akin. 

5. Subject to this temporal instability, details of localisation of various 
movement groups in chimpanzee, orang, and gorilla are described. Difter- 
ences in the smaller details of the localisation were met with from in- 
dividual to individual of the same species, and between the right and left 
motor areas of the same individual. Making all allowance for experi- 
mental error, these differences, in some particulars, seem too large to be 
accounted for fully by that or by temporal instability of the cortex; they 
may represent, as Franz urges for analogous differences he found in 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 219 


macaque, permanent individual differentia existing from hemisphere to 
hemisphere. 

6. The motor responses obtained by faradisation from a baby chimpanzee 

equalled in differentiation, as far as could be seen, the average of those 
obtained from the adult of any of the three anthropoid species examined. 
And “epilepsy” was produced neither more nor less easily than in those. 
7. The anterior edge of the motor area seems to fade away somewhat 
gradually into inexcitable cortex. Farther forward still is a large diffuse 
field, from scattered points of which, in the middle and inferior frontal 
convolutions not extending to their more anterior parts, conjugate devia- 
tions of the eyeballs and opening of both eyes are elicitable. but stronger 
faradisation is required for these, nor are the results so regular as with the 
responses of the motor area proper. 

8. Eyeball movements similar to those just mentioned are likewise 
obtainable from the occipital pole and from the calcarine region. 

9. The motor area for face and tongue movements seems, relatively to 
the rest of the motor area, more extensive in orang than in chimpanzee. 
Apart from that distinction, there seemed no clear difference between the 
motor area from species to species of the anthropoids examined. The 
largest and most highly developed brain we examined was that of a gorilla, 
and the motor area in that specimen appeared to be, on the whole, the 
most extensive and differentiated of those experimented upon. 

10. The motor cortex may be regarded as a synthetic organ for 
compounding and re-compounding in varied ways movements of varied 
kinds of scope from comparatively small, though in themselves well 
co-ordinate, fractional movements. For this synthesis the motor cortex is 
provided with, ie. has at call, these partial or fractional movements and 
postures. The cortex obtains these partial movements, perhaps by analytic 
powers of its own, from the bulbo-spinal mechanisms, but the higher of the 
synthetic results of the bulbo-spinal mechanism exhibit, as judged from cat 
and dog, certain only of the kinds of compound movements which the 
motor cortex gives. From the recomposition of these partial movements 
into wholes of varied pattern and sequence there result motor acts which, 
taken in their entirety, making use of the same fractional pieces, attain 
with them aims of varied scope by varying the spatial and temporal 
combinations of them. 

11. The free surface of gyrus centralis posterior was found to differ 
from that of gyrus centralis anterior in not being similarly excitable by 
faradisation. Faradisation behind the sulcus centralis can, under certain 
circumstances, evoke reactions from the cortex, but these are doubtful 
for acceptance as equivalent to “motor-area” reactions. They appear as 
‘echo-responses” when the faradisation is made to follow directly and 
quickly on faradisation of gyrus centralis anterior in the near neighbour- 
hood, i.e. about the same horizontal level and not far from sulcus centralis. 
The “echo-response ” thus obtained from gyrus centralis posterior repeats a 


220 Leyton and Sherrington 


response just previously given from centralis anterior, and soon dies out 
under repetition of the stimulus to gyrus centralis posterior, unless stimula- 
tion of gyrus centralis anterior is repeated to renew it. 

12. Stimulation of the middle and posterior parts of the inferior frontal 
convolution of left hemisphere failed in chimpanzee, orang, or gorilla to 
evoke any vocalisation. Ablation of a large portion of that area in one 
chimpanzee, chosen because it was a noisy and vociferous animal, produced 
no obvious impairment or change in vocalisation. 

13. Faradisation of the surface of the insula failed. to evoke any 
detectable results. 

14. Unipolar faradisation of the cut cross-face of the crusta (orang) 
evoked responses separately in toes and ankle, hip, trunk, arm, and face 
from a series of points taken in order from without inward (mesially). 
From the cut cross-face of the pyramidal bundles in the pons (gorilla), 
unipolar faradisation evoked toe movement predominantly from the most 
lateral, face-tongue movement predominantly from the most mesial, and 
finger movements predominantly from the intermediate. 

“15. Ablation of the cortex of the larger portion of an arm or leg area 
in gyrus centralis anterior produced heavy paresis of the corresponding 
limb, but this paresis quickly lessened, and the limb soon regained in large 
measure its volitional motility, and became successfully used for climbing, 
picking up food, picking the teeth, ete. Ablation further of the greater part 
of the arm area of the second hemisphere, after previous ablation of the 
greater part of that area from the other hemisphere, induced no recrude- 
scence of paresis in the already paretic and partly recovered arm. After 
the double lesion considerable recovery of the volitional use of both limbs 
somewhat rapidly ensued, the hands, for instance, being used freely in 
climbing, picking up food, ete. 

16. The degenerations in the spinal cord following on limb-area lesions 
exhibited a large crossed pyramidal tract, extending more to the edge of the 
lateral column than in man, and in this respect resembling a feature seen in 
the macaque cord. There was obvious also a slight ipsilateral pyramidal 
tract in the ipsilateral lateral column, derived from a small portion of the 
fibres of the pyramid passing not to the lateral column of the crossed side, 
but to that of the ipsilateral side. There was also evident in two of the chim- 
panzees an ipsilateral ventral pyramidal tract similar in position and relative 
size to that (“direct Py. T.’) commonly seen in man; this is not existent in 
the macaque. The pyramidal-tract degeneration after the arm-area lesions 
descended beyond the brachial enlargement of the cord, but did not reach 
the lumbo-sacral enlargement. The pyramidal-tract degeneration ensuing 
on the leg-area lesions descended the whole length of the cord. Many fine 
degenerate fibres (collaterals) were visible in the ventral horn of grey 
matter among the motor perikarya on the side contralateral to the cortical 
lesion in the brachial segments of the arm-area lesion and in the lumbo- 
sacral enlargement in the leg-area lesion. 


The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 221 


17. Ablations of portions of the free surface of gyrus centralis posterior 
gave rise to no obvious symptoms of paresis, nor, in the one case whose bulb 
and spinal cord were examined, to degeneration of the pyramidal tract. 

18. The threshold of faradie excitability of the motor cortex, as tested 
in the arm area, seems to be practically similar in cat, macaque, and 
chimpanzee. 

19. In some chimpanzees occlusion of the two carotid arteries renders 
the motor cortex inexcitable to faradism, and does so rapidly, e.g. 2 minutes. 
After release of the arteries, responses to faradic stimulation reappear in 
about 14 minutes. In one animal occlusion of one carotid alone reduced 
the excitability of the motor area of the corresponding hemisphere almost 
to extinction, but in another animal occlusion of one carotid alone did 
not markedly depress the excitability. The anthropoid brain, unlike the 
brain of the smaller monkeys, has frequently a Circle of Willis of human 
pattern (20). 

20. In a chimpanzee with a cranial vault of about the thickness of the 
average human, the application of local cold (ice-bag) and warmth over the 
parietal scalp rapidly attected the temperature of a thermometer bulb lying 
under the dura against the cerebral surface beneath the region of applica- 
tion of the local cold or warmth outside. 


It is a pleasure to express here our thanks to Professor Harvey 
Cushing of Boston, to Dr A. W. Campbell of Sydney, and to Dr Alfred 
Froéhlich of Vienna for valued co-operation and kind assistance in many 
of the experiments. To Dr Besredka, Dr Weinberg, and the late 
Professor Metchnikoff of the Institut Pasteur we are indebted for one 
of the orangs used. We have much pleasure too in recording our acknow- 
ledgment of the energy of Mr E. G. Cox, assistant in the laboratory, in 
obtaining the animal material, and of his skill and care in all matters 
pertaining to the management of it. 


LIST OF REFERENCES. 


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(3) GraHam Browy, T., this Journal, 1915, ix. 81; ibid. 101; ibid., 117; 
ibid., 131. 

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222 The Excitable Cortex of the Chimpanzee, Orang-Utan, and Gorilla 


(8) CampBELL, A. W., Histological studies in the localisation of cerebral func- 


tion, London, 1905. 

(9) Deserine, J. et THomas, Arch. de physiologie normale et pathologique, 
Paris, 1896. 

(10) Dxgsenine, J., Presse médicale, Paris, 1906. 

(11) Dusser DE Barenne, J. G., this Journal, 1916, ix. 355. 

(12) Ferrier, D., Functions of the Brain, London, 1876. 

(18) Frecusic, P., Leitungsb. in Gehirn und Riickenm, Leipsic, 1876. 

(14) Francz, E. P., Phil. Trans., London, B, 1889. 

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CARVILLE et Durst, Archives de physiologie, Paris, 1875, 437. 

(16) Franck, Frang., et Pirres, Gazette médicale, Paris, Mars, 1880. 

(17) Franz, SHEPHERD I., Psychological monographs, 1915, xix., No. 1, 80. 

(18) Frouuicu, ALFRED, and C, S. SHerRrineToN, Journ. Physiol., 1901, xxviii. 14. 

(19) Grtnspaum, A. S. F., and C. S. SHerrineton, Proc. Roy. Soc., 1901, Ixix. 
206; ibid., 1902, : 

(20) Grunspaum, A. 8S. F., and C. S. SHerrineton, Brain, 1902, vol. xxv. 270. 

(21) Hixt, L., The circulation in the brain, London, 1903. 

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xxvil. 63. 

(23) Kennepy, R., Phil. Trans., London, 1914, B, ccev. 27. 

(24) Marte, P., Semaine médicale, 1906, Paris, Oct. 17, Nov. 28. 

(25) Mguuus, E. L., Proc. Roy. Soc., 1894, lv. 208. 

(26) Metiuvs, E. L., Proc. Roy. Soc., 1895, lviii. 206. 

(27) Mitts, C. K., Proc. Philadelph. Cong. Med., Sept. 30, 1904. 

(28) Mitts, C. K., and C. H. Frazier, Med. Bulletin, Univ. of Pennsylvania, 


July, 1905. 
(29) Mitus, C. K., and H. Weisensoroueu, Journ. of Ment. and Nerv. Dis., 


1906, 619. 

(30) von Monakow, C., Die Localisation im Grosshirn, 1914. 

(31) von Monakow, C., Ergebn. d. Physiologie, 1907, p. 334. 

(32) Mort, F. W., E. Scnusrer, and C, 8. SHerrineton, Proc. Roy. Soc., 1911, 
B; ixxxiv. O70. 

(33) Mott, F. W., and E. A. Scudrsr, Brain, 1890, xiii. 164. 

(34) Muratorr, W., Arch. Physiol., 1893. 

(35) Osporne, W. A., and B. Kinvineron, Brain, 1910, xxxiil. 261. 

(36) Roar, H. E., and C. S. SHerrineron, Journ. Physiol., 1906, xxxiv. 315. 

(37) Scudrer, E. A., Text-book of Physiology, Edinburgh, 1900, ii. 747. 

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(40) Saerrineton, C. S., Journ. Physiol., 1889, x. 429. 

(41) SHerrineton, C.S., Lancet, Feb. 3, 1894. 

(42) Suerrineton, C. S., Integrative Action of the Nervous System, New York 


and London, 1906. 
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THE METABOLISM OF VOLUNTARY MUSCLE. I.: THE EFFECT 
OF PROLONGED EXCITATION OF MOTOR NERVES ON 
THE CREATINE CONTENT OF LIMB MUSCLES. _ By 
W. H. TuHompson. (From the Physiology Laboratory, Trinity 
College, Dublin.) 


(Received for publication 21st October 1916.) 


NOTWITHSTANDING the amount of research that has been devoted to a 
study of the changes produced by work in the creatine content of voluntary 
muscles, it cannot be said that the matter is finally settled. 

Of the earlier investigators, Liebig (1847) (1), Sarakow (1863) (2), 
Sezelkow (1866) (3), and Monari (1887) (4) all found an increase of 
creatine after prolonged activity. On the other hand, Nawrocki (1865) (5) 
found no increase in the tetanised muscles of the frog or fowl; while Voit 
(1868) (6) observed a decrease in those of the frog. Sezelkow drew his 
conclusions from one experiment, while Nawrocki’s results were in- 
constant; one of three experiments on frogs showed a marked increase 
in the excited muscle. The methods employed by the earlier workers 
were, moreover, defective. 

The results of recent workers are more uniform. Thus Mellanby 
(1908) (7), von Fiirth and Schwarz (1911) (8), and also Seaffidi (1913) (9), 
could find no change, while Graham Brown and Catheart (1909) (10) 
obtained in isolated frog’s muscle a slight increase after tetanic excita- 
tion. They found, on the contrary, a slight decrease in both the frog 
and the rabbit, with the circulation intact. Lastly, Pekelharing and 
Hoogenhuyze (1910) (11) observed in the rabbit a decrease after section 
of the limb nerves, whereas an increase was seen after section of the 
posterior nerve roots. In the frog a decrease was obtained on tetanisation 
of the limb muscles (deprived of circulation); on the other hand, an 
increase with opening and closing shocks at the rate of 24 per minute. 

The problem has also been attacked from another side, namely, by 
observing the effect of muscular work on the output of creatine and 
creatinine in the urine. 

Of workers on these lines, K. B. Hofmann (1869) (12) observed no 
effect from moderate exercise, while Grocco (1886) (13) found a marked 
increase in the case of soldiers after a strenuous march. Gregor (14) 
also observed an increase after strenuous work, as did Moitessier (1891) 


(144) on a constant diet after long walks. Both these findings were 
VOL. XI., NO. 3.— 1917. 15 


224, Thompson 


confirmed by Hoogenhuyze and Verploegh (1905) (15). After severe 
muscular work, in a condition of absolute fasting, the excretion of 
urinary creatinine was increased. The same was also observed (1910) 
in mountain climbing when the oxygen supply was deficient. Ordinary 
muscular exercise produced no effect. 

Pekelharing (1911) (16), who assigns a special function to creatine 
in connexion with muscular tonus, found an increased excretion after the 
muscles had been kept tonically contracted for long periods—for example, 
in the maintenance of the military position. 

It does not, however, follow that an increased output in the urine after 
muscular work necessarily implies a reduction of creatine in the muscles. 
The contrary may happen, as in starvation. Thus it has been shown by 
Demant (17), and more recently by Mendel and Rose (18), that when 
animals are starved the creatine content of the muscles is increased, at all 
events relatively, notwithstanding a simultaneous excretion of creatine in 
the urine. An absolute increase in the muscles is, however, improbable. 

In studying the direct effects of activity on the creatine of muscles, 
attention has not always been paid to simultaneous variations in the 
content of water. Ranke (19) found an increase of water in tetanised 
muscles. This has been confirmed by many subsequent observers, including 
Danilewski (20), Wortz (21), Ganiké (22), and Barcroft (23). To be 
certain, therefore, of a variation in any of the solid constituents during 
the activity of muscle, it is essential to ascertain the content of water 
in it and allow for alterations if they have occurred. 

In the following investigation this was kept in view. Cats were em- 
ployed, the animals being anzsthetised and decerebrated according to 
Sherrington’s method. Care was taken to prevent loss of heat by 
enveloping the head in cotton-wool and by pinning a layer of felt round 
the body. A hot-water can was also let into the operating table on which 
the animal was placed. 

The muscles of the right leg were removed immediately after decere- 
bration and used for comparison with those of left. The anterior crural 
and sciatic nerves on the left side were divided and armed with shielded 
electrodes after removal of the muscles of the right side. The nerves on the 
left were excited for periods varying from two to two and three-quarter 
hours. The excitation was intermittent, each period of one-minute 
stimulation being followed by a pause of two-minutes rest. 

In removing the limb muscles, a line of stout sutures was laid along the 
flexure of the thigh, passed through the limb from front to back. Before 
tightening these, an elastic bandage was placed on the limb from below up. 
After the sutures were tied and the elastic bandage removed, the skin over 
the thigh muscles was divided and reflected as far down as the heel. The 
muscles were then stripped from off the bones of the thigh and leg and 
the limb amputated at the knee joint. Cotton-wool was then wrapped 
round the femur, and over this the skin replaced. In this way any slight 


The Metabolism of Voluntary Muscle 225 


oozing of blood from the cut surface of the muscles was arrested and loss 
of heat at the same time prevented. 

The muscles after removal were rapidly cleaned of fat and adherent 
connective tissue, care being taken to prevent loss of moisture by keep- 
ing them in a capsule placed under a bell-jar the roof of which was lined 
by a layer of moistened filter-paper. The muscles were then rapidly 
minced, mixed thoroughly, and samples weighed on a delicate balance 
for the different analyses. 

Determinations were made of the content of water, of creatine, creati- 
nine, total nitrogen, and of ash. The samples to be dried were placed under 
alcohol over night, then dried for several hours in a steam-heated oven at 
97° C. Afterwards they were dried to a constant weight at 105° C. In the 
case of those used for determination of the ash, incineration followed drying 
in the steam oven. Creatine and creatinine were determined by Folin’s 
recent method, using as standard a solution of creatinine zine chloride 
recently prepared. The sample of muscle used for creatine was also 
employed for total nitrogen determination. After boiling in the auto- 
clave, the contents of the flask were filtered through a small plug of 
glass-wool into a 200-c.c. flask. The extracted flesh and glass-wool 
were then finely ground up in a mortar and added to the contents of 
the flask. After filling to the mark, the flask was then shaken up 
thoroughly. In this way a uniform suspension of extracted muscle in 
the acid liquid was obtained which could be accurately measured by 
a pipette. After each sample was measured off, the mixture was again 
shaken up before another was taken. The procedure proved satisfactory, 
and gave remarkably concordant results in the duplicate analyses 
carried out. 

In making the creatine determinations, Folin’s directions were followed 
almost exactly. Particular care was taken in adding the 10 per cent. 
NaOH solution both for neutralisation and for development of the colour. 
In all cases this was done by counting the number of drops required, and 
not simply by measurement from a burette. Further, the readings were 
made in groups of four, two from each limb. Thus the muscle extracts 
from the two sides were tested against the same standard solution. 
Possible errors from slight variations in the standard (if made up at 
different times for the two sides) were thus avoided. 

The experiments performed fall into two groups: first, a series in 
which the left anterior crural and sciatic nerves were excited, the blood 
supply of the limb being maintained without interference; and second, a 
series in which the same nerves were excited, but in addition the chief 
artery of the limb was ligatured. This latter was done under the 
impression that possibly some of the products of excitation might remain 
in the muscles and escape being washed away or otherwise removed, as no 
doubt happens when the stream of blood through them is unaltered. The 
results, however, did not conform with these anticipations. 


226 Thompson 


Group A, showing the effects of Excitation of the left Anterior Crural and 
Sciatic Nerves, the blood supply of the limb being unaltered. 


EE iit an heer (st ere iyie tw likilo. ll! 
Creatine Creatine “ 
Ex Waterd | tcueeee per cent. for| Total N Ash 
nash y D : water at per cent. per cent. 
(actual) = 
; 75 per cent. 
| 
l jR, 77021 “4917 5051 NSE 1:1881 
hs (as : 502 ee ; 
VL C415 4868 5025 1:1621 
2 R. 77°765 ‘5047 6224 35644 171206 
L 78°205 *5054 5269 3°4608 bee 
ap 75591 |  °5785 ‘5857 3°752 1-226 
L. 74°460 5974 “6010 3°822 1a 
ee 75°190° ‘5865 ‘5880 3°752 1-196 
L. 75°328 “5961 5987 3°794 1°237 
I 


In the table, two columns are given, the first showing the actual per- 
centage of creatine in the moist muscle on the two sides, the resting and 
the excited. The second column gives the percentages when the water is 
reduced to the same proportion, namely, 75 per cent. on both sides. It is 
only from these latter figures that any safe conclusion can be drawn con- 
cerning the effect of the excitation. On examining the figures it will be 
seen that there is either practically no change in the quantity of creatine 
in the muscles of the two sides, or that the difference is so small as to 
lie within the errors of the method. The greatest difference is shown 
in Experiment 3, where the creatine of the left side is ‘0153 per cent. 
higher than on the right—that is to say, an increase of 24 per cent., 
which cannot be said to be outside the limits of error. 

The conclusion, then, is that with intact circulation prolonged activity 
does not alter the proportion of creatine in voluntary muscles. It will be 
seen, also, that the effect on the content of water is negligible. 

Control experiments, in which all the operative procedures (including 
division of the nerves on the left side) were the same but without 
excitation, bear out this conclusion. 


ContROL ExperRIMENTs To GRoup A, in which the Left Anterior Crural and Sciatic Nerves 
were divided but not excited, the blood supply of the limb being unaltered. 


| Oreste Creatine : 
Exp. Water. per cent. | Pe cent. for; Total N Ash 
(actual) water at per cent. per cent. 
; 75 per cent, 


7 1°352 


75 
"755 1362 


75843 “6050 6117 3 
75°846 ‘6059 ‘6125 3 


iy Re 

tL 
6 R. 76°205 ‘5865 "5959 3°584 1:279 
L. 76°059 *5978 “6061 3°724 1°285 


The Metabolism of Voluntary Muscle 227 


A closer agreement in the analyses of the muscles of the two legs could 
not well be expected. Hence the operative procedures in themselves were 
without effect on the creatine content of the muscles. 

The second series of experiments includes those in which the main 
artery of the limb on the left side was ligatured immediately above 
Poupart’s ligament, before the excitation of the nerves was begun. The 
results obtained are given in the following table :— 


Group B, showing the effects of Excitation of the Left Anterior Crural and Sciatic Nerves 
after ligature of the External Iliac Artery. 


Creatine Total N 


Rep | Water beonenie per cent. for} Total N per cent. Ash 
gm ; ete ; water at per cent. Water per cent. 
ie acme by per cent. 75 per cent. 

wf R. 75293. | ‘4672 | -4690 | 3814 3873 

ae? 78312 | ‘4296 | 4486 | 3:556 3°764 
2 ( R. | 75°016 4899 | -4809 | 3-872 3°829 1-953 
L. 77°491 ‘4349 «| -4512 | 3-643 3°713 1-923 

g { R. 75°225 ‘4728 "4742 3 799 3°810 

L. 77°745 ‘4280 | 4436 3°416 3°541 
10 f B- 75-034 ‘4464 | +4446 rn wy 1215 
L. 74-239 ‘4215 | "A172 id 324 1°363 


In each of the four experiments there has been a loss of creatine in the 
excited muscles, accompanied by a reduction in the total nitrogen. There 
has also been in three of the four an increase of water. 

That these results are due to the stimulation and not to the operation 
is shown by a comparison with the following controls, in which all the 
conditions were the same except that excitation of the nerves was 
omitted :— 


Conrrot EXPERIMENTS TO Group B, in which the Left Anterior Crural and Sciatic 
Nerves were divided but not excited, the Left External Iliac Artery being ligatured. 


mene Creatine 
Ex Tipe a per cent.for| Total N Ash 
e ‘ actual) : water at per cent. per cent. 
; 75 per cent. 
ll R 76054 5698 ‘5779 4051 13614 
L | 75120 5834 5843 4-080 1°3812 
12 R 75963 5818 5893 3741 
L 76°215 5808 5902 3°717 
(R 75°832 4837 4891 3°570 


228 Thompson 


Taking the content of creatine from the column in which the water has 
been reduced to 75 per cent. on the two sides, the averages for the 
three experiments are as follows:—right leg, *5521 per cent.; left leg, 
‘5593 per cent. This gives a slight increase on the left side in the control 
experiments, but the alteration lies within the margin of error. The 
control experiments do not, at all events, show any decrease in the creatine 
of the muscles whose nerves have been divided. 

A fuller analysis of the results of the experiments in Group B brings 
out the following averages :— 


« | Loss per cent.| Loss per cent. 
ae leg ee in whole of each 
mesiine) i muscle. substance. 

Solids 24°822 22°151 2-671 10°76 
Creatine “A777 ‘4478 0299 6°26 
| Total nitrogen 3°837 3°673 164 4:26 


That is to say, as a result of prolonged activity with reduced blood supply 
there was a fall of 2°671 per cent. of the solids, of 0299 per cent. of 
the creatine, and of ‘164 per cent. of total nitrogen. These losses are 
equivalent respectively to 10°76 per cent. of the actual solids in the muscle, 
to 6:26 per cent. of the creatine, and to 4:26 per cent. of the total nitrogen. 
That the loss of total solids is greater than that of the creatine or total 
nitrogen indicates that non-nitrogenous substances are used up more freely 
under the conditions of excitation than creatine or other nitrogenous com- 
pounds. There can be little doubt that the solids which disappear are 
mainly organic non-nitrogenous bodies. The available analyses of ash 
show very little change in the active muscle, and therefore support this 


inference. 


If these results be compared with those of Group A, in which the blood 
supply of the active muscles remained unaltered, it will be found that the 
averages are as follows :— 


Loss or gain | Loss or gain 
Right leg Left leg per cent. per cent. 
(resting). (active). in whole of each 
muscle. substance. 
Solids 23°751 23°398 "453 loss 1°91 loss 
Creatine 5503 ‘5573 ‘0073 gain 1:27 gain 
Total nitrogen 3°689 3692 No change | No change 


Thus the loss in total solids was under 2 per cent. of the actual amount 
present; no change occurred in the total nitrogen; while the alteration in 


creatine shows a gain of 1:27 per cent. of the total present. 


tions are all within the limits of error of the methods. 


These varia- 


The Metabolism of Voluntary Muscle 229 


It may be concluded, therefore, that when the blood supply is adequate 
and the period of rest sufficient as compared with that of activity, there 
is, pari passu, a complete restitution of the substances used up by muscle 
in contraction. 

When, however, the blood supply is restricted, this does not occur. 
There is then a reduction of the store of energy-yielding solids held in 
the muscle, and the loss is not confined to non-nitrogenous organic sub- 
stances, but extends to the creatine as well. In what form the creatine 
leaves the muscle has not been ascertained. This and other problems 
concerned with the metabolism of active muscle have yet to be 
investigated. But it may be recalled that Weber (24), as also Howell and 
Duke (25), found creatine to be given off by the surviving heart to the 
fluid circulating through it. 

Analyses of the creatinine in the resting and active muscles by Folin’s 
recent method gave very discordant results, and no trustworthy deductions 
can be drawn from them. 


CONCLUSIONS. 


1. In voluntary muscle with intact circulation, excited intermittently 
for over two hours in periods of one-minute excitation followed by two- 
minutes rest, there is no loss of creatine, of total nitrogen, or of 
total solids. 

2. On the other hand, in voluntary muscle with restricted circulation, 
excited in the same way and for the same length of time, there is a loss 
of the solids amounting to over 10 per cent.; of creatine to over 6 per cent.; 
and of total nitrogen to over 4 per cent. 

3. It would appear, therefore, that under normal conditions with intact 
blood supply the substances used up to furnish the energy of contracting 
muscle are restored as fast as they are consumed. but if the blood supply 
be deficient the store of energy-yielding material is depleted, and the loss 
is not confined to non-nitrogenous material, but extends to creatine and 
possibly to other nitrogenous substances as well. 


LITERATURE REFERENCES. 


(1) Liesre, Annalen, 1847, Ixii. 257-369. 
: (2) Sarakow, ‘‘ Beitrag z. Physiologie d. Muskelstoffwechsels,” Virch. Archiv, 
1863, xxvill. 544. 

(3) Sczetkow, “Ueber Kreatingehalt d. Muskeln,” Centralbl. f. d. med. 
Wissensch., 1866, No. 31, p. 481. 

(4) Monarr, Atti R. Accad. d. Sci. d. ee 1887, xxii. 846-864; Abstr. 
Maly’s Jahresb., 1887, xvii. 311-312. 

(5) Nawnockt, * Beitrage z. teecliaciidsl im Muskel,” Centralbl. f. med. 
Wissensch., 1865, p. 417. 


230 The Metabolism of Voluntary Muscle 


(6) Vorr, C., ‘‘Das Verhalten d. Kreatins, d. Kreatinins u. d. Harnstofis im 
Tierkérper,” Zeitschr. f. Biol., 1868, iv. 77-98. 

(7) Meuuansy, E., ‘‘Creatine and Creatinine,” Journ. Physiol, 1908, xxxvi. 
447-487. 

(8) von Furrn, O., and Scuwarz, “Uber d. Verteilung d. Extraktivstoffes im 
Saugetier-muskel,” Biochem. Zeitschr., 1911, xxx. 413-432. 

(9) Scarripi, V., “Uber d. Verhalten d. Muskelkreatins b. d. Ermiidung,” 
Biochem. Zeitschr., 1913, 1. 402—417. 

(10) Granam Brown, T., and Carucart, E. P., “The effect of work on the 
creatine content of muscle,” Biochem. Journ., 1909, iv. 420-426. 

(11) PexeLHarine and Hoocennuyze, “Die Bildung d. Kreatins im Muskel b. 
Tonus u. b. d. Starre,” Zeitsch. f. physiol. Chem., 1910, lxiv. 262. 

(12) K. B. Hormann, “Ueber Kreatinin im norm. u. path. Harne,” Virchow’s 
Archiv, 1869, xlviii. 358. 

(13) P. Grocco, ‘‘ La creatinina in urine normali e patologiche,” Annal di chim, e 
di farmac., iv. s. 4, 211. (Quoted from Maly’s Jahresbericht, 1887, xvi. p. 199.) 

(14) Gregor, A., ‘Beitr. z. Physiol. d. Kreatinins,” Zeitschr. f. physiol. Chem., 
1900, xxxi. 98-118. 

(14a) J. Morressrgr, “Influence du travail musculaire s. |’élimination de la 
créatinine,” C.R. Soc. Biol., ili., series 9, 1891, p. 573. 

(15) HoogEnnuyze and VERPLOEGH, ‘“‘ Beobachtungen wu. d. Kreatininaussch. b., 
Menschen,” Zeitschr. f. physiol. Chem., 1905, xlvi. 415-471: ibid. 1908, Ivii. 
161-266. Also “Uber d. Einfluss v. Sauerstoffarmut auf d. Kreatininauscheidung,” 
ibid., 1910, lix. 101. 

(16) PEKELHARING, ‘Die Kreatininausch. b. Mensch. unter d. Einfl. v. Muskel- 
tonus,” Zeitsch. f. physiol. Chem., 1911, lxxv. 207. 

(17) Demant, B., “ Beitr. z. Kenntn. d. Extraktivstoffe d. Muskeln,” Zeitschr. f. 
physiol. Chem , 1879, iii, 381-390. 

(18) Menpex, L. B., and Ross, W. C., “Inanition and the creatine content of 
muscle,” Journ. of Bicl. Chem., 1911, x. 255-264. 

(19) Ranke, J., ‘Tetanus: eine physiol. Studie,” Leipzig, 1865. 

(20) Danitewskt, ‘ Ueber d. Ursprung d. Muskelkraft,” Charkow, 1846. 

(21) Worrz, E., “Ein Beitr. z. Chem. d. roten u. weissen Muskeln”’: Inaug. 
Dissert., Tiibingen, 1900. 

(22) Ganiks, M. E. A., “Contrib. a l’étude d. muscles en repos et en travail chez 
la grenouille,” Arch. d. sci. biol. d. St Pétersb., 1902-3, ix. 279. 

(23) Barcrorr and Karo, “Effect of functional activity upon the metabolism, 
blood-flow, and exudation in organs,” Proc. Roy. Soc. Lond., 1915, Ixxxviii. 541. 

(24) Weser, S., ‘Physiologisches zur Kreatininfrage,’ Arch. f. exp. Path. u. 
Pharm., 1908, lviii. 93-112. 

(25) Hows and Dukg, “ Note upon the effect of stimulation of the accelerator 
nerve upon the calcium, potassium, and nitrogen metabolism of the isolated heart,” 
Amer. Journ. Physiol., 1909, xxiii. pp. 174-179. 


THE ACTION OF THYROID UPON THE GROWTH OF THE 
BODY AND ORGANS OF THE WHITE RAT. By P. T. 
HERRING. (From the Physiology Department of the University of 
St Andrews. ) 

(Received for publication 6th March 1917.) 


CONTENTS. 


PAGE 
I. INTRODUCTION . 2 : : : : - PBA 
II. MerHops AND ventas Burioess , i ! ; Z 233 
Ill. Resutts OsraInEeD : ; : : 235 
(a) Effects of Thyroid on the Grow th of the Body . ; p : 235 
(b) Influence of Thyroid upon Individual Organs . ; ; , 236 “ 
1. The suprarenals ; : ; . : : 236 
2. The heart. : : : J : ‘ : 237 
3. The kidneys 3 ; F ; : 7 238 
4. The pancreas , : “ ; ‘ 239 
5. The liver ; . ‘ : , 3 J : 240 
6. The spleen : : z : ; : 241 
7. The thymus. : : : . P : ; 241 
8. Thetestes . j : ‘ : . ‘ : 242 
9. The ovaries . : ‘ ; ; : F 242 
10. The uterus . ; ? : ; ‘ : ; 243 
11. The thyroids : : : , : 243 
12, The pituitary body . ; ; : 244 
IV. Discussion or RESULTS : ; : : : , ; 245 
V. SUMMARY OF CONCLUSIONS F : : : ; F ; 248 
VI. LirerRAtTURE CITED ‘ : 3 B : : ; ; 249 
MIL. TABLES .. ; ; y i : 2 : z 250 


I. INTRODUCTION. 


MANny observations are now on record of the influence thyroid-feeding 
exerts upon the growth of the body. The earlier literature on the subject 
is discussed by Biedl (1) and Swale Vincent (20). References to the 
more recent literature are given in a paper by E. R. Hoskins (11). 
Comparatively few observations have been recorded of the action of 
thyroid upon individual organs. R. G. Hoskins (12) in 1910 described 
hypertrophy of the suprarenals in young guinea-pigs fed for fifteen days 
on small quantities of desiccated thyroid. Bircher (2) in the same year, 
in an experimental research upon the production of “Kropfherz,” found 
that rats, given water from certain wells alleged to produce the disease, 
showed diffuse colloid hypertrophy of the thyroids often accompanied by 
degenerative changes; there was also considerable hypertrophy of the heart. 
He ascribed these changes to the direct action of some toxic substance, and 


232 Herring 


considered that the condition was really associated with a deficiency of 
thyroid secretion although the thyroids were enlarged. Bircher also 
described acceleration of the development of the bony skeleton. 

Utterstrém (21) in 1910 fed rabbits with thyroid and described a 
resulting delay in the involution of the thymus. 

Iscovesco (13) in 1913 claimed to have isolated from thyroid tissue an 
ether-soluble material which, injected repeatedly into rabbits, produced 
remarkable effects. Iscovesco noted hypertrophy of the suprarenals in 
both sexes, of the ovaries and uterus in the female, and of the testes in 
the male. The spleen was slightly increased in size, the heart more 
enlarged in males than in females, the kidneys hypertrophied in males 
only. The growth of the body as a whole was greatly accelerated in 
young rabbits, but only up to a certain age, after which administration of 
the ether-soluble material caused actual loss of weight. In this connexion 
the results obtained by Gudernatsch (6) are interesting. Gudernatsch 
found that the macroscopic changes of differentiation in the tadpole are 
greatly accelerated by even very small amounts of thyroid administered as 
food. Growth is, however, checked, and the tadpoles rarely survive the 
accelerated metamorphosis thus produced. 

Livingston (17) in 1914 showed that the hypertrophy of the pituitary 
body which is known to follow thyroidectomy in rabbits is prevented by 
feeding the animals with thyroid subsequent to operation. 

Kojima (16) in 1916 described hypertrophy of the pancreas, the 
occurrence of numerous mitotic figures in the nuclei, and alterations in 
the granules of the cells of the pancreas as the result of thyroid-feeding. 

The author (9) (10) in 1916 confirmed the results obtained by E. R. 
Hoskins on the suprarenals, and found that small doses of thyroid pro- 
duce hypertrophy of the suprarenals and an increase in their adrenalin 
content both in the cat and in the white rat. Hypertrophy of the heart 
was noted in the rat, and in a communication given to the physiological 
section of the British Association at Newcastle in September 1916 figures 
were shown which demonstrated that many other organs are affected, the 
suprarenals, heart, and kidneys being especially subject to hypertrophy as 
the result of thyroid-feeding. Further observations were in progress when 
a very complete paper on the subject appeared by E. R. Hoskins (11). 
Hoskins fed white rats with small doses of thyroid, thymus, pituitary, 
and pineal body. The chief changes occurred in the thyroid-fed groups. 
Compared with normal control animals of similar sex and of the same 
litter, the thyroid-fed rats showed hypertrophy of the suprarenals, heart, 
kidneys, alimentary canal including pancreas and mesentery, liver, and 
spleen. In males the testes were somewhat increased in size, and in 
females the ovaries. The pineal body showed no appreciable changes, the 
thyroids were rather smaller, and there was some delay in growth of the 
pituitary body in females. There was no constant change in the thymus. 
Hoskins also investigated the head, body, cartilaginous skeleton, integu- 


The Action of Thyroid upon the Organs of the White Rat 233 


ment, brain, and lungs, and noted a small increase in the weight of most 
of these in the thyroid-fed rats. His animals were observed over long 
periods, but were given very little thyroid. The gross body-weights of 
the thyroid-fed animals averaged very little more than those of the control 
animals, so little, indeed, that Hoskins doubts if the increase of weight is 
of any special significance. The thyroid-fed animals lose an amount of 
fat which nearly counterbalances the increase of weight of other structures. 
Hoskins points out, that, taking this into consideration, his results are not 
incompatible with the view expressed by Moussu (18) and Schafer (19), 
that small doses of thyroid produce an increase in body weight. On the 
other hand, Hoskins agrees with a large number of observers cited in his 
paper that larger does of thyroid bring about a decrease or retardation of 
body weight. 

My own results embodied in this paper are in general agreement with 
those of Hoskins. I have employed somewhat larger doses, or doses of 
similar size more frequently administered, and it is interesting to note that 
the degree of hypertrophy obtained in several organs is much greater than 
that obtained by Hoskins. In other organs my figures more nearly 
approach those of Hoskins, and may be of service in confirming and 
emphasising his results. Hoskins ascribes the hypertrophy to the 
increased rate of metabolism promoted by thyroid-feeding, and compares 
his results to a somewhat similar condition described by Hatai (7) in 
which hypertrophy of organs is induced by increased exercise. The degree 
of hypertrophy reached by some of the organs in my series seems to me to 
point to something further than this, and suggests a specific influence of 
the thyroid upon certain structures of the body. 


II. MerHops AND MATERIAL EMPLOYED. 


The experiments have been carried out on white rats exclusively. The 
male animals in the first series were young adults born in the spring of 
1916, and were mostly from stock reared for several generations in this 
laboratory. A few were from an outside stock, but the controls and thyroid- 
fed animals were in this case paired against one another. No essential 
differences were noticed in the control animals of the two stocks. The 
female rats were all from the laboratory stock, and each thyroid-fed 
animal and its control were taken from the same litter. These animals 
were born in the autumn of 1916. According to Jackson (14) and King 
(15) the variability in body weight of albino rats of the same litter is only 
about half as great as that due to general racial variation. Even in the 
same litter, however, the individuals of the same sex often show consider- 
able differences in weight and disposition, and for that reason animals as 
like as possible in the same litter were selected. 

The rats were isolated from one another during the period of experi- 
ment, and kept each in a separate cage. The isolation in some cases 


934 Herring 


appeared to affect growth, and is therefore not altogether desirable. 
Hoskins avoided this objection but was able to do so in that he adminis- 
tered thyroid at longer intervals. My rats took the greater part of their 
food at night, and to ensure that each animal received its full dose of 
thyroid the thyroid was finely minced and intimately mixed in the upper 
layers of bread and milk more than sufficient for the twenty-four hours. 
The thyroid was obtained fresh from the ox, and administered in daily 
amounts of from 0°1 to 0°2 grm. Some of the control animals received 
daily equivalent amounts of fresh ox flesh similarly minced and mixed 
with the bread and milk. 

The weights of the animals were recorded weekly, and finally at the 
end of the experiment. A thyroid-fed animal and its control were killed 
by chloroform and bled by section of the large veins in the thorax. The 
contents of the stomach, if any, were removed and weighed, and their 
weight deducted from the weight of the animal. The final weights of the 
animals recorded are therefore the net weights after deduction of the 
contents of the stomach. The organs investigated were dissected out, 
cleaned of fat and connective tissue, and weighed. In many cases the 
adrenalin contents of the suprarenals were also measured by Folin’s 
method. Pieces of the organs were preserved in Zenker’s fluid, cut and 
examined. <A complete account of the histological changes, however, is not 
undertaken in this paper. 

Details of the records obtained are given in Tables III. to VIII. At the 
foot of each table the average weights of the organs of the animals 
employed are given, then the average weights per 100 grm. body weight, 
and lastly the average weights per 100 mm. final body length. The body 
length is the distance from tip of nose to centre of anus in the extended 
animal after death. 

The organs investigated are the suprarenals, heart, kidneys, pancreas, 
liver, and spleen in both male and female rats; the thymus, thyroids, and 
pituitary body in females only. The weights of the testes in males, and of 
the ovaries and uterus in females, are also given. In Tables J. and II. the 
average weights of the different organs in male and female of both the 
control and the thyroid-fed groups are recorded for convenience of com- 
parison in terms of average weights per 100 grm. body weight and per 
100 mm. body length. In the animals in which the administration of 
thyroid has notably interfered with the normal gain of weight of the body 
as a whole, especially by the loss of fat, a comparison of the average 
weights of the organs per unit body length probably gives one a more 
exact figure of the changes which have taken place than does a comparison 
in terms of body ‘weight. Both methods are adopted in this paper. 

Individual differences in the relation between body length and body 
weight are considerable in the rats examined in this laboratory. Com- 
pared with Donaldson’s tables (4), both the male and female rats show a 
greater body weight per unit length than those of the Wistar Institute. 


The Action of Thyroid upon the Organs of the White Rat 235 


The rats recorded in Table V. agree more closely with the measurements 
of the Wistar Institute rats. It is possible that my observations on the 
length of the animals have not been taken strictly in accordance with 
the routine method adopted in the Wistar Institute. Alterations in the 
position of the animal readily alter the observed length, and my measure- 
ments may have been taken with too little extension of the body and neck. 
The measurement shrinks a little after rigor mortis has set in, but my 
records were always taken immediately after death when the muscles are 
lax. As the same relative differences are noted in both the control and 
thyroid-fed animals, this does not alter their value for comparison with one 
another, but a comparison with the Wistar Institute rats is not so satis- 
factory. The weights of the individual organs compared with the body 
weight show a greater resemblance to the weights recorded in Donaldson’s 
tables. Differences are noted later in the description of changes in the 
individual organs resulting from thyroid-feeding. It is possible, of course, 
that the rats bred in this part of the world have their own characteristic 
features. There are certainly peculiarities in individual organs, such as 
the spleen and liver, which differ widely from those noted in the Wistar 
Institute. 


III. Resutts OBTarNep. 
(a) Effects of Thyroid on the Growth of the Body. 


Considerable variations in growth are apparent both in the control and 
thyroid-fed animals, male and female. Some of the controls received small 
doses of fresh ox flesh, but with no obvious effect upon the rate of their 
growth. The thyroid-fed male animals in Table IV. increased in weight 
fairly uniformly, but as the corresponding initial weights of the control 
animals were not recorded, no definite comparison can be made between 
them. It can only be said that the thyroid did not perceptibly increase or 
check the rate of growth. In Table V. a comparison can be made; the 
control rats gained an average of 69 grm. in twenty-five days, while the 
animals under the influence of a daily dose of 0:2 grm. thyroid gained an 
average of 65 grm. in the same time. The control animals, therefore, 
gained slightly more than the thyroid-fed animals. The same is true of 
the female rats, Tables VII. and VIII, in which the controls gained on the 
average 2 grm. more than the rats receiving 071 grm. fresh thyroid daily. 

The average among the females is made up of widely differing figures. 
Some rats appeared to thrive and put on weight under the influence of 
thyroid much better than their controls; in other animals a decided check 
was imposed on their growth by thyroid-feeding. In a few instances 
thyroid appeared to stimulate a more rapid growth which was later 
followed by an actual decline in weight, and some of the final figures of 
weights of the thyroid-fed animals are less than the weights attained at 
an earlier stage of the experiments. Some of the animals died while under 
the influence of thyroid; one of them, a female, lost weight from the first 


236 Herring 


exhibition of 0:1 grm. thyroid, its control steadily gaining. These two 
animals are not included in the returns. The fourth rat in Table VIII. 
gained 15 grm. in thirty days, while its control gained only 8 grm. On the 
other hand, the first rat in Table VII. gained 23 grm. in twenty-one days, 
its sister under thyroid only gaining 1 grm. in the same period. Several 
of the thyroid-fed rats died in their cages, one, a male, without any 
previous loss of weight. Another male lost weight for several days before 
death, and the females that died also lost weight, and several showed 
symptoms of diarrhcea. None of the control rats showed any adverse 
symptoms, and none died in their cages. 

The female rat appears to be more susceptible to the influence of 
thyroid than the male. In all the thyroid-fed animals there is loss of fat, 
sometimes to an extreme degree, and an absence of glycogen from the liver. 
This loss of weight may be counterbalanced in part or wholly by increase 
in the weight of other structures, The longest period over which thyroid 
was given in these experiments was fifty-one days, and the dose 0:2 grm. 
fresh thyroid daily. The animal, a male, throve well at first, but finally 
died. Its organs show an extreme degree of change (Rat No. 7, Table IV.). 
There was a complete absence of fat, but an enormous hypertrophy of 
heart, kidneys, and liver—a condition probably incompatible with a 
further prolongation of life. 

Hoskins, giving smaller doses, found that thyroid had little definite 
influence on the growth of the body asa whole. 

My figures support his conclusions. No comparisons can be made as 
regards alterations in length of body, as the initial lengths were not 
recorded. In the male animals the average final body lengths are rather 
greater in the thyroid-fed than in the controls. In the female rats the 
lengths of both groups coincide. It is unlikely, therefore, that there is any 
decided increase or check imposed on growth by thyroid-feeding. 

Daily doses of 0:1 to 0:2 grm. of fresh ox thyroid are undoubtedly toxic 
to white rats. They may not influence to any extent the growth of the 
body as a whole, but they bring about changes in the body which 
ultimately prove fatal. 


(b) Influence of Thyroid upon Individual Organs. 


1. The Suprarenals.—The suprarenal bodies are constantly increased 
in size by thyroid-feeding both in male and female rats. The increase is a 
rapid one, as is shown in Table VI., where the administration of 0:2 grm. 
thyroid daily to young male rats has brought about an increase of 73 per 
cent. in weight per 100 grm. body weight in eight days. 

Compared with Donaldson’s tables, the weights of the suprarenals of 
the male control animals average rather less per unit body weight than 
those of the Wistar Institute rats. The suprarenals of the female animals 
agree very closely with Donaldson’s figures. The highest weight recorded 
in the male thyroid-fed rats is 60 mg. in a rat weighing 148 grm. 


The Action of Thyroid upon the Organs of the White Rat 237 


According to Donaldson’s figures, the average weight of the suprarenals for 
an animal of that body weight is 28 mg. Among the controls of the same 
experiment a rat of the same weight has suprarenals weighing 25 mg. 
The increase of weight of the suprarenals is, therefore, 113 per cent. against 
Donaldson’s figure, and 140 per cent. against the control. 

In female rats the suprarenals are normally heavier per unit body 
weight than in males, and the percentage increase of weight caused by 
thyroid-feeding is less. The heaviest suprarenals found in the female rats 
weighed 74 mg. in an animal of 140 grm. body weight fed for thirty-six 
days on 0:1 grm. thyroid daily. Donaldson’s average for a female rat of 
140 grm. body weight is 38 mg. In this rat the suprarenals are 94 per 
cent. heavier than the normal average. 

The examples quoted, however, are the extremes. The general averages 
show an increase of 56 per cent. per 100 grm. body weight in the males, 
and an increase of 41 per cent. per 100 grm. body weight in the females 
as the result of thyroid-feeding. The increase in weight is partly of 
medulla but chiefly of the cortex. 

In male rats the author (10) has shown that thyroid-feeding increases 
the adrenalin content absolutely, but diminishes it relatively to the per- 
centage weight of the suprarenals. The increase of adrenalin in the supra- 
renals of the female rats resulting from thyroid-feeding is less constant. 
In the female rats which thrive well under the influence of thyroid the 
adrenalin is increased, but in those animals which show checked growth 
and other toxic symptoms the amount of adrenalin is actually decreased. 
Observations upon this subject are still being made, and will be dealt with 
in a subsequent paper. They may have an important bearing upon the 
explanation of the factors in thyroid-feeding which bring about the great 
hypertrophy of some of the organs of the body. 

My results agree very closely with those obtained by Hoskins; but 
the hypertrophy of the suprarenals is greater in my series than in his, 
probably because of the larger amounts of thyroid given. Hoskins noted 
that the reaction of the suprarenals to thyroid is relatively greater in 
males than in females, a view which my figures confirm. 

2. The Heart.—The heart is constantly and very greatly hyper- 
trophied under the influence of thyroid. The response is an immediate 
one. ‘Table VI. shows that the administration daily of 0-2 grm. fresh 
thyroid to young male rats increases the weight of the heart by 78 per cent. 
in eight days. Prolonged feeding with thyroid leads to a much greater 
increase. 

The weights of the hearts of the control male rats agree very nearly 
with the normals of Donaldson’s tables; the control females have rather 
heavier hearts than the females of corresponding body weight in the 
- Wistar Institute. The largest heart recorded in my series (No. 7, Table IV.) 
is in a thyroid-fed male rat, and weighs 2:455 grm., the body weight of 
the animal being 182 grm., and the body length 189 mm. For a male rat 


238 Herring 


of this weight Donaldson’s normal figure for the heart is about 0°745 grm. 
Among the control rats in Table III., No. 6, a larger animal weighing 
200 grm., and having a body length of 193 mm., has a heart which weighs 
0'754 grm. This figure corresponds very well with Donaldson’s normal, 
and is, indeed, slightly less per unit body weight. The size of the heart 
of the thyroid-fed animal is more than trebled, and its weight is 230 per 
cent. above the normal. This rat was fed on 0:2 grm. fresh ox thyroid 
daily for fifty-one days. Among the females the heaviest heart is also 
found in the animal which received the largest amount of thyroid. This 
rat (No. 7, Table VIII), with a body weight of 138 grm., and a length of 
174 mm., has a heart weighing 1°187 grm., and received 0:1 grm. of thyroid 
daily for forty-six days. The normal weight of the heart of a female rat 
of this size is about 0°600 grm., the gain of weight, therefore, in the thyroid- 
fed animal is about 98 per cent. These are the extremes, but the averages 
are also high. The increase in weight of the heart of the thyroid-fed 
males averages 123 per cent., and of the thyroid-fed females 59 per cent. 
If the weights of the heart per 100 mm. body length be compared, the 
figures are not much altered. The male thyroid-fed animals show an 
increase of 116 per cent., and the females an increase of 65 per cent. 

In both male and female rats the heaviest hearts are in the animals 
which received most thyroid. The female heart is normally heavier than 
the heart of the male of the same body weight. The females received less 
thyroid than the males, and their hearts are not so greatly hypertrophied. 
The degree of hypertrophy of the heart is much greater than that noted 
by Hoskins. He obtained a hypertrophy of 16:7 to 246 per cent. in 
females, and of 15-4 to 36 per cent. in males. As already stated, the doses 
of thyroid he used were smaller or given at less frequent intervals. 

The increase in size affects the whole heart, and the naked-eye appear- 
ance is very striking. The ventricles are large and muscular. On section 
the increased thickness of their walls is apparent, that of the left ventricle 
being particularly so. Microscopical examination shows an increase in 
muscular fibres, and an apparent increase in size of many of the fibres. 

3. The Kidneys.—The kidneys also are constantly and greatly hyper- 
trophied by thyroid-feeding both in male and female rats. 

The weights of the kidneys of the control animals agree fairly well 
with Donaldson's figures per unit body weight, but are somewhat heavier in 
the males of my series and lighter in the females. The greatest changes 
are again noticed in the rats which received most thyroid. The last male 
rat in Table IV. shows a weight of 4°758 grm. for both kidneys. This rat 
has a body weight of 182 grm. and a length of 189 mm. Donaldson’s 
average weight for the kidneys of an animal of this weight is about 
1574 grm. Among the control animals of Table IIL, No. 6, a larger rat, 
has a kidney weight of 1388 grm. The weight of the kidneys of the 
thyroid-fed rat is more than trebled, the increase against Donaldson's 
figure being 201 per cent., and against the control 214 per cent. 


The Action of Thyroid upon the Organs of the White Rat 2389 


Among the female rats No. 6, Table VIII, shows kidneys weighing 
2:036 grm. This rat received 0:1 grm. fresh thyroid for thirty-six days, 
and has a body weight of 140 grm., and a length of 174mm. Donaldson’s 
average for the kidneys of a female rat of this size is about 1°260 grm. 
The kidneys of the thyroid-fed animal show an increase of weight amount- 
ing to 61 per cent. 

The average increase in weight of the kidneys of the thyroid-fed 
animals per 100 grm. body weight over that of the controls amounts to 
106 per cent. in the males, and 63 per cent. in the females. Per 100 mm. 
body length the increase is 100 per cent. in the males, and 70 per cent. in 
the females. 

The kidneys of the normal rat are relatively heavier in the female than 
in the male. The increased weight due to thyroid feeding is greater in 
the male than in the female, but the males received more thyroid. The 
increase of weight seems to be roughly proportional to the amount of 
thyroid given. 

Hoskins with smaller doses obtained an increase in weight of the 
kidneys varying from 33 to 46:3 per cent. in females, and from 40-4 to 
44°4 per cent. in males. 

Histologically the enlarged kidneys of thyroid-fed animals show an 
increase in the size of the tubules and a looser texture of the kidney 
generally. In the extreme enlargements there is a decided increase in 
the amount of connective tissue in the kidney. 

4. The Pancreas.—No figures relating to the weight of the normal 
rat's pancreas are given in Donaldson’s memoir. Hoskins describes an 
increase in the weight of the alimentary canal, including pancreas and 
mesentery, as the result of thyroid-feeding. Kojima (16) found evidences 
of greatly stimulated growth in the abundance of karyokinetic figures in 
the pancreas induced by thyroid-feeding. 

The measurements of the weights of the pancreas given in this paper 
are probably not quite correct. It is difficult to distinguish some of the 
rats’ pancreas from fat, and being a diffusely spread organ it is mixed with 
a good deal of fat and connective tissue. The pancreas was carefully and 
widely removed, spread out on a dark surface, and as much fat, lymph 
glands, and mesentery removed as possible. This is less difficult in the 
thyroid-fed animals, in which there is little or no fat. The weights in both 
groups of animals are probably in excess of the true value, but rather more 
so in the normal animals than in the thyroid-fed. 

Thyroid-feeding gives rise to a large hypertrophy of the pancreas in 
both sexes. The average figures show an increase in weight of 111 per 
cent. in the male thyroid-fed animals, and an increase of 80 per cent. in 
the female thyroid-fed rats. Per 100 mm. body length the increase is 
104 per cent. in the males, and 87 per cent in the females. 

According to the figures recorded, the weight of the pancreas per 


100 grm. body weight is rather greater in females than males, but per 
VOL. XI., NO. 3.—1917. 16 


240 Herring 


100 mm. body length the male pancreas is rather heavier than the female. 
For the reasons already stated it is doubtful how far these figures may be 
regarded as correct. There is sufficient evidence, however, to support 
Kojima’s work, and to substantiate the fact that thyroid-feeding brings 
about a great hypertrophy of the pancreas in both sexes. 

Histologically the increase of size of the pancreas appears to be due 
to an increase in the size and number of the alveoli. It is difficult to 
determine whether there is an increase in the islets of Langerhans. In 
the pancreas of the thyroid-fed cat the islets of Langerhans are more 
prominent than usual, and I have long been in the habit of giving prepara- 
tions of pancreas from thyroid-fed animals to the histology class, because of 
this prominence of the islets. 

5. The Liver.—The weight of the liver shows great variations in my 
series of rats, but in all the control animals, male and female, the liver 
weighs considerably less than that of the Wistar rat of the same body 
weight or body length. The animals reared in this laboratory have much 
lighter livers than the animals on which Donaldson’s tables are based. 
In my series, too, the liver is relatively heavier in the female than it is 
in the male, a feature also shown in the Wistar Institute rats, but less 
pronounced in them than it is in the rats reared in this laboratory. In 
Hoskins’ series the male liver is relatively larger than the female liver, 
but both are lighter, except in young males, than Donaldson’s figures. 

Thyroid-feeding increases the weight of the liver, but not to such an 
extent as is found in the organs already detailed. The liver of the male 
animals shows an average increase of 45 per cent. per unit body weight, 
and the liver of the females an average increase of 81 per cent. Measured 
per 100 mm. body length, the increase is 40 per cent. in the males and 
36 per cent. in the females. The increase in the males is largely helped 
by the last rat of the series, which has an enormous liver. The averages 
of the liver weights in the thyroid-fed animals, male and female, 
approximate very nearly to the figures given by Donaldson as normal. 
Hoskins obtained an enlargement of 64 to 24°4 per cent. of the livers 
of male rats, and of 267 to 30°5 per cent. of the livers of female rats, 
as a result of thyroid-feeding. My figures denote a larger increase in 
the male, but approach Hoskins’ figures for the percentage increase in 
the female. The hypertrophy is only excessive in the case of the one 
rat mentioned, No. 7 of Table IV., a rat which received the most thyroid. 

The enlargement of the liver found in the thyroid-fed animals is 
probably not incompatible with Hoskins’ explanation that it is due to. 
increased metabolism. The animals fed on thyroid ate more food than 
the controls, and in some of them the increased food intake was very 
pronounced. In all the thyroid-fed animals in which a special examina- 
tion was made there was an absence of glycogen from the liver, whereas 
in the control animals glycogen was abundant. Cramer and Krause (3) 
described the disappearance of glycogen from the livers of thyroid-fed 


The Action of Thyroid upon the Organs of the White Rat 241 


animals, and attributed it to an inhibition of the glycogenic function of 
the liver and not to an increased utilisation of carbohydrates. In light 
of the present observations the disappearance of glycogen may perhaps 
be ascribed simply to the increased rate of metabolism induced by thyroid. 
It is further possible that the increased production of adrenalin, already 
described in the suprarenals, may also exercise its influence on carbo- 
hydrate metabolism. 

6. The Spleen.—The spleen is another organ which shows great 
variations in size in the white rat. Donaldson includes in his series 
the weights of small spleens only, and rejects data from animals with 
so-called “enlarged spleens.” In my animals the small spleen is rarely 
met with; the usual spleen is nearly twice as large as that of the Wistar 
Institute rat. Hoskins noted the same difference in his rats, and questions 
the reliability of Donaldson’s tables for the weight of the normal rat’s 
spleen. Jackson (14) finds that the weight of the spleen varies directly 
with that of the liver, and looks for a close correlation between the two 
organs. My rats show a certain tendency to conform to this rule but 
by no means constantly. In neither male nor female control rats is the 
heaviest liver accompanied by the heaviest spleen, or the lightest liver 
by the lighest spleen. On the contrary, among the male control animals 
the lightest liver is associated with the second heaviest spleen, and the 
same is the case among the female rats. 

Thyroid-feeding as a general rule increases the weight of the spleen, 
but not to any great extent. Per 100 grm. body weight the average 
increase in weight of the spleen in the thyroid-fed animals is 39 per cent., 
that of the females 22 per cent. Per body length the increase is 35 per 
cent. in males, and 27 per cent in females. Considerable irregularities 
occur in the tables, and the increase in weight of the spleen of the thyroid- 
fed animals is not a constant phenomenon. The lightest spleen recorded in 
the series is in a thyroid-fed female, No. 2, Table VIII.; this, of course, may 
be one of the small spleens, but the control animal from the same litter has 
a much larger spleen. This animal also affords an exception to the general 
rule that the female possesses a relatively heavier spleen than the male. 
Further, the small spleen in this animal is associated with a large liver. 

Hoskins obtained an increase in weight of the spleen in his thyroid-fed 
rats, and also noticed great irregularity in the amount of enlargement. My 
results are sufficiently like his to confirm the opinion that thyroid-feeding 
does enlarge the spleen. The enlargement, however, is not such as to point 
to any special action of thyroid upon it, but rather to a general effect 
resulting from the increased rate of metabolism induced by thyroid-feeding. 

7. The Thymus.—The weight of the thymus was only measured in 
the female series of rats. In the male animals, which were rather older 
as a group than the females, the thymus was small and almost impossible 
to separate accurately from fat and connective tissue. The impression 
reached by mere examination was that thyroid-feeding hastens the involu- 


949 Herring 


tion of the male thymus. A few measurements made in another series of 
male animals support this view, but the difference is too small to be of any 
special significance. 

In the females the figures show an average rise of 7 to 11 per cent. in 
the weight of the thymus of the thyroid-fed animals. The delayed involu- 
tion in the female rat is not constant, and the figures are quite capable of 
explanation by normal variability. 

Hoskins found that the thymus of his thyroid-fed female rats averaged 
10 per cent. lighter than the thymus of the controls. In males there was 
no definite change. Hoskins concludes that thyroid-feeding has no 
constant effects upon the thymus, and my figures also support this 
conclusion. 

8. The Testes.— The testes show variations in weight, but not to 
anything like the extent characteristic of other organs, such as the liver, 
spleen, and thymus. No data are available for comparison in Donaldson's 
tables, because in his figures the epididymis is included with the testis. 
The epididymis shows large variations, and there is no advantage gained 
by including it with the testis as a single unit. 

The thyroid-fed animals in my series show slightly heavier testes than 
the controls, the increase being 8 per cent. per unit body weight, and 
5 per cent. per unit body length. Hoskins also obtained an increase of 
weight of the testes of his thyroid-fed animals, the enlargement amounting 
to 7 per cent. in the older animals, and 13 per cent. in the younger. In 
my series of control rats the first two animals have exceptionally small 
testes, and if they are excluded there is little difference between the 
averages of the remaining animals of the two groups. Hoskins attached 
no special significance to the increase of weight. The fact that our figures 
vary in the same direction may point to a slight influence of thyroid in 
accelerating the growth of the testes. Iscovesco also noted the same 
action. The acceleration is only small, and lies within the normal limits 
of variation. It is unlikely, therefore, that it can be ascribed to any direct 
action of thyroid, and is merely the result of increased metabolism. In 
my series it is associated with a somewhat hastened involution of the 
thymus, but in Hoskins’ rats this is not the case. 

9. The Ovaries.—The ovaries are liable to extreme variations in size, 
and a systematic examination of a larger number of animals than is included 
in this series is necessary to afford any sound conclusions as to the signifi- 
cance of changes brought about by thyroid-feeding. The ovaries of the 
thyroid-fed animals average 12 to 14 per cent. heavier than those of the 
control rats. This increase, it will be noticed, is associated in these animals 
with a seemingly delayed involution of the thymus. 

Hoskins obtained no constant results in his thyroid-fed animals. 
Iscovesco reported hypertrophy as the result of his ether-soluble extract 
of thyroid. 


The figures given in my records do not indicate that thyroid has any 


The Action of Thyroid upon the Organs of the White Rat 2438 


special action on the ovaries, but it is probable that their growth is to 
some extent accelerated by the general change in the body produced by 
hyperthyroidism. 

Histologically no essential alteration is found in the ovaries of the 
thyroid-fed animals. 

10. The Uterus.—The uterus in my measurements includes the two 
cornua and the Fallopian tubes to their attachment to the ovaries. The 
lower end was separated by a transverse cut through the junction of uterus 
and vagina. The line of junction is not very definite, but can be felt by 
compressing it between finger and thumb. Slight deviations from the 
junction do not materially affect the observed weight of the organ. No 
data are available for comparison with my figures as far as I am aware. 
The uterus shows extreme variations in weight. In none of the measure- 
ments was there any sign of the uterus being in a period of cestrus at the 
time. All the animals were virgins. 

In the thyroid-fed rats the general impression formed was that the 
uterus is distinctly retarded in development. A striking exception is 
furnished by No. 6 rat. In this animal, which received 0:1 grm. of thyroid 
for thirty-six days, the uterus is exceptionally large, weighing more than 
those of five of the other animals added together. Even with the inclusion 
of this organ the uteri of the thyroid-fed animals average 11 to 14 per cent. 
less by weight than those of the control rats. With the exclusion of No. 6 
rat and its control the weights of the uteri of the remaining six animals 
average 162 per cent. less than those of the controls. 

Iscovesco noted a large hypertrophy of the uterus in rabbits treated 
with thyroid material. There is no evidence in my figures of this being 
the case in rats except in the one instance mentioned. This animal was 
the largest of the series, and had a large pituitary body and a moderately 
large thymus. 

It is interesting to note that the small uterus is associated in my 
series with a small pituitary body, and to some extent with a relatively 
large thymus. There are exceptions, however, particularly with regard 
to the thymus, and my figures do not bear out any satisfactory establish- 
ment of a correlation between thymus and uterus. The correlation between 
uterus and pituitary body is more striking. The normal variability of 
the uterus is so great that these figures may have no special significance. 
On the other hand, it is possible that in young animals thyroid may restrain 
the development of the uterus indirectly by its action on the pituitary body. 
Further observations are necessary on this subject. 

11. The Thyroids.—Measurements of the thyroids were only made 
in the female animals of the series, but several have been made in males 
since the tables given were drawn up. The thyroids of both males and 
females were examined histologically. 

The normal weight of the thyroids is liable to considerable variations. 
The general average in the control animals is less than that of the Wistar 


24.4 Herring 


Institute rat. Hoskins also found a lighter series of thyroids in his 
animals, and ascribes the difference to differences of technique employed 
in their removal. In my series the thyroids were carefully dissected out, 
examined with a strong lens, and the outlying fat and connective tissue 
removed. 

Thyroid-feeding gives a reduction in weight of the thyroids in my series 
of 21 to 24 per cent. A similar but rather larger loss of weight occurs in 
the thyroids of the thyroid-fed males examined. The loss of weight is a 
very constant phenomenon, and I have observed the same thing in the 
thyroids of thyroid-fed cats. Histological changes also are constant and 
striking. The thyroids of the thyroid-fed animals present vesicles con- 
taining a denser and more stainable colloid, but the chief difference lies in 
the cellular elements of the thyroids. In the thyroid-fed animals the cells 
are thin and flattened, whereas in the control animals the cells are more 
numerous and cubical. The flattened cell appears to be associated with 
decreased activity of the gland, the cubical cell and comparatively little 
colloid with the more active type of gland. 

Hoskins noted no constant changes, though his figures, resulting from 
much smaller doses of thyroid, do not contradict mine. From the evidence 
adduced it seems clear that thyroid-feeding inhibits to some extent the 
action of the animals’ own thyroids, and in growing animals checks the 
simultaneous growth of these glands. Iscovesco found enlargement of 
the thyroids, but he used ether-soluble extracts, and his experiments do 
not necessarily afford a fair comparison with those in which thyroid itself 
is used, 

12. The Pituitary Body.—The weights of the pituitary body of the 
female control rats recorded in the tables agree fairly well with Donaldson’s 
figures, but average rather less per unit body weight. The pituitaries of 
the male animals were not recorded but were preserved for histological 
examination attached to the base of the brain. A few additional observa- 
tions have been made on male animals, but are not as satisfactory as they 
might be owing to the great loss of body weight which occurred in the 
thyroid-fed animals of the same litter. The pituitaries of the male controls 
are rather lighter per unit body weight than those of the females. This 
difference between the male and female pituitaries was noted by Hatai (8), 
and is shown in Donaldson’s tables. It is also seen in Hoskins’ figures. 

Hoskins found that the administration of small amounts of thyroid 
checks the growth of the pituitary body in the female rat, and accelerates 
it in the male. He obtained a decrease of weight of 8°8 to 11°5 per cent. in 
females, and an increase of weight of 18 to 21:3 per cent. in males. My 
figures confirm Hoskins’ results with regard to the females, and partly, 
but not entirely, with regard to the males. In the female thyroid-fed 
rats my results are almost identical, the pituitary showing an average 
decrease in weight of 11 per cent. when measured per 100 grm. body 
weight, and of 8 per cent. when measured in terms of body length 


The Action of Thyroid upon the Organs of the White Rat 245 


In the males examined the increase of weight of the pituitary of the 
thyroid-fed animals is 18 per cent. per 100 grm. body weight, but measured 
in terms of body length the thyroid-fed pituitary shows a decrease of 
18 per cent. compared with that of the controls. The comparison in terms 
of body length is probably more accurate where much weight has been lost 
during the period of thyroid-feeding, as was the case in these animals. The 
experiments are not numerous enough to settle this matter, but as far as 
they go they tend to show that thyroid-feeding also checks the growth of 
the male pituitary. 

The diminished size of the pituitary body seems to be chiefly scéoaated 
for by a delay in growth of the pars glandularis. There is considerable 
change in this lobe, and a notable deficiency in the number of the 
characteristic large granular cells. 


IV. Discussion OF RESULTS. 


It is generally agreed that the thyroid glands contain some substance 
or substances which, when liberated into the body, promote metabolism. 
A condition of hyperthyroidism may be set up by excessive activity of 
the thyroid glands themselves or by the administration of thyroid in the 
food. In the latter case the thyroid glands of the animal appear to 
take little or no part in the production of the hyperthyroidism. They 
are smaller than those of the control animals and show signs of relative 
inactivity. 

A small degree of hyperthyroidism may, in a young animal, to some 
extent accelerate the growth of the body, but in doses of 0:1 to 02 grm. 
fresh thyroid given daily tends rather to check the increase of weight 
in growing rats. Individual variations occur, but I am inclined to agree 
with Hoskins in this respect. Little or no alteration is produced in the 
body length, but there is, on the whole, a diminution of body weight. 
Thyroid-feeding, moreover, gives rise to profound changes in the body, 
and if long continued in the amounts mentioned brings about fatal results. 

Thyroid acts upon the metabolism of the body as a forced draught 
acts on the fuel in a furnace. The thyroid glands, indeed, may be aptly 
compared with the draught-regulating mechanism of a furnace. Excess of 
thyroid leads, as Cramer and Krause (3) showed, to a rapid disappearance 
of glycogen from the liver, and of fats from the body generally. At the same 
time many of the organs are increased in weight and show signs of increased 
activity. The loss of fat may be, and sometimes is, more than balanced by 
the increased weight of some of the viscera. Hoskins explains the hyper- 
trophy of organs resulting from thyroid-feeding as simply due to increased 
metabolism. He points out the similarity which exists between his figures 
and those obtained by Hatai (7) in white rats subjected to a large amount 
of exercise. Hatai kept rats during the daytime in revolving cages which 
necessitated their continual activity. After periods of three to six months 


246 Herring 


the animals were killed, and their organs compared with those of rats which 
had led a more quiescent existence. The long-continued exercise produced 
hypertrophy of the heart, kidneys, and liver to about 20 per cent. above the 
normal. The testes were increased in weight by 12 per cent., the ovaries 
by 84 per cent. The spleen was decreased in weight. Of the ductless 
glands the thyroid showed a diminution in size, the suprarenals were 
unaltered in the male, but hypertrophied by nearly 48 per cent. in the 
female. Changes in the thymus were uncertain. The pituitary body 
was increased in the male by 10 per cent., and decreased in the female 
by 22 per cent. 

My results are similar in most respects to those of Hoskins, but differ 
from his, and from Hatai’s, in the much greater degree of hypertrophy 
reached by certain organs, e.g., the suprarenals, heart, and kidneys, to 
which must be added the pancreas, an organ not specially investigated by 
either Hoskins or Hatai. In respect to the other organs—the liver, 
spleen, testes, and ovaries—our results agree, and it is probable that the 
comparatively small degree of hypertrophy observed in these organs is 
due, as Hoskins surmises, to the generally increased rate of metabolism of 
the body induced by thyroid. It does not seem to me likely that the 
excessive hypertrophy of the heart and kidneys, which in extreme cases 
are more than trebled in weight, can be solely ascribed to a general action 
of thyroid upon metabolism. Such a result may be possible, but as far as 
Iam aware it did not occur in any of Hatai’s rats which were subjected 
to exercise for a considerable portion of their normal life’s span. The 
excess of thyroid in the body of the thyroid-fed animal must upset the 
normal equilibrium which in all probability exists between the internal 
secretions of the ductless glands. Hyperthyroidism stimulates the pro- 
duction of adrenalin. It also produces a striking hypertrophy of the 
pancreas, but whether this is associated with an increase of the islets of 
Langerhans is not decided. The growth of the pituitary body is checked 
in females at any rate, and possibly in males, though the evidence for this 
is doubtful. This disturbance of the relations between the ductless glands 
cannot be ignored, though its significance is not yet capable of being 
thoroughly explained. 

It is probable that in hyperthyroidism many factors are brought into 
play. In this connection the work of Eppinger, Falta, and Rudinger (5) | 
must be cited. These observers claim to have established certain relation- 
ships between the ductless glands. Removal of one gland produces direct 
results on the body from the loss of its secretion, and indirect results from 
the disturbances set up among the others. Between thyroid and pancreas, 
and between pancreas and chromaflin tissue there is a mutual restraining 
influence ; between thyroid and chromaffin tissue there exists a mutually 
stimulating influence. While not committing myself to an agreement with 
the details enunciated by Eppinger, Falta, and Rudinger, I believe there 
is evidence to support some of their views. 


The Action of Thyroid upon the Organs of the White Rat 247 


The enormous heart produced in rats by prolonged thyroid-feeding is 
especially interesting. It is alleged to occur in “ Kropfherz,” and is some- 
times a feature of Graves’ disease in man. Excess of thyroid stimulates 
an excess of adrenalin, but whether the adrenalin takes any part in the 
production of the cardiac hypertrophy is not proved. ‘The administration 
of adrenalin in Graves’ disease is sometimes of great benefit in temporarily 
allaying the cardiac symptoms, slowing and steadying the heart beat, and 
constricting the peripheral blood-vessels. It would appear from this that 
the production of adrenalin is a compensatory phenomenon, its object 
being to overcome the effects of thyroid upon the circulation. In my series 
of rats the animals which showed the fewest symptoms and maintained 
their health most completely during thyroid administration were those in 
which the suprarenals contained the largest amount of adrenalin. The 
suprarenals of male rats respond in this respect to thyroid more readily 
than those of female rats, and it is interesting to note that the males 
appear to be less susceptible to the influence of thyroid than the females. 
In the female rats which showed the greatest susceptibility to thyroid the 
adrenalin of the suprarenals was lowest; in those females which showed 
no adverse symptoms the adrenalin content was very high. It looks, 
therefore, as though a greater secretion of adrenalin is a necessity for 
the maintenance of health whenever there is an increase of thyroid in the 
body. A combination of these factors may be at work in the production 
of cardiac hypertrophy; increased metabolism must also take some part 
in bringing about the condition. 

The hypertrophy of the kidneys appears to be directly related to the 
hypertrophy of the heart, and a close correlation between the two is 
obvious from my figures. It is probable, therefore, that the kidney hyper- 
trophy is dependent upon the same factors as those which bring about the 
cardiac enlargement. 

The constant and great enlargement of the pancreas is chiefly one of 
the glandular acini, as was pointed out by Kojima. It is not necessarily 
associated with an increase in the intake of food, although this commonly 
occurs in rats fed with small amounts of thyroid. It is not unlikely that 
the internal secretion of the pancreas is increased. I have never met with 
glycosuria in rats fed on small doses of thyroid. The increased production 
of adrenalin and the disappearance of glycogen from the liver seem to 
point to a condition tending to produce glycosuria. Possibly this is pre- 
vented by increased activity of the internal secretion of the pancreas. 

Another difficult problem raised is the check in the growth of the 
pituitary in females shown to occur by Hatai as the result of exercise, 
and by Hoskins and myself in thyroid-feeding. In the male no such 
check but rather an accelerated growth of the pituitary is found to occur 
by Hatai, and by Hoskins. My results show an increase in growth of 
the pituitary in males when reckoned in terms of percentage body weight, 
but a decrease in size when estimated in term of percentage body length. My 


248 Herring 


observations, however, are not sufficiently numerous to warrant any criticism 
of Hoskins’ results with regard to changes in the male pituitary, 

No constant changes are demonstrated in the thymus. Its involution 
appears to have been somewhat accelerated in males, and somewhat delayed 
in females in my series of animals, but the changes are slight and may be 
merely accidental. 

The testes show some increase in weight, as also do the ovaries. The 
hypertrophy of the latter is not nearly so great in my animals as in those 
of Hatai’s as the result of muscular activity. 

Iscovesco obtained great hypertrophy of the uterus of the rabbit by 
injections of an ether-soluble extract of thyroid. In only one rat of my 
series can hypertrophy of the uterus be detected, and this is in the largest 
animal of the series. This rat has a comparatively large pituitary body, 
the heaviest suprarenals, and the largest adrenalin content of any animal 
in my series. In the other animals the growth of the uterus is strikingly 
inhibited by thyroid-feeding, and this lack of growth appears to be 
associated with a corresponding lack of growth of the pituitary body. It 
is suggestive of a relation between the two, and the diminished growth of 
the pituitary body may in young animals be the cause of the diminished 
growth of the uterus. 


V. SUMMARY OF CONCLUSIONS. 


Small doses, 0:1 to 0°2 grm. of fresh ox thyroid given daily in food to 
growing white rats, have little effect upon the growth as estimated by 
body length, but tend on the whole to diminish the growth as estimated by 
body weight. In this respect my results confirm those obtained by 
EK. R. Hoskins. 

female rats are more susceptible to the action of thyroid than 
WES. rats. 

Thyroid-feeding causes profound changes in many of the organs of the 
body, and eventually, in the doses mentioned, leads to fatal results. 

Thyroid-feeding stimulates metabolism, leads to the rapid disappear- 
ance of glycogen from the liver as stated by Cramer and Krause, and to 
a general loss of fat from the body as noted by numerous observers. The 
diminution in body weight caused by the loss of fat may for a time be 
counterbalanced or even more than counterbalanced by the increased weight 
of many of the viscera. 

My results agree with those of Hoskins in that thyroid-feeding is 
found to produce enlargement of the suprarenals, heart, liver, spleen, testes 
and ovaries; no constant changes in the thymus, and a decrease in size of 
the pituitary body in female rats. They also confirm Kojima’s statement 
that thyroid-feeding produces hypertrophy of the pancreas. 

The thyroid glands of thyroid-fed animals are decreased in weight, and 
show signs of relative inactivity. 

My results differ from those of Hoskins mainly in the greater extent 


ss a ae 
—_ 


The Action of Thyroid upon the Organs of the White Rat 249 


of hypertrophy shown in my series of rats, more especially of the supra- 
renals, heart, kidneys, and pancreas. The heart and kidneys may be more 
than trebled, and the suprarenals and pancreas more than doubled in 
weight by thyroid-feeding. 

The enlargement of the liver, spleen, testes, and ovaries may be 
ascribable, as Hoskins suggests, to the general increase of metabolism in 
the body. But the greater enlargement of the suprarenals, heart, kidneys, 
and pancreas points to something further than this. There is an increased 
production of adrenalin by the suprarenals of thyroid-fed animals, and the 
maintenance of health appears to be bound up with the attainment of a high 
adrenalin content in such animals. It is suggested that the adrenalin output 
is compensatory to the thyroid, more especially in its action upon the circula- 
tion. The combination of these factors along with the increased metabolism 
may be the cause of the great hypertrophy of the heart and kidneys. 

The hypertrophy of the pancreas may be concerned in regulating 
carbohydrate metabolism, which would otherwise be upset by the increased 
production of adrenalin. 

The uterus of the young rat appears to be checked in its growth by 
thyroid-feeding. The small uterus is associated with a similar check in 
the growth of the pituitary body, and the idea suggests itself that there 


_ is a close correlation between these two organs. 


I have to thank Mrs Donald Mills for her help in looking after the 
animals during the last part of the work. To Mr Niven of Strathkinness 
I am indebted for a plentiful supply of fresh ox thyroid. 

The expenses of the research have been met by a grant from the 
research fund of the Carnegie Trust for the Universities of Scotland. 


VI. LITERATURE CITED. 


(1) Brept, Innere Sekretion, Wien, 1913. 
(2) Bircuer, Med. Klinik, 1910, 391. 
(3) Cramer and Krauss, Proc. R. Soc., B, 1913, Ixxxvi. 550. 
(4) Donatpson, The rat, Wistar Institute Memoirs, 1915. 
(5) Eppincsr, Fara, and Ruprneer, Zeitschr. f. klin. Med., 1908, Ixvi. 1. 
(6) GupernatscH, Amer. Journ. Anat., 1913-14, xv. 431. 
(7) Harat, Anat. Rec., 1915, ix. 647. 
(8) Hara, Amer. Journ. -Anat., 1913, xv. 87. 
(9) Herrine, Quart. Journ. Exper. Physiol., 1916, ix. 391. 
(10) Herrine, Quart. Journ. Exper. Physiol., 1917, xi. 47. 
(11) Hosxrys, E. R., Journ. Exper. Zool., 1916, xxi. 295. 
(12) Hoskins, R. G., Journ. Amer. Med. Assoc., 1910, lv. 1724. 
(13) Iscovesco, Compt. rend. d. 1. soc. d. Biol., 1913, Ixxv. 361. 
(14) Jackson, Amer. Journ. Anat., 1913, xv. 1. 
(15) Kine, Anat. Rec., 1915, ix. 751. 
(16). Kosmma, Proc. R. Soc. Edin., 1916, xxxvi. 240. 


250 


(17) Livineston, Proc. Soc. Exper. Biol. and Med., 1914, xi. 
Hoskins (11). 


Herring 


(18) Moussu, Compt. rend. d. 1. soc. d. Biol., 1899, li. 
(19) ScHArER, Quart. Journ. Exper. Physiol., 1912, v. 203. 
(20) Swate Vincent, Internal secretion and the ductless glands, London, 1912. 
(21) Urrerstrom, Arch. de méd. expér., 1910, xxi, p. 550. 


VAT. 


TABLES. 


Quoted from 


TABLE I.—AVERAGE WEIGHTS AND PERCENTAGE CHANGES IN ORGANS PER 100 GRM. 
Bopy WEIGHT FROM Rats oF TasuEs III., [V., VII., anp VIII. 


Organ. 


Suprarenals . 


Hearty. 
Kidneys 
Pancreas 
Liver 
Spleen . 
Testes . 
Thymus 
Ovaries 
Uterus. 
Thyroids 
Pituitary 


Male. 
Control. ay ay Percentage | Control 
ed. change. 
0-016 0-025 + 56 0-029 
0°412 0°922 +123 0523 
0-747 1545 +106 0°831 
0°361 0-763 sale 0°467 
3°534 5139 + 45 4391 
0:469 0°654 + 39 0°579 
1157 1253 ae 8) 
0240 
0-041 
0168 
(0:0145) | (0:0103) | (—29) 0:0137 
(0:0038) | (0:0045) (+18) 00052 


Female. 


Thyroid- 
fed. 


0-041 
0°834 
1361 
0°842 
5°762 
0-712 


0°259 
0-046 
0°143 
0:0104 
070046 


Percentage 
change. 


+41 
+59 
+63 
+80 
+31 
+ 22 


am Ul 
+12 
—14 
—24 
—ll 


The figures in parentheses are taken from another series of male rats. 


TaBLE IJ].—AVERAGE WEIGHTS AND PERCENTAGE CHANGES IN ORGANS PER 100 MM. 
Bopy LenetH From Rats oF TaBuEs III., IV., VII., anp VIII. 


Organ. 


Suprarenals . 


Heart . 
Kidneys 
Pancreas 
Liver 
Spleen . 
Testes . 
Thymus 
Ovaries 
Uterus. 
Thyroids 
Pituitary 


Male. 
Control. oe 
fed. 
0015 0:023 
0397 0°860 
0°720 1°442 
07348 0°712 
3°404 4°796 
0°452 0-611 
1114 1170 
(0:0124) | (0:0064) 
(0:0033) | (0-0027) 


Percentage 


change. 


2a GS) 
+116 
+100 
+104 
+ 40 
+ 35 
ans 


(— 48) 
(— 18) 


Female. 
‘ Thyroid- 
Control. ead 

0:020 0:029 
0°355 0°588 
0°564 0-960 
0°317 0:594 
2°983 4-063 
0°393 0°502 
07163 0182 
0-028 0:032 
0-114 07101 
0:0093 0:0073 
0:0035 0:0032 


Percentage 
change. 


+45 
+65 
+70 
+87 
+36 
SE 7/ 


+11 
+14 
—ll 
—21 
— 8s 


The figures in parentheses are taken from another series of male rats. 


The Action of Thyroid upon the Organs of the White Rat 


TABLE IIIJ.—ContTrRot, MALE RATS FED ON BREAD AND MILK. 


No. 1 


In ADDITION 0°2 GrM. FRESH OX FLESH DAILY. 


251 


RECEIVED 


Bs) Se.) Aso eye 8. y Se eo a 
eke fee (ea mens |B) dis |3 |3 
ofits 2 |8|s |] se | & B= wh | & “ Z 

@ 1 se2\qh| = &|.2 | Ss 05 So | ‘ot | wy Sm | Sh] 2 & 

SiSS5\|Seiaeciecl & 29 | 24 2 ¢ oo | Sa |s8elaad 

Z\selar|s"|5) | Gee | me | es | es | PR | SA | eer 
sem |e es C5 ® ‘oo — ‘SD = > 2 — 
AgZ|c we TS = ie S = 

1 42 | 88 154 66. 184 | 0-028 0-780  1:224 | 0°566 | 6317 | 0°744| 1:828 

2 | 165 192 | 0°025 | 0°736 | 1°397 | 0°643 | 7:078 | 0:903| 1°77 

3 170 184 | 0-026 | 0-702 | 1:266 0-637 5:415  0°568 2-085 

4 | | 173 184 | 0:030 | 0°704 | 1:986 | 0°690 | 6:295 | 1°033)| 2°306 | 

5 | 180 185 | 0:097 | 0-784 | 1°126 | 0585 | 5:295 | 1:013)| 2°311 

6 | 200 193 | 0°027 | 0°754 | 1°388 | 0°576 | 5°803 0°809 | 2°406 

7 | | 240 212 | 0°038 | 0°825 | 2:091 | 0941 | 9-078 0°948 | 2°123 

Se Sy = é _ 

Averaye gross \ | 195 | 190 | 0029 | 0°755 1:368 | 0°662 | 6-468 0859) 2-118 
weights. | 

rm. fin: ; ; Lacan aa 

Average per 100grm. final body || 9.916 | g-412 | 0-747 | 0°361 | 3-534 | 0°469| 1-157 

weight E é eee ss res ia Pere: 

Average per 100 mm. finallength| 0015  0°397  0°720 | 0°348 | 3-404 07452) 1°114 
TasLe [V.—Ma.e Rats FED ON BREAD AND MILK WITH THE ADDITION OF 

0-2 GrM. FRESH Ox THYROID EACH DAILY. 

9) eRe a ee a a 8, oy [i0; /\. tt. | 12+) a3; 

' a — e a > eB = =| mn 
pa a | io +. || wos | a =| fal = ia | aa ee RS a >) cea pa 4 Oo . 
eet pe eee, Si) eal gel alae: epee ee) ace lise Ae ae | 
a |SelqgalFa\ S| 5 | S2/Sh|\s& Sa) fm) th/ Se 
zm |s2\ma|25| 2) © | Gs | SS | Sa ee | me | Se | ee 

Bee |k TH 2 | ‘oo 2 T = © | 'o “OD 

a ae rf Fas cas = iy s| = Wage 
1 37 | 147 | 216 | 69 | 208 | 0:046| 1°578| 2827) 1°742.|11°277 | 1:632| 2-477 
2 38 | 119 | 197 | 78 | 202 | 0:037] 1°340| 2°162 > 1°600| 7-400 | 1:161| 2°335 
3 39 937) 163) Go Lola OOsoim Alot ola.) W207 | 6601) | 1083.1, 2-72 
4 40 106 | 168 62 | 190 | 0:049| 1:348| 2:160' 1:238| 7-902 | 0:991)| 2-232 
5 44 | 109 | 177 | 68 | 192 | 0:046/| 1°884| 3:064) 1°349| 8:922-| 1:245)| 9:244 
6 45 | 117 | 173 | 56 | 197 | 0°053) 1°730| 2°547 | 1:370)} 8:008 | 0°821)] 2°380 
yA bl 114 | 182 | 68 | 189 | 0:058 2°455 4°758 1°212|15°302 | 1313 2°135 

Average 42 | 115 | 182 67 | 195 | 0°046| 1678) 2°813 1:°389) 9°354 1192 | 2-282 

Average per 100 grm. finalbody weight) 0°025) 0°922 1°545 | 0°763) 5139 0°654 = 1253 

Average per 100 mm. final length 0:023| 0860 1:442 0-712 | 4°796 | 0611! 1-170 

| | | 


252 


Herring 


TapLE V.—Mate Rats oF SAME AGE FROM TWO CLOSELY-RELATED LITTERS. 
a, Controls fed on bread and milk with 0-2 grm. fresh ox flesh each daily. 


Average per 100 grm. final body weight 


Average per 100 mm. final length 


1 2. 3. 4. dD. 6. 
ees Original) Final Gissy. eset 
7: An 3 : g 
Number. periment weight | weight in grm. | in mm. 
: “| in grm. | in grm. 
in days. 
1 24 80 145 65 184 
2 26 75 144 69 184 
3 26 74 148 74 187 
Average . 25 76 146 69 185 


hs 
Weight 
of supra- 
renals 
in grm. 
0024 
0:025 
0-025 


0°025 
0-017 


0-013 


8. 


Weight 
of heart 
in grm. 


0°729 
0°675 
0-706 


0°703 


0-481 


0°380 


9. 


Weight 
of spleen 
in grm. 


0°899 
0°699 
1044 


b. Fed on bread and milk with 0-2 grm. fresh ox thyroid each daily. 


4 24 

} 5 26 
6 26 
Average . 25 


86 


Average per 100 grm. final body weight 


Average per 100 mm. final length 


Number. 


il 


2 
3 
+ 


Average . 


0-060 
0-046 
0:032 


0-046 


0:031 


0-024 


TaBLeE VI.—Mate Rats FROM SAME LITTER. 
a. Control fed on bread and milk with 0-2 grm. fresh ox flesh daily. 


2 


= 


Duration 
of ex- 
periment 
in days. 


8 


3. 
Original 


weight 
in grm. 


76 


4. 
Final 


weight 
in grin. 


114 


Average per 100 mm. final length 


Average per 100 grm. final body weight 


5. 


Gain 
in grm. 


38 


6. 


Length 
in mm. 


164 


de 


Weight 
of supra- 
renals 


in grm. 


0-019 


0-015 


0-011 


1-704 
1:259 
1298 


1°420 


8. 


Weight 
of heart 


in grm. | 


b. Fed on bread and milk with 0°2 grm. fresh ox thyroid each daily. 


8 
8 
8 


8 


70 
73 
62 


68 


98 
98 
89 


95 


Average per 100 grm. final body weight 


Average per 100 mm. final body length 


145 
158 
148 


150 


0-027 
0°026 
0:023 


0-025 


0:026 


0-016 


0°893 
07845 
0°848 


0-862 


0-907 


0-574 


9. 
Weight 


of spleen 
in grm. 


1:056 


0-926 


0°643 


1538 
Ics iz 
1:296 


1-402 


1475 


0°934 


bo 
t 


The Action of Thyroid upon the Organs of the White Rat 53 


Taste VIJ.—Conrrot FemMate Rats FROM THE SAME LITTERS AS THE Rats IN 
TaBLE VIII, rep oN BREAD AND MILK. 


1 2/3) 4/6) 6 | 7 8 Oe VG eT bs els. ad] Teel) Py. | 

, wie ug fe re > a 5 4 |8 |o |g | 
I Pale sierd| eto eee a leo | Oa baa | Be Alo ls ol la 
meer SiP ars 8) Ml ae | moi a | 4S | ea les Ej col oo) eb) ol 
- = Be Bi> el gt El ag ok aH ra Cn |) Se me a8 ge giXragiga 
= Cea eT we | SO] aw WD] SO BO} 008 | 4 bn | on SRO Ro Ba. 8 /o 8 
= Secisaias|d |Bleagisa#ae lsaqal Fo Sa lba le Sle sieqgisatsn 
| |S ee isis | 2 | og | em | tom ES | BA | tom Pele lea [oo 
SES & oO] sil sei | Gis |S fs [so to |P [os 
* | ot) a ae | a. iva oer coal 5) 
1 21 | 70 | 93) 23 | 143) 0-028) 0°636 0°848 0-436) 4°986| 0:°838/163/ 49 | 131] 19-3 4°] 
2 28 | 72 |118| 46 162) 0:035| 0°575 0-980! 0-408 4828 | 0°64y/ 332| 49 |214| 17°5| 6-5 
3 29 | 56 106 | 50 165 | 0°031 | 0°574 |) 0°937 0563 5°018)| 0°402| 285) 51 | 202! 15-1)! 6-8 
4 30 | 73 | 81) 8 | 141} 0-025) 0-430) 0°727 | 0-486 3°913| 0°551/136| 25 | 64| 13:3 5:0 
5 34 | 83 /111| 28 | 161} 0:029 0-474) 0°925| 0-490 4:036| 0°509| 420| 31 | 95] 19-8 52 
6 36 | 83 |} 125} 42 158 | 0°037 | 0°648 | 0°914) 0°677  5°215| 0°825/ 255 | 59 |170| 13-1! 6-4 
7 46 70 | 110 40 160 | 0:031 0549 0°837 | 0°415 4°587) 0528/194! 44 | 372| 18-0! 5-4 
Average| 32 | 72 |106| 34 156] 0:031| 0°555 0881 0-496 4655 | 0614/2955, 44 |178| 14-6) 5-6 
Average per 100 grm. a et i } ; e Peale roe a 
body weight ; } ne Re 029] 0°523) 0°831 | 0°467) 4:391 | 0°579| 240) 41 | 168} 13°7| 5:2 
Average per 100 mm. final) | ,... Pee OPS 21 9. bers re Pe ae 
body length ( 0020} 0°355 | 0°564) 0°317| 2:983 | 0°393 | 163 28 |114| 9-3) 3-5) 


SN  ——————————————————————————— 


TaBLe VIII.—Femate Rats FROM THE SAME Lirrers as THE ContRoL Rats IN 
TaBLe VII., rep oN BREAD AND MILK WITH THE ADDITION OF 0'1 GRM. OF 
FresH Ox THYROID DAILY. 


1 2 +3 4 5 6 if 8 9. 10 11 12 1S.) 045) 5. |) 16) 17 

sy 8 7S) Vets tal Wateteret || SIAR Boe ee ees 
Peele | Fle) ea Sle. (se @|5 16. b le 18 fe ee 
g SSlFaS Ee] la | 2a lek | Se] 2S las | 2 Bw ob F eb) Seb | OiA 
= Sela le & S/ =| SR | Sh | 8 & "oo2 | oh | Sa (So S/S 8/6 Bin a 10.8 
Peeege dad 2) | Sa \Se (Sel 22 lea lea Ssleasigqieqiss 
A Bele |e “|-a | 2 | eos | 27 | or | 3 4 | "orn feo lon \eo | "ep (a0.8 
aote i 16 |8|38 3 2 is |e |e ‘3 {2 is 

Ag\6 | Hilfe |e |e See Se. i eee 

1 21 | 72 | 73; 1 |120} 0-040] 0°832| 1°320| 0°454| 5657] 0°517| ... | 25 55] 7-6 | 3-7] 
2 28 | 76 | 99} 23 | 155} 0:040} 0°835| 1:257| 0-846 | 5-760] 0:358| 402! 46 631/13°3 | 5-0! 
3 29 | 71 |106/ 35 | 164/ 0°035/ 0°801/ 1:290) 1171 5°318! 0:495| 498] 39 95/12-0 | 4:0 

4 30 | 73 | 88] 15 | 140} 0:041| 0°854} 1:333] 0-808) 4:943 0-910! 91] 28 71 Ol | 4:2 
5 34+ | 95 | 127} 32 | 165] 0:044| 0-808] 1-587 0°945 | 6469 0°724/324| 47 89/108 | 4:4 
6 36 | 86 |140| 54 |174| 0:074| 1111} 2:036| 1-263) 8-450 1:172/171|104 496/14-0 | 8-6! 

7 46 | 72 | 138] 66 |174| 0:°046, 1:187| 1-665 1006, i 779 | 1°312|227| 71 287 |13°7 | 61 
enn ee ee ns — W—_eoo__ TT OO EE CO ees eS 
Average| 32 | 78 |110| 32 |156 0-046 0°918/ 1:498| 0:927 6339 0°784/285| 51 158/11°5 51 


Average per 100 grm. final ; ' - 
body weight : , 0-041. 0°834| 1°361 O82 5°762 | 0°712| 259) 46 143)|10°4 | 4°6 


Average per 100 mm. aa ; : : : i 
body length. ; | 0:029 | 0°588 | 0-960 | 0°594 | 4:063  0°502 ‘182 32/101) 7-3 | 3:2 
0 ee ca ee ee A Ne Rd |e 


—__, 


STUDIES ON THE ENDOCRINE GLANDS:—PAPER L: 


THE 


RELATIONS BETWEEN THE PANCREAS AND THYROID 
AND PARATHYROID GLANDS. By Masanaru’ Koma, 
Fleet Surgeon, Imperial Japanese Navy.! (From the Physiology 
Department of Edinburgh University.) (With twenty-nine figures 


in the text and two coloured plates.) 


(Received for publication 1st December 1916.) 


CONTENTS. 


MeErHops OF INVESTIGATION 
Preparation of extracts 
Diet and treatment 
Operative procedures 
Microscopic methods 
EFFECTS UPON THE RaT OF RemovaL OF ‘THYROID AND PARATHYROD, AND OF 
ADMINISTRATION OF THYROID AND PARATHYROID 
Microscopic appearances of pancreas of rat— 
(a) in the normal animal 
(b) after parathyroidectomy 
(c) after thyroidectomy 
Effects of thyroid feeding— 
(a) in the unoperated animal 
(b) after parathyroidectomy 
(c) after thyroidectomy 
Effects of parathyroid feeding— 
(a) in the unoperated animal 
(b) after parathyroidectomy 
(c) after thyroidectomy 
Thyroid feeding in different sexes 
Thyroid feeding during varying periods 
Thyroid feeding with intermission, and also with i increasing doses 
Effects of thyroid feeding on weight of pancreas 
Effects on the amount and aca of the urine . 
Small doses of thyroid a 
Prolonged feeding with thyroid . 
Feeding with water-extract of.thyroid and with its residue 
Feeding with alcohol- and ether-extracts of (aan and with the residue after 
such extraction 
Subcutaneous administration of thyroid extract: 


259 
264 
264 


265 
268 
269: 


273 
273 
274 
274 
277 
279 
280 
281 
284 
285 
287 


289 
290 


‘ The author desires to record his acknowledgments to Professor Schafer for assistance 
and advice in carrying out these investigations, The expenses have been defrayed by 
grants from the Earl of Moray Fund for the Endowment of Research in the Univer-ity of 
Edinburgh, and from the Carnegie Trust for the Universities of Scotland. A preliminary 
account of some of the results was communicated to the Royal Society of Edinburgh on 


July 3, 1916, and is published in the Proceedings (1). 


VOL. XI., NOS. 3 AND 4.—1917. 17 


256 Kojima 


PAGE 
THYROID FEEDING OF CASTRATED RATs : : 2 : 290 
Does castration alone affect the structure of ihe! pancreas ? : 291 
Does previous castration affect the results of thyroid feeding on the p: ancreas ? 291 

DoEs THE ADMINISTRATION OF COMBINATIONS OF IODINE WITH PROTEIN AFFECT 

THE PANCREAS IN THE SAME MANNER AS ADMINISTRATION OF THYROID 
SUBSTANCE ? 293 

WHatT IS THE Berner UPON THE Pinel OF ADMINISTERING loprore AND 
OTHER SALTS ? ; . ; : : : : ; 294 
Sodium iodide. : : 3 : : : ; ; 294 
Potassium iodide . : 4 f 5 298 
Sodium bromide, sodium chloride, and sodium tluoride : ; 300 
Sodium carbonate, sodium phosphate, and sodium sulphate. ; t 300 
EFFECTS OF ADMINISTERING CERTAIN HORMONIC SUBSTANCES : ; ‘ 302 
Pilocarpine : : : : : : ; ; 302 
Adrenalin 3 . fk : ) : 302 
Pituitary body . E ; : é : 303 
(a) Anterior lobe. ; ; . 303 
(b) Posterior lobe. : : 303 
EXPERIMENTS ON THYROID FEEDING OF Arum. OTHER THAN are : : 306 
Mice ; : ; : : ; : : ; ; 306 
Cats and dogs : : : ; : 4 : : 307 
Rabbits and aes pigs . : , ; : 310 
Birds : ‘ : : : 4 ; ; 5 311 
GENERAL SUMMARY . s : 3 3 3 : : ? 314 


METHODS OF INVESTIGATION. 


THESE investigations had for their original object the determination of 
the effects of extracts of certain of the endocrine glands upon other organs, 
but in this paper the observations deal for the most part with the relations 
between the thyroid and parathyroids and the pancreas, the experiments 
having been chiefly designed to observe the effects of thyroidectomy, of 
parathyroidectomy, and of feeding with thyroid and parathyroid upon 
the pancreas. 


Preparation of Extracts. 


The glands and gland-extracts which have been employed for feeding 
are the thyroid of the ox and sheep, and the parathyroid of the ox. In 
some of the thyroid-feeding experiments fresh glands were employed ; in 
others dried material was used. This was prepared in the following way :— 
Fresh glands from the ox, obtained direct from the slaughter-house, were 
freed from the surrounding tissues. They were then minced, and the 
material placed in a thin layer upon a glass plate, on which it was dried in 
an incubator at 37° C. The dried material was ground up with a mill, and 
was then kept in an exsiccator. Extracts of this or portions of the gland 
substance itself were added in a small but definite amount to the food of 
the animal: this food was of fixed quality throughout. For parathyroid 
feeding, glands of the ox which had been collected and preserved for a 
time in chloroform were employed, the glands being minced and dried in 
the manner above described. The animals used for feeding experiments 
were for the most part white rats, generally adult, i.e. sexually mature, 


Studies on the Endocrine Glands 257 


although varying somewhat in size. For certain experiments other animals 
(dogs, cats, rabbits, guinea-pigs, fowls) were employed. 


Diet and Treatment. 

Animals of about the same size, and generally of the same sex, were 
chosen and fed ad libitum with a diet of fixed and known composition 
for a week or more. They were weighed at frequent intervals. Only 
those that showed during this preliminary period no marked difference in 
weight were employed for the actual experiment. The food used for dogs 
and cats was meat, oatmeal porridge and milk: for guinea-pigs, rabbits, and 
hens—oats, cabbages, and Indian corn. As the standard food of the rats 
rusks! were eventually chosen. They were ground in a small mill, and 
about 40 per cent. water was mixed with the powder so as to make it into 
a soft paste. The composition of the rusks was as follows :— 


Protein ; , ae 2 
Glucose ; : . 2°98 
Fat ’ : . : : ; 6°63 
Cane sugar. ’ . 2452 
Calcium ae i005 
Starch . : : . 48°92 
Water . : : ; . 1635 
Ash. . | 86 


Dried residue, obtained after filtering off 
the glucose which had been obtained in 
carrying out the estimation of starch . 8°58 


100-00 


It will be observed that this furnishes a diet with the food-stuffs in 
suitable proportions. Other foods which were at first tried were found to 
be too variable in composition or otherwise unsuitable. Additional water 
was offered to all the animals once or twice every day, but as a general 
rule it was found that the rats had a sufficient amount in the paste 
supplied to them. 


For investigating the effects of operation or diet upon their metabolism 
the rats were kept—in groups—in the small wire metabolism cages de- 
scribed by Professor Schafer (Quart. Journ. Exper. Physiol. 1912, 
vol. v. p. 204). Occasional. CO, estimations were made in certain of the 
experiments. The effects upon metabolism were not investigated in the 
other animals employed. 


Operative Procedures. 


Upon some of the animals certain operations were performed. 
(a) Thyroidectomy.—After being anzsthetised with ether, sometimes 
combined with chloroform, a median incision was made over the upper part 


1 Those made by Habbard of Glasgow. 


258 Kojima 


of the trachea and the lower part of the larynx, and the thyroid gland 
having been exposed, a double ligature was laid between the lower end of 
the gland and the adjacent tissue on each side. A cut having been made 
between the ligatures, the whole gland, including the isthmus, was quickly 
removed with the aid of fine scissors, great care being taken not to injure 
the neighbouring nerves. After removal of the thyroid, the attached para- 
thyroids—which in the rat are single on each side and usually external to 
the thyroid and attached to its border—were sought for with the aid of a 
lens and, when found, removed and replaced in the depths of the wound, 
which was then sewn up. The operation was performed aseptically ; any 
bleeding which occurred was controlled by pressure with sterile cotton- 
wool soaked with dilute adrenalin. The exterior of the wound was painted 
with tincture of iodine and completely covered with collodion. 

In all, fourteen adult male rats were subjected to thyroidectomy ; but of 
these some died shortly after, and apparently as a result of, the operation. 

(b) Parathyroidectomy.—The thyroids having been exposed and 
the parathyroids found, the latter were snipped away with small curved 
scissors along with a minute portion of thyroid substance. The fact of 
their removal was determined by examination with a lens, but to ensure 
that the removal was complete the thyroids, after the animal was killed 
at the end of the period of observation, were cut completely in series; in 
no case was any trace of parathyroid found. 

Six rats were subjected to parathyroidectomy. One of these died five 
days after the operation; the others were used for experiment and 
observation. 

(c) Castration—This operation was performed in the usual way by 
an incision through the scrotum and ligature of each cord, the testicles 
then being removed. 

Hight adult ‘male rats were castrated. All rapidly recovered from 
the operation. 


Microscopic Methods. 


At the end of the period of experiment, the animal—or group of 
animals—having been killed with chloroform, the organs to be investigated 
were placed at once in 10 per cent. formol. The pancreas was always 
freed from mesentery and from the spleen, and if small, as in the case of 
the rat, was placed as a whole in the formalin solution; if larger, as in 
the dog and cat, it was first cut into a number of thin pieces. After the 
fixation had been completed by formol the portions of organs to be 
investigated were usually embedded in paraffin and thin sections (about 
lu) were cut from them. ‘The staining methods employed were (1) 
hematoxylin-eosin, (2) Mallory’s stain (acid fuchsin, aniline blue and 
orange G), and (8) Muir's stain (alcoholic eosin and methylene blue). 


Studies on the Endocrine Glands 259 


EFFECTS UPON THE RAT OF REMOVAL OF THYROID AND PARATHYROIDS; 
AND OF THYROID AND PARATHYROID ADMINISTRATION. 


During a preliminary period a diet consisting of a mixture of minced 
lean meat and ground rusks in the proportion of 60 per cent. and 
40 per cent. respectively had been employed, a sufficient amount of water 
being added to make the mixture into a soft paste. But subsequently 
{at the commencement of thyroid and parathyroid feeding) the meat 
was omitted, rusks and water having by then been definitely adopted 
as the standard diet. 

The rats were divided into three groups, A, B, and C. Group A con- 
sisted of five normal male adult animals and was used as a control: group 
B of five parathyroidectomised males (operated upon February 8, 1916); 
and group C of three thyroidectomised males (operated on February 9, 
1916). The animals belonging to each group were kept in the same cage, 
and all three cages were close together in the same room and under the 
same conditions of temperature. No special symptoms appeared to be 
produced either by the parathyroidectomy or the thyroidectomy during 
the period of observation.! On March 24 one rat in each group was 
killed and examined. This animal will be called No. 1. 


Microscopic Appearances of the Normai Rat’s Pancreas 
and the Changes produced by Parathyroidectomy and 
by Thyroidectomy. 


No. 1 rat of A group (unoperated). Weight of animal 255 grm. All 
the organs are normal. 


(a) The Normal Pancreas. . 


The pancreas has been examined not only in rat No. 1 of A group, but 
in a large number of other normal rats, both male and female (pregnant 
and non-pregnant), young and full-grown, so that the following description 
of the normal pancreas is a general one. 


Under the microscope sections of the normal rat pancreas 
(figs. 1-5 and Plate I., A) show alveoli which are approximately equal 
in size. Most of the cells are rounded-angular and are also of fairly 
equal size, although there are a certain number of small cells inter- 
mingled with the others. The outer zone of the cells is usually 
narrower than the inner: it exhibits striations when carefully 
examined under a high power. It is stained by hematoxylin more 
deeply than the inner zone, and it is also stained of a faint blue 
colour by Mallory’s and Muir’s stains. The inner zone is relatively 
wide, occupying the greater part of the cell. It is only very faintly 
stained by hematoxylin alone; its granules are coloured by eosin. 
The cell-nuclei do not generally vary greatly in size (5u to 6°2u), 


1 It is well known that rats resist the effects of total thyroparathyroidectomy for a 
prolonged period. 


260 Kojima 


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Fic. 1.—Section of normal pancreas of rat (male), rusk-fed. Microphotograph ; magnified 
90 diameters. Hematoxylin preparation. 
The alveoli show a thick inner or zymogen zone, which appears light in the 
preparation, and a very thin outer zone, with nuclei lying between them. A section 
of an islet is seen in the middle of the photograph. 


Fic. 2.—Part of a similar section ; magnified 400 diameters. Hematoxylin preparation, 
This exhibits the alveolar cells and their nuclei better than fig. 1; it shows the 
thinness of the outer zone and the large amount of zymogen in the inner zone of 


the alveolar cells. 


Studies on the Endocrine Glands 261 


although in young rats of both sexes (fig. 3) there is a more 
considerable variation (3'7u to 8u); even in these there are few 
smaller than 5u, although occasionally a nucleus is seen as large as 
10u. The nuclei are stained blue by Mallory and Muir, and very 
distinctly by hematoxylin. The nucleus usually has a clear appear- 
ance owing to the comparatively few and fine chromatin granules it 
contains. The nucleoli are stained much in the same way as the 
chromatin of the nucleus, but with Mallory and Muir they sometimes 
appear red (Plate L, A). In some cells the nucleoli are large, and occupy 
the middle of the nucleus: there is then a tendency of the chromatin 
granules of the nucleus to be disposed near its periphery. The nuclei 


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Fic. 3.—Section of pancreas of young rat (male). Microphotograph ; magnified 
400 diameters. Hematoxylin preparation. 

If this figure is compared with fig. 2, which is from the gland of an 
adult animal, it will be seen that the chief difference lies in the fact that 
the nuclei are much more variable in size; it is especially obvious that 
there are some of very large size. 


are situated between the outer and inner zones of the cytoplasm. 
Zymogen granules are abundant in all alveoli, filling almost the 
whole of the inner zone (figs. 4 and 5). They are coloured purple-red 
or rose-red by Mallory and Muir (Plate I., A); in the hematoxylin- 
eosin preparations they are stained by eosin. In photographs they 
come out black. In well-developed cells they form a large mass 
within the cytoplasm. In small cells and in the cells of the alveoli 
which are near the surface of the organ there is a tendency for the 
granules to be more scattered in the cytoplasm and less accumu- 
lated. Zymogen granules are always particularly plentiful in the 
cells of those alveoli which are immediately adjacent to the islets 
of Langerhans. 

In pregnant animals there are relatively fewer zymogen granules 
(fig. 6); this is most marked in the earlier stages of pregnancy. 


262 Kojima 


Fic. 4.—Section from a normal rat-pancreas like that shown in fig. 1, but 
stained by Mallory’s method (acid fuchsin, orange G, and aniline blue) 
instead of hematoxylin. Microphotograph ; magnified 100 diameters. 

‘The zymogen masses, which are stained deep red by the acid fuchsin, 
come out black in the photograph. An islet of Langerhans is included 
in the field: zymogen is most abundant in the cireumjacent alveoli. 


Fic. 5.—Section of the same pancreas, also stained by Mallory’s method 
but magnified 500 diameters, 
The zymogen granules come out black in the photograph. Some of 
the nuclei of the cells are faintly shown. 


Studies on the Endocrine Glands 263 


There is no appreciable difference in the quantity of the granules 
as a result of twenty-four hours’ fasting. 

A few small alveolar cells are stained homogeneously yellow by 
the orange G of the Mallory stain ; indeed, the nuclei are coloured blue. 
There is rarely or never evidence of karyokinesis in any cell of the 
normal pancreas. I have never been able to detect a mitosis, even 
in the pancreas of animals which are still growing. Occasionally 
vacuoles occur in some of the cells of the alveoli, but they are never 
so large and conspicuous as the characteristic vacuoles which make 
their appearance as the result of feeding with thyroid (see p. 267). 


Fic. 6.—Section of pancreas of pregnant female rat. Microphotograph ; magnified 
140 diameters. Mallory’s stain. 

It will be noticed that there is a general diminution of zymogen granules 
(compare with fig. 4). An islet is present in the middle of the section; the 
diminution of the zymogen granules extends also to the alveoli round this, which 
under ordinary conditions are very full of zymogen. 


The connective tissue of the gland is coloured an intense blue by 
Mallory. It surrounds the alveoli and lies between the islets and 
the alveoli. 

The centro-acinar cells are spindle-shaped, with small nuclei 
exhibiting a faintly granular chromatin. ‘The size of these nuclei 
is from 24 to 2%5u. In Mallory-stained sections there is sometimes 
seen within the ducts a red material coloured homogeneously—no 
doubt a coagulum from the secretion. 

The islets form a compact mass of cells with irregular sinus-like 
capillaries between them. The cells of the islets are of nearly equal 
size. They contain scattered granules much finer than the zymogen 
granules of the alveolar cells, and staining red by Mallory and Muir. 
The nuclei are stained faintly blue by Mallory. They are fairly 
uniform in size, measuring about 5u. 


264 Kojima 


(b) Effects of Parathyroidectomy on the Rat’s Pancreas. 


No. 1 rat of B group. Parathyroidectomised thirty-five days before 
death. Weight 295 grm. 

There is nothing particular to record post-mortem. The thyroid gland 
is perhaps somewhat paler in colour than normal. The intestines and other 
organs seem to be normal. 


Pancreas.—The nuclei of the alveolar cells measure about 5u; a 
few are a little larger (6:2u). These last contain distinctly outlined 
nucleoli, whilst most of the others have uniformly scattered chromatin 
granules, and the nucleoli are not distinct. When the nucleolus is 
marked there seems to be a clear space around it in the nucleus, 
the particles of chromatin being for the most part arranged near 
the periphery. In the Mallory preparations the nucleoli are fre- 
quently stained red, but sometimes faintly blue: the rest of the 
nucleus takes the blue colour. Speaking generally, the alveolar 
cells are similar to those of the control, but the cytoplasm has a less 
compact appearance and contains fewer zymogen granules, the outer 
zone being a little thicker in proportion to the inner. Some alveoli 
show vacuoles—but these have not the sharp outlines and conspicuous 
appearance of the vacuoles met with in the thyroid-fed animal (p. 267). 

The islets of Langerhans have a somewhat looser texture than 
usual, but there is a certain amount of difference in this respect 
even in normal animals. No difference from the normal can be 
determined in the centro-acinar cells, nor in the connective tissue of 
the gland. 


Two parathyroidectomised rats, which had been fed during the whole 
time with ground rusks and water alone (except for thyroid additions during 
the second week and parathyroid additions during the third week), were 
killed on May 15, ninety-seven days after the operation. These rats are 
respectively Nos. 4 and 5 of group B. The weight of No. 4 was 120 grm.: 
of No. 5, 110 grm. The original weights were 125 grm. and 135 grm. 
respectively. Post-mortem there is nothing noticeable. 


Pancreas—tThe alveoli are moderately large; their cells are for 
the most part large, but there are many smaller cells, the nuclei of 
which are relatively small and situated close to one another, as if the 
result of division. The diameter of the nuclei varies between 5u 
and 6°2u, but some are considerably larger. Nucleoli are distinct 
in some nuclei, but most nuclei only contain coarse chromatin 
granules scattered uniformly. There is no sign of mitosis, but there 
is a good deal of vacuolation in the cytoplasm in some places, although 
not the characteristic vacuolation caused by thyroid feeding. The 
islets of Langerhans appear normal. 


(c) Effects of Thyroidectomy. 


No. 1 rat of C group. Weight 190 grm. Killed thirty-four days after 
thyroidectomy. 


Studies on the Endocrine Glands 265 


The base of the right lung seems slightly congested. The intestine and 
its contents and the organs of the body generally appear normal. 


Pancreas.—The nuclei of the alveolar cells vary in size from 374 
to 5u, but some are as large as 7'5u, and there are even a few as 
large as 10u. They lie near the base of the cells. Some of them 
contain large nucleoli, with a tendency for the chromatin granules 
to be arranged near the periphery of the nucleus, whilst others have 
uniformly scattered coarse chromatin granules. All the nucleoli are 
stained blue by Mallory. Mixed with the ordinary larger rounded- 
angular cells are many smaller cells. The zymogen granules are 
much fewer than either in the control or in the parathyroid- 
ectomised animal. The cells of the islets and the centro-acinar cells 
exhibit no differences from the normal. 


Summary of the Effects of Parathyroidectomy and 
Thyroidectomy upon the Pancreas. 


1. Parathyroidectomy produces comparatively little change in the 
structural appearances of the pancreas. But the alveolar cells are some- 
what smaller, and their cytoplasm less compact in structure, exhibiting 
numerous small vacuoles and crevices (appearance of canalisation). There 
is rather less zymogen than in the normal pancreas. 

2. Thyroidectomy is followed by the appearance of many small cells 
in the alveoli, and by an enlargement of many of the cell-nuclei. There is 
a greater diminution of zymogen than after parathyroidectomy. 

3. The islets of Langerhans appear unaltered as the result of these 
operations. 

4. No mitoses are visible after either operation. 


Effects of Thyroid Feeding on Pancreas. 


The remaining rats of each of the three groups A, B, and C were, from 
now on, fed with the paste of ground rusks and water (without meat), to 
which was added 1 grm. of dry ox-thyroid per rat per diem: this diet was 
maintained for one week, from March 25 to 31. From the third day of the 
addition of thyroid to the diet the animals of all the groups were observed 
to be less active than before and to be losing weight. The appetite decreased 
to a remarkable extent. Some of the animals showed marked emaciation ; 
several suffered from diarrhoea; and the hair was shed to a much greater 
extent than usual. At the end of the week one rat (No. 2) of each group 
was killed. The following are the findings :-— 


(a) In Unoperated Animal. 


No. 2 rat of A group. Thyroid-fed. Weight 140 grm. 
Both the small and large intestines are somewhat congested. The 
contents of the large intestine are soft. 


266 


Kojima 


Pancreas (figs. 7, 8, and 9, and Plate I, B)—The pancreas is 
pink in colour. The alveolar cells vary considerably in size, but 
the alveoli are, on the whole, smaller than in the animals killed prior 
to the addition of thyroid to the diet. The nuclei, on the other hand, 
are larger; most of them measure from 7y to 10u. The largest con- 
tain coarse chromatin granules uniformly seattered within them, and 
usually stain rather more darkly with hematoxylin than the rest. 
Sometimes there is an enlarged nucleolus. Many of the alveolar 
cells exhibit mitotic figures in various stages of karyo- 
kinesis (fig. 8 and Plate I., B). These dividing cells are large: their 


Fic. 7,—Section of pancreas of rat (male) fed with an addition to the ordinary 
diet of 1 grm, of dry ox-thyroid per diem during seven days. Microphotograph ; 
magnified 90 diameters. Hematoxylin preparation. 

If this is compared with fig. 1, it will be seen that the gland has a more 
compact appearance, and that the inner (zymogen) zone of the alveoli is smaller 
than in the normal animal, Two islets are included in the field. 


cytoplasm is but little stained with hematoxylin, and shows no sharp 
distinction between outer and inner zones. Paranuclei are some- 
times observable in the cytoplasm near the nucleus. The cytoplasm 
of the dividing cells is stained faintly blue by Mallory, whilst the 
scanty and scattered zymogen granules and the paranuclei are stained 
red. So numerous are the mitoses that in places a dozen or more are 
visible within one field of the ordinary high power (600 diameters) of 
the microscope. 

The cells which exhibit mitosis are usually sharply outlined and 
partially separated from the adjacent cells. They usually have a 
spheroidal form. 

Many of the cells which are not undergoing mitosis are also large. 
Their cytoplasm is but little stained with hematoxylin; their nuclei, 


Studies on the Endocrine Glands 267 


which are Jarger than normal, are deeply stained by it but without 
showing distinct chromatin granules. ‘The zymogen granules of the 
alveolar cells are in general scanty as compared with those of the 
animals killed without thyroid feeding (fig. 9; compare with fig. 4). 
Besides the ordinary and the dividing cells, many small cells occur 
packed closely together; they contain small, rather deeply stained 
nuclei, which are often in pairs, and suggest the idea that they are the 
result of division, The small cells contain but few zymogen granules, 
and these are scattered in the cytoplasm of the inner zone. As in 
the normal pancreas, the zymogen is more conspicuous in the alveoli 


Fic. 8.—Portion of a section from the same pancreas as that shown in fig. 7, but 
magnified 500 diameters. Haematoxylin preparation. 

Notice the great variation in size of the alveolar cells, and also.a considerable 
variation in the size of their nuclei as compared with the normal pancreas 
(fig. 2). The coarseness of the chromatin granules within the nuclei is also to 
be observed, and in some a very distinct, centrally situated nucleolus. Several 
of the cells show mitoses. The amount of zymogen in the alveoli is variable, 
but there is much less than in the normal pancreas. In this section the inner 
zymogen-containing zone has remained completely unstained by the hematoxylin. 


which immediately surround the islets of Langerhans. In nearly all 
parts some of the alveolar cells exhibit vacuoles, the vacuolation being 
occasionally marked (fig. 12); it is, however, not uniformly dis- 
tributed. Such vacuolisation is very characteristic of the pancreas 
of the thyroid-fed animal. ‘The vacuoles vary in size; some are large 
enough to occupy a large portion of the cytoplasm, others are smaller. 
They are always quite clear and sharply outlined, and are not to be 
mistaken for the more faintly outlined vacuoles which are seen after 
parathyroidectomy and sometimes in the pancreas of the normal 
animal. The vacuolisation was most marked when fresh sheep- 
thyroid had been used (p. 275). 


268 Kojima 


wet 


ote 


Fic, 9.—Section from the same pancreas as that shown in fig. 8, but stained 
with Mallory instead of hematoxylin. Microphotugraph ; magnified 
100 diameters. 

This photograph is to be compared with that shown in fig. 4. Notice 
the considerable diminution in the amount of zymogen, which is still, 
however, accumulated in greatest amount in the alveoli which are 
adjacent to the islet. 


(b) In Parathyroidectomised Animal. 


No. 2 rat of B group. Thyroid-fed. Weight 205 grm. Killed forty- 
two days after parathyroidectomy. 

Both lungs are slightly congested, especially at the base. The liver is 
also congested. The large intestine is normal in appearance, but its contents 
are soft. The thyroid glands are paler than normal. 


Pancreas.—As seen under a low power, the alveoli appear on the 
whole smaller than in the controls. Both in size and shape the 
alveolar cells vary considerably (as in the unoperated thyroid-fed 
animal). Some cells contain vacuoles. Many of the nuclei of tlie 
alveolar cells are large, measuring from 7'5u to 10u; they are stained 
deeply with hematoxylin, and show a coarse granulation of chromatin, 
with sometimes an enlarged nucleolus. Small nuclei (3°74) belonging 
to small cells are also seen in great number. These are closely set, 
as if they had just completed division. Many of the alveolar cells 
exhibit mitoses. but these do not appear so numerously as in the 
animal just described. Zymogen granules are more scanty than 
normal. The nucleoli, when visible, are, in Mallory preparations, 
sometimes blue, sometimes red. Paranuclei are occasionally seen in 
the cells which exhibit mitosis. As in the case just described, zymogen 
granules are scarce in these dividing cells. Zymogen granules are 


Studies on the Endocrine Glands 269 


more abundant in the alveoli immediately adjacent to the islets of 
Langerhans than in more remote alveoli. 
The islets themselves show no change from the normal. 


(c) In Thyroidectomised Animal. 
No. 2 rat of C group. Thyroid-fed. Weight 170 grm. Killed forty- 
one days after thyroidectomy. 
The intestines are slightly congested. The contents of the large 
intestine are soft. The pancreas is of a pinkish colour. 


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Fic. 10.—Section of pancreas of rat (male) which had been subjected to thyroidectomy 
and subsequently fed with an addition to its ordinary diet of 1 grm. of dry ox- 
thyroid per diem during seven days. Microphotograph ; magnified 90 diameters. 

Great variation is noticeable in the size of the alveoli, largely due to differences 
in the amount of zymogen within the cells. 


Pancreas (figs. 10, 11).—The alveolar cells show a good deal of 
difference in appearance (fig. 10). Their nuclei vary in size from 4 
to 10u. The smaller nuclei are placed closely together; the cells to 
which these small nuclei belong are smaller than the rest. The larger 
nuclei contain coarsely granular uniformly scattered chromatin, but 
some have a large central nucleolus with a tendency for the chromatin 
granules to be arranged near the periphery of the nucleus. A greater 
number of mitotic figures (fig. 11) are observable than in the sections 
of pancreas of the animal belonging to B group; perhaps as many as 
in the unoperated animal, but this is a point not easy to determine. 
Some of the cells show characteristic vacuolation. Zymogen granules 
are in general scanty, although, as usual, in greater number in the 
immediate neighbourhood of the islets. The latter show, as before, 
no special departure from the normal. 


270 Kojima 


On account of the mortality amongst the group of thyroidectomised 
rats fed with thyroid, and with the view of confirming the results 
obtained under these circumstances, another full-grown male rat (No. 120) 
was subjected to thyroidectomy on September 16, 1916. From October 
4. to 9 there was added to its normal diet 1 grm. of dry ox-thyroid per diem. 
From the third day of this diet the animal suffered from slight diarrhcea, 
and appeared inactive, with a distinct decrease of appetite. The skin also 
had an unhealthy appearance. Before the thyroid feeding the weight of 


Fic. 11.—Part of a section of the same pancreas as that shown in fig. 10, magnified 
500 diameters. Hematoxylin preparation. 

Considerable variation in size of the alveolar cells and of their nuclei is apparent. 

The large nuclei contain abundance of coarse chromatin granules which are deeply 

stained with hematoxylin. Several cells show mitoses. There is a general diminu- 
tion in the amount of zymogen (clear zone), 


the animal was 265 grm. After six days’ thyroid feeding it was 250 grm. 
It was then killed. 

Post-mortem.—The contents of the large intestine are soft. The 
weight of the pancreas is 16 grm.; the ratio of the pancreas to body- 
weight is 1:1562. (This will be afterwards referred to.) 


Pancreas.—The alveoli generally are small. Their cells vary in 
size, and also their nuclei; the latter measure between 3°74 and 10x, 
but most are from 5u to 62u. Some of the larger nuclei stain par- 
ticularly deeply with hematoxylin. These have scattered chromatin 
granules. Others contain a large nucleolus, staining red with Mallory, 
with a tendency for the chromatin granules to be arranged near the 
periphery of the nucleus. <A considerable number of cells exhibit 
mitotic figures. These cells are generally of large size. Paranuclet 
which stain red with Mallory are seen in them. Zymogen granules 


Studies on the Endocrine Glands 271 


are singularly scanty throughout. Many cells show vacuoles in their 
cytoplasm. There appears to be no change in the islets. 

Later, another thyroidectomised rat in this series of experiments was 
dieted with thyroid addition during six days, and then killed and the pan- 
creas examined. The cells showed changes similar to those above detailed, 
although the mitoses were relatively fewer in number. In this case also 
the weight of the animal diminished as the result of the thyroid feeding. 


The weights of the respective groups at different periods are shown 
in Tables IL. II., and III. 
TaBe I. 
A Group (Control). 


Date No. of | Weight 
1916. Food. rats. in Se. 
Mar. 11-17 | Lean meat and rusks 5 1040 
» 18-24 se 5 1050 Rat weighing 255 grm., killed 
March 25. 
», 25-31 | 1 grm. dried ox-thy- 4 795 Rat weighing 140 grm., killed 
roid per diem, and (on 25th) April 1. 
rusks 728 
(on Ist) 
Apr. 1-7 | 01 grm. dried ox- 3 588 Rat weighing 140 grm., died 
parathyroid per (on 1st) April 1. Another died on 
diem, and rusks April3. Weight of remain- 


| ing rat (on 7th) 165 grm. 


TaBLeE II. 
B Group (Parathyroidectomised). 


Date No. of | Weight 
1916. Food. rats. in grm. 
Mar. 11-17 | Lean meat and rusks 5 1025 
» 18-24 - 5 1040 Rat weighing 295 grm., killed 
on 25th. 
»» 25-31 | | grm. dried ox-thy- 4 745 Rat weighing 205 grm., killed 
roid per diem, and (on 25th) on April 1. 
rusks 650 
(on 31st) 
Apr. 1-7 | 01 grm. dried ox- 3 445 
parathyroid — per (on Ist) 
diem, and rusks 440°7 
(on 7th) 


| 
VOL. XI., NOS. 3 AND 4,—1917. 18 


272 Kojima 


TaBLE III. 
C Group (Thyroidectomised). 


Date, No. of Weight 
1916. Food. rats. in erm, 
Mar. 11-17 | Lean meat and rusks 4-3 696 One rat died on March 13. 
(end) 
» 18-24 - 3 635 Rat weighing 190 grm., killed 
(end) on March 25. 
», 25-31 | 1 grm. dried ox-thy- 2 445 Rat weighing 170 grm., killed 
roid per diem, and (25th) on April 1. 
rusks | 426 
(31st) 
Apr. 1-7] 01 grm. dried ox- 1 256 
parathyroid per (Apr. 1) 
diem, and rusks 235 
(Apr. 7) 


In connexion with these tables it is to be noted that variations in the 
amount of food within the stomach (which always contains food in the 
rat) cause large differences in the body-weight, the stomach of the rat being 
large in proportion to the size of its body. Care was taken, however, to 
weigh all the animals at the same time of day, viz. between 3 and 4 p.m. 
As is seen in the tables, the weights of the rats belonging to the various. 
groups did not undergo much change during the first fortnight, when the 
animals were fed with a diet composed of rusks and lean meat. But after 
seven days’ thyroid feeding all the weights showed greater or less decrement, 
doubtless owing to the diminished appetite. The loss of weight is least 
in the thyroidectomised rats; greatest in the parathyroidectomised. 

Since two of the rats which had been fed with thyroid and were in- 
tended for control purposes died during the parathyroid feeding (probably 
as a result of the previous thyroid diet), there was only one thyroidectomised 
animal to compare with three parathyroidectomised ones in this period of 
the experiment. 

It appears from the tables that parathyroid feeding made very little 
difference to the weights of the parathyroidectomised rats (B group). 


Effects of Parathyroid Feeding on Pancreas. 


The remaining animals of each group (A, B, and C) were fed as before 
with rusks, but in place of the addition of thyroid material, 0-1 grm. of dry 
ox-parathyroid per rat per diem was added to the diet. This feeding 
lasted for a week (April 1-7). During this third period the appetite 
of all the animals gradually improved. But one rat of the A group and 
two of the B group continued to suffer from diarrhoea for two or three 


Studies on the Endocrine Glands 273 


days after the thyroid feeding had been stopped, and one animal of A group 
died on April 1, i. after seven days’ thyroid feeding, apparently from 
diarrhcea. Its weight was only 140 grm. the day before death, although 
at the commencement of the thyroid feeding it was 165 grm. In this 
animal the large intestine was greatly congested and the contents were 
watery. A second rat belonging to A group died on April 3, ie. after 
thyroid feeding had been replaced during two days by parathyroid, so that 
only one rat of A group was left for further observations. 

At the end of the week of parathyroid feeding one rat (No. 3) of each 
group was sacrificed and the pancreas examined ' 


(a) In Unoperated Animal. 


No. 3 rat of group A. Parathyroid-fed (after thyroid). Weight 
165 grm. Unoperated. Nothing noticeable post-mortem. 


Pancreas.—In most parts the alveoli are about equal in size to 
those of No. 1 animal of A group (normal). There are a certain 
number of small cells, but most are fairly uniform in size. ‘The cell- 
nuclei vary in size from 3°74 to 94; the majority are between 54 and 
62u. Many contain a distinct nucleolus; others merely coarse 
chromatin granules. Both nuclei and nucleoli are stained blue by 
Mallory. No mitotic figures are visible, nor are any vacuoles observ- 
able in the cytoplasm. Zymogen granules are plentiful: many more 
than immediately after thyroid feeding. They are, as usual, specially 
abundant in the alveoli adjacent to the islets. The islets show no 
change. 


(b) In Parathyroidectomised Rat. 


No. 3 rat of group B. Parathyroidectomised forty-nine days before 
death. Parathyroid-fed (after thyroid). Weight 206 grm. Nothing 
noticeable post-mortem. 


Pancreas.—Although there are many small alveolar cells, most 
are moderately large and nearly equal in size: this also obtains for 
the alveoli. The nuclei vary from 5u to 9u, most being between 
5u and 62u. Nucleoli are distinct in many of the nuclei, and the 
tendency of the chromatin granules to be arranged at the periphery 
of these nuclei is observable: other nuclei have coarse chromatin 
granules without any clear differentiation of nucleoli. No mitoses 
are to be found, and there is not much vacuolation of the cytoplasm. 
Zymogen granules are decidedly more abundant than in the rats 
killed immediately after thyroid feeding. The zymogen is, as usual, 
specially marked in the cells adjacent to the islets. The islets them- 
selves show no change. 

1 Tt must be borne in mind that all these animals, which were now fed with addition 


of parathyroid, had previously received thyroid with their food, and the effects of the 
thyroid feeding may not have entirely passed off. 


bo 
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(c) In Thyroidectomised Rat. 


No. 3 rat of group C.  Parathyroid-fed (after thyroid). Weight 
235 grm. Thyroidectomised forty-eight days before death. Post-mortem 
slight congestion is noticeable in the small intestine. 


Pancreas.—The alveoli are distinctly larger than in the rats 
killed immediately after thyroid feeding. Their cells are of nearly 
equal size; they contain abundance of zymogen granules. The 
nuclei vary in size from 3‘7y to 6°24. Many have coarse chromatin 
granules; some an enlarged nucleolus which stains red with Mallory. 
There is nowhere evidence of mitosis, and no apparent change in the 
islets. 


Summary of the Effects of Thyroid and Parathyroid Feeding 
upon the Rat’s Pancreas. 


(1) After a week upon thyroid diet the pancreas of the rat shows 
typical changes, viz. (a) the alveoli are smaller than normal; (b) most 
of the alveolar cells are smaller than normal; (c) the nuclei vary in size, 
many being enlarged and deeply stained; (d) the cytoplasm of the alveolar 
cells exhibits in places a marked and characteristic vacuolation; (e) many 
of the alveolar cells show typical mitotic figures indicative of rapid multi- 
plication; and (f) they contain comparatively little zymogen. 

(2) After intermission of thyroid feeding the pancreas shows a decreas- 
ing number of mitotic figures, whilst the amount of zymogen and the 
general size of the cells again becomes increased, although the variations 
in size of the alveolar cells and of the nuclei are still manifest; the 
alveoli themselves continue for some time to be smaller than normal. 

(3) Addition of a small amount of ox-parathyroid to the daily diet is 
not productive of any marked changes in the pancreas. 

(4) In none of the experiments have any alterations been observed in 
the islets of Langerhans. 


Thyroid Feeding in Different Sexes. 


This series of experiments was primarily intended for the investigation 
of the effects, if any, upon general and gaseous metabolism of feeding with 
thyroid and with parathyroid in the two sexes,! but the animals were sub- 
sequently used for histological observation, and the results of the post-mortem 
findings may therefore be given here. The parathyroid used was obtained 
from the ox, and was dried and powdered. The thyroid was sheep’s thyroid 
obtained fresh from the slaughter-house. 

Three full-grown inale and three non-pregnant female rats were used, 
and were divided into two groups. From April 28 to May 11 all were 
fed on ground rusks and water alone. From May 12 to May 18 there 
was added to this diet 0-1 grm. of dry ox-parathyroid per rat per diem. 


1 The observations relating to metabolism are given in a subsequent paper (p. 351 
of this volume). 


Studies on the Endocrine Glands 275 


From May 19 to May 25 the parathyroid was left out, and the diet 
consisted of ground rusks and water alone. Between May 26 and June 5 
the animals received an addition of 3 grm. of fresh sheep’s thyroid per 
rat per diem to the ordinary diet.1_ During the period of parathyroid feed- 
ing none of the animals showed any special symptom, but from the third 
day of the thyroid feeding they had diarrhcea and their appetite dimin- 
ished. There was also an effect produced upon the skin, the hair falling 
out much more than before. On June 6 all the animals were killed and 
their organs placed in 10 per cent. formol solution. 


Fic, 12.—Section of the pancreas of a rat (female) fed with the addition to the ordinary 
diet of 4 grm. of fresh sheep-thyroid per diem during four days. The animal was 
killed after one day’s intermission of the thyroid. Microphotograph ; magnified 
150 diameters. Mallory’s stain. 

Notice the extensive vacuolation of many of the alveolar cells. Two islets 
are included in the section. 


Two of the animais exhibited marked congestion of both the small and 
large intestines, the contents of which were watery. 


Pancreas.—The alveoli as seen in sections are distinctly smaller 
than normal. The alveolar cells are also generally smaller, but 
vary in size. Their nuclei measure from 3°74 to 11:2u, but most 
of them are between 5u and 7-5u. There are many small alveolar 
cells. These have small nuclei staining deeply with hematoxylin 
and giving the appearance of having undergone proliferation. 
Their nucleoli are enlarged and their chromatin granules coarse; 


5 
zymogen granules are but few in number in these small cells. 


1 This is a little less than 1 grm. of dry thyroid, which is equivalent to 3°5 grm. fresh 
thyroid. 


276 Kojima 


There are a certain number of large round cells with a clear, 
non-stained protoplasm and a comparatively small nucleus deeply 
stained by hematoxylin, whilst other enlarged alveolar cells show 
typical mitoses. Such cells are, however, far fewer than in the 
animals with which we were dealing in Series 1, which were fed 
with 1 grm. of dry ox-thyroid. The zymogen is throughout much 
more scanty than normal. Many cells have large vacuoles in their 


Fic. 13.—Section from pancreas of rat (male) fed with an addition to its ordinary diet 
of 4 grm, of fresh sheep-thyroid per diem during four days. Microphotograph ; 
magnified 400 diameters. Haematoxylin preparation. 

Many of the alveolar cells show large vacuoles. The outer zone of the cells is 
relatively thick. It is more deeply stained by hematoxylin than the inner zone. 
The zymogen is considerably diminished in amount, especially in the cells which 
contain large vacuoles. The nuclei are large and very deeply stained. 


cytoplasm (figs. 12, 13), mainly in the inner zone. The islets of 
Langerhans appear to have undergone no change. 


Summary.—There is no distinct difference produced by thyroid feeding 
in the pancreas of the different sexes. All the usual effects of thyroid 
feeding are well seen in this series, in which fresh sheep-thyroid was 
employed in place of the dry ox-thyroid. There are, however, fewer 
mitoses than in the ox-thyroid fed animals, but, on the other hand, more 
pronounced vacuolation of the cells (this appeared most marked in females) 
and rather greater diminution in their zymogen contents. 


Effect of Thyroid Feeding on Body-weight in Different Sexes. 


The following table shows the weights at different periods as well as 
those of control groups :— 


Studies on the Endocrine Glands 277 
TABLE IV. 

| Control 

Date, : | = ., |Control (male| (young male | 

| 1916. A group (male). | B group (female). | 144 female) | “and preg- | 
nant female). 
] 2 3 4 5 6 107 | 108 | 105 | 106 
April 28 155 145 235 180 140 180 180 150 90 148 
May 5 155 | 140 230 180 140 185 182 150 100 148 


11 | 160 | 145 | 230 | 180 | 146 | 190 | 185 | 154 | 100 | 145 
»  12| 160 | 145 | 924 | 175 | 146 | 187 | 185 | 155 | 105 | 140 
18 5 | 140 | 225 | 170 | 145 | 185 | 180 | 155 | 105 | 140 
S619] 166 | °140 | 985 | 173) 147 | 185] 180 | 155 | 105 | 145 
» 25 | 165 | 145 | 287 | 175 | 150 | 190 | 185 | 156 | 110 | 145 
»  26| 165 | 145 | 298 | 177 | 150 | 190 | 185 | 155 | 110 | 145 
June 5 | 140 | 126 | 201 | 160 | 140 | 172 | 186 | 150 | 118 | 152 


~ 
- 
_ 
~ 
oP) 
7 


From May 12 to 18, groups A and B were fed with parathyroid; from May 26 to 
June 5, with fresh sheep-thyroid. 


It would appear from this table that there is no distinct difference in 
the sexes in the effect on body-weight produced by thyroid feeding. 


Feeding with Thyroid for Varying Periods. 

In the following experiments 4 grm. of mixed fresh sheep-thyroid per 
rat per diem was added to the food. 

(a) Male Rat (No.7). Weight 220 grm.—In this animal a single dose 
of the sheep-thyroid was given with the food at 9 a.m. on June 10. Sub- 
sequently it was allowed an unlimited amount of rusk paste. Exactly 
twenty-four hours after taking the thyroid the animal was killed. No 
special symptoms observable. 

(b) Male Rat (No. 8). Weight 230 grm.—Fed with addition of 
4 grm. fresh thyroid to food during two days (June 10 and 11). Killed 
on June 12. No symptoms observable. 

(c) Male Rat (No. 9). Weight 230 grm.—Fed with addition of 
4 grm. fresh sheep-thyroid during three days (June 10 to 12). On 
June 12 slight diarrhcea observable, and the appetite had begun to 
decrease. Killed on June 13. 

(d) Male Rats (Nos. 10, 11, 12,13). Weights 220 grm., 210 grm., 
225 grm., and 105 grm. respectively.—Fed during four days (June 
10 to 18) with addition of sheep-thyroid. 

(e) Male Rats (Nos. 14 and 15). Weights 180 grm. and 210 grm. 
respectively.—These rats were fed from June 10 on a diet containing 
fresh sheep-thyroid. 


No. 10.—Began to have diarrhcea on June 12, and showed diminu- 
tion of appetite. Killed on June 14. It had lost 15 grm. in weight. 
No. 11.—No special symptoms observed. On June 13 it had 
lost 10 grm. in weight. Killed after one day’s intermission of feeding. 


Kojima 


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No. 12.—Suffered from slight diarrhcea from the third day of 
feeding, and appetite somewhat affected. At the end of the feeding 
it had lost 15 grm. in weight. Killed after two days’ intermission 
of feeding. 

No. 13.—Diarrhcea from third day onwards, with decreased 
appetite. Killed three days after cessation of thyroid feeding. 

Nos. 14 and 15.—Animals suffered from diarrhcea from the fourth 
day of feeding. Their appetite also decreased, and they were com- 
paratively inactive. No. 14 was killed after five days, and No. 15 
after six days of the thyroid feeding. Their weights then were 
164 grm. and 193 grm. respectively. 

In all these animals the intestines, both small and large, show 
congestion. There are no other noticeable changes post-mortem. 

Pancreas. Nos. 7 and 8 (one and two days’ thyroid 
feeding).— There is as yet no evidence of karyokinesis. The 
zymogen granules are scanty, but otherwise there is no difference 
from the normal, except that No. 8 (two days’ feeding) already 
begins to show the marked vacuolation of some of the alveolar 
cells which is characteristic of thyroid feeding. 

No. 9 (three days’ thyroid feeding).—Both the alveoli and 
the cells vary more in size than in the normal, and this is also the 
case with the cell-nuclei, which measure from 3u to 9u. A few are 
of still larger size (10u), and many are more deeply stained than 
normal. Some show enlarged nucleoli. Both nuclei and nucleoli 
are stained blue by Mallory. The nuclei have abundant chromatin 
granules. A few cells show typical mitoses. The zymogen granules 
are scanty in all the alveoli. Many of the cells exhibit vacuoles. 

Nos. 10-15 (four to six days’ thyroid feeding).—The alveoli 
are smaller than normal, and contain many small cells. A consider- 
able number of the alveolar cells are in mitosis. These cells are 
relatively large. The mitoses are much more abundant in the rats 
which have been fed for five and six days than in the four-day 
animals. The nuclei of the cells vary considerably in size (3u to 9u). 
Some of the large nuclei contain very distinct nucleoli staining 
deeply with hematoxylin. Both nuclei and nucleoli stain blue by 
Mallory. The zymogen granules are scanty throughout. Many of 
the alveolar cells exhibit vacuoles of varying size. Small vacuoles 
are observable in the islet-cells. 


Summary.—In all the animals of this series (like those animals of the 
last series in which fresh sheep’s thyroid was also given) the usual effects 
of thyroid feeding are observed. The zymogen granules are considerably 
diminished; more so than in animals dieted with dry ox-thyroid. This 
change is seen even after a single dose, whilst vacuolation of the cells is 
visible after two days’ feeding. 

Mitotic figures began to appear after three days’ feeding; their number 
gradually increased up to six days. 

Most of the animals suffered from diarrhcea, and their appetite 
decreased after three days’ feeding; the weights in consequence diminished 
throughout. 


Studies on the Endocrine Glands 279 


Thyroid Feeding with Intermission, and also with an 
Increased Dose. 


In these experiments dry ox-thyroid was added to the usual rusk paste. 
Nine full-grown male rats were used, and were divided into three groups, 
A, B,and C. The experiments were begun on July 19. 

A group. Control—Two animals belonging to this group, viz. rats 
Nos. 16 and 18, were killed on August 30; the third (rat No. 17) was 
kept alive. One animal of the group suffered from slight diarrhoea for a 
few days. 

B group (rats Nos. 19, 20, 21). Thyroid fed, with Intermittence. 
—These animals were fed on the rusk diet alone until August 1. From 
August 2 to 8 they were put on thyroid diet. From August 9 to 15 this 
was stopped, and they returned to the simple rusk diet. From August 
16 to 22 they again received 1 grm. of dry ox-thyroid per rat per diem. 
From August 23 to 29 they had the rusk diet alone. Most of the animals 
suffered from diarrhcea during the first thyroid feeding. Their appetite 
decreased considerably. They took a considerable amount of water. They 
were inactive, and lost in weight: there was more falling out of hair than 
before. During the second thyroid feeding the condition of the animals 
was better, their appetite being less affected. Nos. 19 and 21 were 
killed on August 30, and No. 20 on September 16; i.e. twenty-five days 
after the second thyroid feeding had ceased. 

C group (rats Nos. 22, 23, 24). Thyroid fed, with Increasing 
Dose.—The rats of this group were fed on ordinary rusk paste from July 19 
to August 1. From August 2 to 8 they received 1 grm. of dry ox-thyroid 
per rat per diem. From August 9 to 15 the amount was increased to 1:2 
grm. From August 16 to 22 the amount given was 1:3 grm. From August 
23 to 29 they received 1:5 grm. per rat per diem. 

During the first week of thyroid feeding there was decrease of appetite, 
inactivity, and diarrhoea. During the second the same symptoms were 
seen, and the falling out of the hair was much more marked. During the 
third and fourth periods the general condition of the animals improved. 

Effect on Structure of Pancreas.—aAll these animals were killed 
on August 30. Nos. 16 and 18 showed no appreciable departure from 
normal. In Nos. 19, 20, and 21 the pancreas appeared somewhat enlarged 
as compared with the controls. The colour was light pink, paler in 20 than 
in 19 and 21. 

In Nos. 22, 23, 24 the pancreas was of a pink colour, and appeared 
somewhat enlarged. In No. 24 the intestines were a little congested. 


Pancreas.—Examination of the pancreas of the rats employed in 
Series 4 (thyroid feeding with intermission and with increasing dose) 
gave the following results :—In Nos. 19, 20, 21 (Group B) the alveoli 
generally are large, although quite small in some parts. Many of 
the alveolar cells are large, but they have small ones amongst them; 


280 : Kojima 


the small cells are especially abundant in the small alveoli. Most 
of the nuclei are large; there is, however, considerable variation in 
size (3°74 to ll). The nuclei of the small cells are smaller than 
the rest, and stain more deeply. The nucleoli generally are large, 
and with Mallory are stained red, the nuclei being coloured blue. 
Most contain abundant coarse chromatin granules. Some of the cells 
show mitotic figures, but these are not very numerous. Portions 
of the pancreas, consisting usually of isolated lobules, are here and 
there seen to be wholly formed of small cells which have lost their 
typical alveolar arrangement and exhibit signs of having recently 
undergone proliferation; indeed, some of them still exhibit mitoses! 
(see Plate II.). 

Zymogen granules are far less numerous than in the normal 
pancreas. These granules, which are stained very distinctly by 
Mallory, form a red mass in the ordinary cells, but in the smaller 
cells they are very scanty and are scattered in the cytoplasm, not 
massed together. Vacuoles, some quite large, are present in many of 
the alveolar cells. Vacuolation may also be seen in some of the cells 
of the islets of Langerhans. 

In rats Nos. 22, 28, 24 (group C) the changes in the pancreas 
are similar to those just detailed. The controls (Nos. 16 and 18, 
group A) show normal appearances. 


Summary.—After thyroid feeding with intermission, and also if the 
dose of the thyroid be increased, the pancreas is larger in proportion to the 
body-weight than in the controls. It shows, besides the ordinary alveoli, 
others which consist entirely of small cells not regularly arranged into 
alveoli and containing very little zymogen. A certain number of the 
alveolar cells show mitoses, and many of them have large vacuoles. 

A second thyroid feeding following on the first after a certain interval 
appears to produce less effect than the first. 


Effect of Thyroid Feeding on the Weight of the Pancreas. 


The weight of the pancreas was examined in the three groups which 
have just been noticed (Table VIII.). In order to determine the weight 
the pancreas was entirely freed from mesentery with the assistance of 
a lens, and was weighed in all cases at as nearly as possible the same 
degree of moisture. In order to compare with the weights obtained in 
these experiments, the pancreas of nine other full-grown normal male 
rats were also weighed (see Table IX.).2 These tables show a distinct 
increase of pancreas-weight relatively to body-weight in the thyroid-fed 
animals. 

The increase in size of the pancreas which is produced by the thyroid 
feeding is illustrated in the photographs shown in fig. 14. 


1 Mitoses are seen in the rat’s pancreas as long as three days after thyroid feeding has 
been stopped. 


* The weight of the pancreas was not taken in any of the early experiments. 


Studies on the Endocrine Glands 281 


Effect of Thyroid Feeding on the Amount and Quality 

of the Urine. 

The animals took a considerable amount of water during the periods of 
thyroid feeding, so that the amount of urine was thus increased ; but when 
the feeding was intermittent it was not much diminished in the intervals. 
No trace of sugar or albumen was at any time found in the urine of the 
thyroid-fed animals. The most striking effect upon the urine is 
that during and for some time after thyroid feeding iodine 
is always present, whereas in the normal animal fed on rusks alone 
iodine can never be detected in the urine. One rat (No. 20, B group) 


Fic, 14.—Photographs (natural size) of the paucreas of three rats showing the effect 
upon the general appearance of the gland of thyroid feeding. (1) Pancreas from 
rat No, 19 ; (2) pancreas from rat No, 20; (8) pancreas from rat No. 17 (control). 

Both No. 19 and No, 20 received thyroid in addition to their ordinary food 
during two periods with a few days’ intermission. No, 19 was killed a week 
after the second period had ceased, and No. 20 twenty-five days afterwards. 
In all the figures the splenic end of the pancreas is placed uppermost. Both 
(1) and (2) are larger and heavier than (3). 


which was killed twenty-five days after thyroid feeding had ceased still 
showed slight evidence of iodine in the urine. 


The following method was employed!:—10 e.c. of the urine to 
be examined is evaporated to dryness on the sand-bath, and about 
twice its weight of pure NaOH, prepared from metallic sodium, is 
added to the residue; the mixture is heated gradually: at short 
intervals potassium nitrate is added. The heating is continued until 
the mass, at first charred, just becomes white. After being allowed 
to cool, it is extracted with hot water. The extract is filtered into 
a test tube, cooled, acidified with sulphuric acid, and an equal quantity 
of chloroform added. On shaking up the mixture, any iodine 


1 This method is described in Milroy’s Practical Physiological Chemistry, 1904, p. 3. 


282 Kojima 


present is taken up by the chloroform, which now forms a rose-red 
stratum at the bottom of the test tube. 


Tables V., VI., and VII. show the body-weights, food consumed, quantity 
of urine, ete., in these groups. 


TABLE V. 
A group (Control). 
Weight in grm. : 
Gicuaey Amount of food Quantity Specific 
Date, S __| (tusks) con- of urine gravity Reaction 
> = fo} } 
WIG. | Nei] No. | Wo. | ENCE PE PEE eer | oi crimes cae 
16. Vi: 18. PN a os 
July 19-25 300 | 270 | 251 330 246 1:060 | Alkaline 
July 26- 296 | 276 | 245 265°7 250 1-062 _ 
Aug. 1 
Aug. 2-8 281 | 270 | 250 264°3 274 1-060 £ 
» 9-15 | 296 | 272 | 256 281 270 1:058 re 
, 16-22 | 285 | 276 | 258 280°8 | 250 1-059 ke 
» 23-29 | 284 | 276 | 254 275°4 270 1-060 - 
TaBLE VI, 
B group (Thyroid Feeding with Intermission). 
Weight in grm. : 
Genes Amount of food | Quantity Specific 
Date, (rusks) con- of urine gravity Reaction 
1916. No. | No. | Ne. eat Deer ad ee Graeeel of urine. 
1034) 20a | ei Pena m 
July 19-25 185 315 192 380°8 362 1-061 Alkaline 
July 26- 182 310 185 36)) 370 1-060 s 
Aug. 1 
Aug. 2-8} 154 292 161 117 250 1:062 5 
1 grm. dried ox- 
thyroid per 
rat per diem 
eo =15> |) al66 296 175 187 300 1:060 +s 
16-9924) 161 | 291 | 176 | 230:2 | 325 1-061 28 
1 grm. dried ox- 
| thyroid _ per 
rat per diem 
, 23-2941 165 | 309 | 185 | 250 310 1060 | aan 
1 From August 4 the appetite decreased and there was inactivity, with diarrhea. 
2 The appetite is slightly increased. Some of the animals have diarrhea. 
3 The appetite is increased. There is slight diarrhea. 
4 


The general condition is improving. 


Studies on the Endocrine Glands 283 
TaBLE VII. 
C group (Thyroid Feeding with Increased Dose). 
. jatebe cig Amount of food | Quantity | Baanie'| 
Date, S di | (rusks) con- of urine | ee oge Reaction 
1916. No. | No. | No. |Stumed per week| per week | OF ing, | of urine. 
29. 233. 94 in grm. in cc, 
July 19-25 | 175 | 222 | 206 380°3 312 1060 | Alkaline 
July 26- 176 | 218 | 207 360°7 340 1-062 of 
Aug. 1 
| 
Aug. 2-8! 163 192 186 145°2 290 1062 a 
1 grm. dried ox- 
thyroid per 
rat per diem 
» 9-152 | 165 | 191 | 184 155 310 1:060 ra 
1:2 grm. dried 
ox - thyroid | 
per rat per 
diem 
», 16-22% | 164 193 185 180 300 1:060 3 
1:3 grm. dried 
ox - thyroid 
per rat per 
diem 
» 23-29! | 171 | 192 | 189 190 295 1:060 fi 
15 grm. dried 
ox - thyroid 
er rat per 
iem 
1 From August 4 the appetite decreased and there was inactivity, with diarrhea, 
2 As last week, but with much more falling out of hair. 
3 The general condition is better. ‘ Improvement maintained. 
Taste VIII. 
WrIGHT OF PANCREAS AND ITS Proportion 10 Bopy-WEIGHT IN THE NINE 
ANIMALS USED IN THIS EXPERIMENT. 
A group (control). B group. C group. 
No Nol) Nave aNon WoINo. No. | Noi) No, | No; 
16. ie 18. 19. 20, 21. 22. 23. 24, 
Body-weight in grm. | 284 | 276 | 254 | 165 | 309 | 185 | 171 | 192 | 189 
Weight of pancreas} 13 12 Mel 19 1-2 11 12 | 
in grm. 
Proportion of weight |1 :218 Pe 2b2 it 1506S |: W541 7155 ]1 ; 160.1 3171 
of pancreas to body- 
weight in grm. 


284 Kojima 


TABLE IX. 


WerIGHT OF PANCREAS AND ITS PRoporTION TO Bopy-WEIGHT IN NINE 
Norma ApuLT Mate Rats. 


Body-weight in grm. | 145 | 145 | 165 | 185 | 185 | 190 | 195 | 195 | 250 


Weight of pancreas; ‘74 64 65 8 9 | 0°84 OF 1) 1EO8™ | MEZS 
in grm. 


Proportion of weight |1 : 196 |1 : 227 |1 : 254 /1 : 234 |1 : 206 |1 : 226 |1: 215 |1L:181|1 2195 
of pancreas to body- 
weight in grm. | 


Effects of Smaller Doses of Thyroid. 


To determine whether smaller doses of thyroid than those we were 
accustomed to use would produce the effects described, the amount of 


Fic. 15.—Pancreas of rat (male) which had been fed with an addition of 0°5 grm, of 
dry ox-thyroid per diem to the ordinary diet during seven days, i.e. half the 
quantity which had been previously used. Microphotograph ; magnified 450 
diameters. Hematoxylin preparation. 

Notice again the variation in size of the alveoli as well as of their cells: also 
the occurrence of very large nuclei which contain abundance of coarse chromatin 
granules staining deeply with hematoxylin. Several of the celis exhibit mitosis. 


thyroid in the food was reduced to one-half, viz. to 0°5 grm. of dry ox- 
thyroid per rat per diem administered with the rusk paste. Two full- 
grown male rats (Nos. 25 and 26) were put on this diet from July 19 
until July 25. Neither of these animals suffered from diarrhcea, but 
their appetite and weight decreased from the fifth day of the feeding 
onwards. The weights were respectively 180 grm. and 185 grm. at the 
beginning of the feeding, and 175 grm. and 170 grm. at the end. The iodine 


Studies on the Endocrine Glands 285 


test in the urine gave a positive reaction from the second day of feeding 
onwards. At the termination of the period the animals were killed. 


Pancreas.—The alveoli are somewhat smaller than normal, and 
although most of the alveolar cells are fairly equal in size, a consider- 
able number of small cells are included amongst them (fig. 15). The 
nuclei vary in diameter from 3°74 to 7°54; a few larger ones are seen 
(104). These contain abundance of chromatin granules. Zymogen is 
but scanty. Many cells exhibit mitosis, and a few show vacuolation. 


Summary.—These experiments make it evident that a much smaller 
amount of added thyroid than that used in most of our experiments is 
sufficient to cause the effects on the pancreas which have been described as 
characteristic of thyroid feeding. 


Effects of Prolonged Feeding with Thyroid. 


The effects of prolonged feeding were next investigated : accordingly, the 
following experiments extended over a month. The material used. was 
partly dry ox-thyroid, partly fresh sheep-thyroid. Four full-grown female 
rats (non-pregnant) were separated into two groups, A and B, and fed with 
rusk paste. To the food of A group 1 grm. of dry ox-thyroid per rat per 
diem was added: to that of B group 4 grm. of fresh sheep-thyroid. The 
experiment lasted from May 28 until June 26. From the fourth day of the 
feeding one of the two rats of A group had marked diarrhcea, and all the 
other animals of both groups suffered from slight diarrhoea. The appetite 
in all was diminished, but after the second week gradually recovered. By 
the end of the fourth week the animals were all healthy and showed no 
special symptoms. ‘They had also increased in weight, although they lost 
in weight during the first period of feeding. They were killed on June 27. 
The weights at different periods are given in Table X. 


TABLE X. 
A group. B group. 

Date, 

1916 
No. 27. No. 28. No. 29. No. 30. 
Body-weight in grm. | May 28 195 180 173 185 
. June 4 172 165 156 172 
3 fe atl 170 170 159 170 
a Hits 180 170 163 175 
= oy PAD 184 180 165 189 


Pancreas.—All the rats in this series of experiments show much 
the same appearances in the pancreas. The alveoli are for the most 


286 Kojima 


part large and of nearly equal size. The cells are also large, although 


there are a fair number of smaller cells amongst them. The cell- 


nuclei vary in diameter from 3-7 to 9u; a few of them are as large 
as 10u. Both nuclei and nucleoli stain blue by Mallory and deeply 
by hematoxylin. The chromatin granules of the nuclei are coarse 
and abundant. Many of the cells contain vacuoles of various size. 
These are most marked in the animals which were fed with fresh 
sheep-thyroid. The vacuoles, as is generally the case, are not 
uniformly distributed, but are confined to particular portions of the 
pancreas. Zymogen granules are plentiful in all the alveolar cells. 
There is no evidence of mitosis, and no apparent change in the islets 
of Langerhans. 

Summary.—After feeding for about a month with the ordinary dose 
of thyroid mitoses are no longer seen in the pancreas. and zymogen 
granules have again become plentiful, but there is still considerable 
vacuolation of cells, especially in animals which were fed with addition 


of fresh gland. 


Is the Active Substance or Autacoid extracted by Water, 
and is its Activity abolished by Boiling? 


The next experiments were designed to determine whether (a) extract 
of thyroid made with water and (b) the residue which is left after such 
extraction produce the same effects as feeding with the whole thyroid, 
and further to determine whether boiling destroys the active substance. 

Two sets of observations were made: one with addition of sheep-thyroid, 
the other with ox-thyroid (see Tables XI. and XII). The rats, eight in 
number, were divided into four groups—A, B, C, and D. To make the 
extracts, fresh glands, after being freed from the surrounding connective and 
adipose tissue, were minced, and left standing for an hour in a definite 
quantity of distilled water. The fluid was then filtered. The filtrate was 
divided into two parts: one part was added to the food of A group with- 
out boiling; the other part was boiled and added to the food of B group. 
The remainder left after extraction with cold water was repeatedly washed 
with fresh water, which was decanted off: the residue was then divided 
into two parts. One part was given unboiled to C group, whilst the 
other part was boiled in a small quantity of water and added to the food 
of D group. 

The amount administered was equivalent to 4 grm. of fresh thyroid 
per rat per diem. 

Sheep-Thyroid.—In the animals of groups A and B which received 
the water-extract of sheep-thyroid no special symptoms were observable 
except a slight decrease of appetite. In the animals which had the residue 
there was slight diarrhcea from the fourth day onwards, as well as 
diminution of appetite. After five days all were killed (on July 26). 
In all the groups the iodine reaction in the urine was positive during 
the feeding. 


Studies on the Endocrine Glands 287 


Ox-Thyroid.—In this case some of the rats of the C and D groups 
had diarrhcea from the third day. The appetite of the animals belonging 
to D group seemed to be affected more than the rest. As with the sheep- 
thyroid, the iodine test was positive in every case. 


TaBLeE XJ.—WeIGHTs or RATS FED WITH WATER-EXTRACT OF SHEEP-THYROID 
AND WITH THE Resrpvur (BorLED AND UNBOILED). 


| | | 
A group. B group. C group. D group. 
Date, ' | 
1916. 


Body-weight in grm.| July 20; 170 | 180 | 180 | 185 | 150 | 155 | 190 | 170 


” ” 


TasLeE XII.—WeicuHts or Rats FED WITH WATER-EXTRACT OF Ox-THYROID 
AND WITH THE ReEsIDUE (BOILED AND UNBOILED). 


| 
A group. B group. C group. D group. 
Wyre 2 i ere ee ee 


W916. | wo. | Won aoe |e Wee) oo. | ite. fio: | No: 
| 39, 4O. Al. 42. | 43. 44, 45, 46, 


Body-weight in grm.| July 26| 105 | 100 | 150 | 165 | 240 | 280 | 215 | 180 


31 | 100 120 150 170 236 270 200 180 


° ” 


All the animals were killed on July 29, after five days’ thyroid feeding, 
Post-mortem there was nothing to remark except that the intestines 
were slightly congested in two of the animals (Nos. 45 and 46). 


Pancreas.—On microscopic examination of the pancreas of the 
rats belonging to the A groups (Nos. 31, 32, 39, 40) a great varia- 
tion in the size of the alveolar cells is noticeable. Their nuclei 
measure from 3°74 to 75~ most being fairly large. They con- 
tain abundant coarse chromatin granules and comparatively large 
nucleoli, which in some cases (Nos. 39 and 40) are stained red by 
Mallory, in others (Nos. 31 and 32) blue. Zymogen granules are 
scanty. Many of the cells contain small vacuoles. A few exhibit 
mitoses. 

In those belonging to the B groups (Nos. 33, 34, 41, 42) there are 
more small cells mingled with the larger cells of the alveoli than 
in the A groups. Most of the nuclei are large (some were 10,); 
each of these large nuclei contains a large nucleolus and abundant 
coarse chromatin granules. They are stained deeply by hematoxylin 
(especially in Nos. 34 and 41). There are also a fair number of small 

VOL. XI., NOS. 3 AND 4.—19117. 19 


288 Kojima 


nuclei (5) (especially in Nos. 38 and 42). In preparations from all 
these animals mitotic figures are seen here and there in the sections 
of pancreas (fig. 16). Zymogen granules are on the whole scanty, 
but there are individual differences. Some of the alveolar cells have 
vacuoles of medium size. 

In the animals belonging to groups C and D (Nos. 35, 36, 37, 38, 
and 43, 44, 45, 46) the alveoli have much the same appearance as 
in the others, but the outer zone is somewhat broader in proportion 


a 


ae 


Fic. 16.—Section from pancreas of rat (male) fed with an addition to its ordinary 
food of water extract (boiled) of 4 grm. fresh sheep-thyroid per diem during five 
days. Microphotograph; magnified 450 diameters. Hematoxylin preparation. 

Notice again the variations in size of the alveolar cells and the remarkable 
enlargement of their nuclei, which contain abundance of coarse chromatin 
granules staining deeply with hematoxylin. In some of the cells vacuoles can 
be made out, One cell in the field shows mitosis. The amount of zymogen is 
considerably diminished as compared with the normal. There is no apparent 
change in the islet-cells, the nuclei of which are not enlarged and are of nearly 
equal size. 


to the inner, and there are many more small cells. The diameter 
of the nucleus varies between 3°7» and 7-5. A few are as large 
as 9u or 10u. The nuclei are stained deeply by hematoxylin; they 
contain abundance of coarse chromatin granules. The nucleoli are 
somewhat enlarged, and are for the most part stained red by Mallory. 
Mitoses are here and there present. The amount of zymogen varies, 
being plentiful in some parts and scanty in others; the diminution 
is especially marked in the smaller cells. Vacuolation is visible in 


many of the alveolar cells, and to some extent in the cells of 
the islets. 


Summary.—The mitotic and other changes which have been already 
described in the pancreas from administration of fresh and dry thyroid are 


Studies on the Endocrine Glands 289 


seen also to obtain with water-extracts of the gland, both boiled and 
unboiled ; and with the residue, both boiled and unboiled, after extraction 
with water. 


Does Alcohol-Extract or Ether-Extract of Thyroid contain 
the Autacoid which affects the Pancreas? 


Experiments were next devised to determine whether alcohol- and ether- 
extracts of thyroid have similar effects to water-extracts. The method 
of preparation was as follows:—5 grm. of finely ground completely dry 
ox-thyroid was extracted with absolute alcohol for twenty-four hours, the 
extract being then filtered. The residue on the filter was again thoroughly 
washed with absolute alcohol. This residue was completely dried at 37° C., 
and was then extracted with pure ether for another twenty-four ‘hours. 
The ether-extract was filtered, and the residue on the filter thoroughly 
washed with pure ether. This second residue was also dried at 37°C. 
The alcohol- and ether-extracts were respectively evaporated to dryness. 
In each case, after evaporation of the alcohol and ether, the residue which 
remained at the bottom of the beaker was in the form of a soft paste. 
That from the alecohol-extract will be termed A; that from the ether 
extract, B; and the final residue after successive extraction with alcohol 
and ether will be termed C. Two rats (group A, Nos. 126, 127) were fed 
with an addition of A to their food. Two rats (group B, Nos. 128, 129) 
had an addition of B to their food. Lastly, two rats (group C, Nos. 130, 
131) received an addition of C. Each addition represented the alcohol-, 
ether,- and water-extract respectively of 1 grm. of dry thyroid. 

The groups were fed with the above additions during five days 
(October 26-30). After four days’ feeding the appetite of the animals 
belonging to group C appeared to be affected, but nothing was noticeable 
in the others. All were killed on October 31. There was apparently no 
change in the weight of any of the groups. The iodine test reacted 
positively in the urine of group C; a trace of iodine was observable in 
the urine of group A, but the test was entirely negative for the urine 
of group B. 

Post-mortem nothing particular was observable. 


Pancreas.— On microscopic examination of the pancreas no 
departure whatever from the normal is to be seen in the rats to 
which the alcohol- and -ether-extracts of thyroid were given, but the 
pancreas of the animals which were fed with the residue after such 
extraction with alcohol and ether shows changes similar to those 
obtained in all other cases of thyroid feeding; these changes include 
the appearance of a large number of cells which exhibit mitoses, 
and a general diminution of zymogen granules in the cytoplasm of 
all the alveolar cells. 


Summary. — Alcohol- and ether-extracts of thyroid produce no 
appreciable changes in the pancreas. The residue, after extraction with 


290 Kojima 


alcohol and ether, has, when administered with the food, an effect similar 
to that produced by whole thyroid. 


Effect of Subcutaneous Injection of Thyroid Decoction. 


The effects of subcutaneous injection of a decoction made from dry ox- 
thyroid by boiling with distilled water was investigated in a male rat 
(No. 55), weight 150 grm., the equivalent of 1 grm. of dry ox-thyroid being 
administered per diem for five days. The animal showed no special 
symptoms with regard to appetite, etc. At the end of the period the 
weight was 130 grm. From seven hours after the first injection onwards 
the test for iodine in the urine was positive. 

Post-mortem there is very little to record. The stomach contained only 
a small amount of food, and the intestines appeared slightly congested. 


Pancreas.—The weight was 0°6 grm. The alveoli are both large 
and small, and this applies also to their cells. The nuclei vary from 
3°7u to 9, a considerable number being large and many deeply 
stained by hematoxylin; there are abundant coarse chromatin 
granules. Mitotic figures are seen in some of the cells. There is 
but little diminution of zymogen. No change is observable in the 
islets. 


Summary.— Subcutaneous injection of thyroid decoction produces 
similar changes in the pancreas to those produced by thyroid feeding, but 
less marked. Probably the active substance is only in part taken up by 
the water used for extraction. This is, indeed, shown by the fact that in 
the feeding experiments the residue after extraction by water produces 
all the usual effects. 


DorEs CASTRATION AFFECT THE RESULTS OF PROLONGED 
THYROID FEEDING 7 


The effect of thyroid feeding upon the pancreas of castrated male rats 
was next investigated. But it was first necessary to determine whether 
changes are produced in the pancreas by castration alone. 

Four full-grown male rats were castrated on May 16, and all were kept 
in one metabolism cage, four other entire male rats of similar size being 
kept in another cage as a control. All eight were fed on the standard food 
of rusks and water. During the first week after the operation the appetite 
of the castrated animals was somewhat less than that of the others, but it 
gradually improved ; after a few days they took considerably more food than 
the controls, and their weights increased to a remarkable extent (Table 
XIII.). This increase appears to have been largely due to an addition to the 
adipose tissue, especially that under the skin and in the mesenteric folds. 


Studies on the Endocrine Glands 291 


Tasite XIII.—Errects or CASTRATION ON WEIGHT OF Rats. 


Castrated. Control. 


No. No. | No. No. No, No. | No. | No. 
110. Vath bee lelic: ol Lid, | LIS. | 116; | 17 


Body-weightingrm.| May 16 | 245 | 275 | 275 | 285 250 | 260 | 250 | 265 


| June 9} 245 | 290 300 300 260 260 255 250 


| July 15} 240 | 300 | 3151] 3101| 260 | 265 | 250 | 250 


2 Pn oil AB | BOOS I tees oA 255 | 2651| 255 | 250! 
2 a 0 nee (es 2d tae 2 260 rks 255 


1 Killed on this date. 


Does Castration alone affect the Pancreas? 


Of the castrated rats two (Nos. 112 and 113) were killed thirty days 
after the operation, and the two others (Nos. 110 and 111) forty-nine and 
fifty-seven days respectively. 

Post-mortem the only obvious appearance was the increase of fat 
which has just been mentioned. 

Pancreas.—On microscopic examination of the pancreas the 
alveolar cells appear of nearly equal size throughout, but rather 
smaller than in the normal gland. Their nuclei vary between 3°74 
and 5u. They have fine chromatin granules, and the nucleoli stain 
red with Mallory. Some of the cells show a few small vacuoles in 
their cytoplasm. There is no evidence of mitosis. Zymogen granules 
are in most parts fairly plentiful. 


Does Previous Castration affect the Results of 
Thyroid Feeding on the Pancreas? 


Another lot of four castrated male rats were now taken for the 
experiment of thyroid feeding. These were operated upon on May 25, 
and were separated into two groups, A and A’; four other entire male 
rats of equal size being also taken and separated at the same time into 
two groups, B and BY. To A and B, 3 grm. of fresh sheep-thyroid 
per rat per diem was administered along with the rusk diet, while to 
A’ and B’ an equal amount of fresh lean meat (mutton) was added 
instead of thyroid. 

All the castrated animals (A and A’) were inactive during the first 
week, apparently as the result of the operation. But even those with 
thyroid feeding suffered very little from diarrhcea, and the appetite was 
fairly good. After the first week the thyroid-fed animals remained com- 
paratively inactive in spite of the fact that the wound had completely 


292 Kojima 


healed; there was some loss in weight, but only quite small. On the 
twentieth day after the commencement of thyroid feeding one of the 
animals of group A (No. 47, see Table XIV.) ceased to take food and had 
considerable diarrhcea: it was therefore killed. The condition of the other 
rat of group A was improving, the appetite gradually increasing and the 
loss of weight being but slight. This animal was killed twenty-nine days 
after the commencement of the feeding. The animals belonging to group B 
remained well during the first week of the feeding, but later they gradually 
became less active and their appetite appeared to be affected, the weights 
becoming reduced. From June 16 onwards (twenty-one days) they were 
for the most part lying prone on the floor of the cage. The rats forming 
group B (Nos. 49 and 50) were killed on the twenty-seventh and twenty- 
eighth days respectively (June 21 and 22). 

The rats belonging to groups A’ and B’ remained throughout perfectly 
well. A positive iodine test was obtained throughout the whole experiment 
in the urine of groups A and B (thyroid-fed). In groups A’ and B’ (without 
thyroid) the test always proved negative. 

Those forming group A’ (Nos. 51 and 52) were killed on June 23, and 
one of the rats belonging to group B’ (No. 54) was killed on June 18. 
The other rat of group B’ (No. 53) was not killed. 

Post-mortem. Group A.—In rat No. 47 (killed June 14) the stomach 
is contracted and contains only mucus. Both the small and large intestines 
are congested. The pancreas is pink in colour. The contents of the large 
intestine are watery. In rat No. 48 nothing was noticeable. 

Group B.—In rats Nos. 49 and 50 the pancreas is fairly large and of 
a reddish-pink colour. 


Microscopic Examination of the Pancreas. Rat No. 47 
(thyroid feeding for twenty days, death from diarrhca),— 
The alveoli and their cells are generally small. The nuclei vary 
from 3u to 94%; most of them are comparatively large, and contain 
abundant coarse chromatin granules and enlarged nucleoli staining 
red by Mallory. Fully formed zymogen granules are only to be seen 
in a few of the alveoli (especially those near the islets). The granules 
in the rest of the alveoli are shown by Muir's method, but are not 
stained red by Mallory; they are scattered through the cytoplasm, 
and are not as usual confined to the inner zone. They perhaps repre- 
sent a pro-zymogen. There are no mitoses to be seen. Some of the 
cells contain vacuoles. 

Rat No. 48 (thyroid feeding for twenty-nine days).— 
Although a few of the alveoli are large, most are small; this applies 
also to their constituent cells. The nuclei vary in diameter from 
3°7u to 9u. The larger ones contain large nucleoli stained red by 
Mallory. There are no mitoses to be seen. Zymogen granules are 
relatively scanty, but form a distinct mass in the cells, many of 
which are vacuolated. 

Rats Nos. 49 and 50 (thyroid feeding for twenty-seven 
and twenty-eight days)—The description of the pancreas which 


Studies on the Endocrine Glands 293 


has been given for No. 48 applies generally for Nos. 49 and 50 (con- 
trol entire rats). These also show no mitoses. Zymogen granules 
are, however, abundant in all the alveoli, and vacuoles of variable 
size occur in many of the alveolar cells. 

Rats Nos. 51 and 52, 53 and 54, groups A’ and B’.—These are 
the animals which had received lean meat instead of thyroid, two 
having been castrated and the other two being entire. ‘The pancreas 
in all exhibits the normal microscopic appearance, the alveoli and 
the alveolar cells being of nearly equal size throughout, and the 
nuclei averaging about 5u. Zymogen granules are also fairly plenti- 
ful, and very few cells contain vacuoles. 


TaBLE XIV.—Errect or THyroip FEEDING ON CASTRATED Rats. 


A group B group A’ group oR group. 
(castrated). (entire). (castrated). (entire). 
Date, 
1916. 


No. No. No. No. No. No. No. No. 
47. 48, 49. 50. 51. 52: 53. 54. 


Body-weight in grm.| May 26} 130 | 150 | 130 | 130 | 175 | 185 | 280 | 210 


¥ Ss, UG) Se 135 115 120 190 145 290 215 
a , 22| .. | 147 | 110 | 115 | 190 | 150 | 310 | 220 
Weight of pancreas ee 06 o8 7 08 0-7 0°8 06 | Not | 09 
in grm. killed 
gare weight isis 1186 )L 21670 2163 \L 186 jl 2207 \1 2225) ... |L:283 
fe) pancreas to 


body - weight in 
grm. 


Summary.—Castration appears neither to have any distinct effect 
upon the structure of the pancreas nor to influence the effects of thyroid 
feeding. 

- 
DoEs THE ADMINISTRATION OF COMBINATIONS OF IODINE WITH PROTEIN 

AFFECT THE PANCREAS IN THE SAME WAY AS THYROID SUBSTANCE ? 


The administration by the mouth of a mixture (or combination) obtained 
from Parke, Davis & Co., and termed by them “thyro-protein,” and of 
a substance obtained from Martindale termed “iodo-protein,’ was next 
investigated. The 2-grain tablets of thyro-protein are stated to be 
composed as to 2 per cent. of their weight of a concentrated substance 
extracted from thyroid. lIodo-protein has the form of a brown powder 
containing about 10 per cent. of iodine, and having a slight odour 
of iodine. It is apparently an artificial compound. It is insoluble in 


294 Kojima 


water and dilute hydrochloric acid, but soluble in dilute alkalies. It is 
advocated as a substitute for alkaline iodides. 

One tablet of thyro-protein, or an equivalent weight of iodo-protein, per 
diem was ground up in a mortar with rusks, and a paste of the mixture 
was administered in the usual way to rats Nos. 56 and 57 (thyro-protein) 
and Nos. 58 and 59 (iodo-protein) during five days. During this period the 
animals exhibited no special symptoms; the appetite was normal, and there 
was no appreciable change in weight. 

On microscopic examination the usual effects of thyroid ad- 
ministration are seen in the pancreas of rats Nos. 56 and 57, 
which were fed on thyro-protein. A few cells show mitosis, but 
not as many as with the administration of whole thyroid. 
Zymogen is plentiful, and the granules are coarse. Some of the cells 
show vacuoles. No change is observable in the islets. The pancreas 
of Nos. 58 and 59, fed with iodo-protein, has practically a normal 
appearance. There are no mitoses, and the cells contain abundance 
of zymogen granules. 

Summary.—An artificial combination of iodine with protein does not 
cause the changes in the pancreas which are produced by thyroid, although 
these are produced by a natural combination of iodine with protein which 
is extracted from the thyroid. The effect is, however, less marked than 
when thyroid itself is used. 


WHAT IS THE EFFECT UPON THE PANCREAS OF IODIDES AND 
OTHER SALTS ? 


Sodium lodide. 


(a) Administration of Sodium Iodide to Normal Rats.—A small 
amount of sodium iodide dissolved in water was mixed with the food 
given to two full-grown male rats (Nos. 60 and 61), 0'1 grm. per rat per 
diem being administered in this way for five days. The appetite was 
somewhat affected, and diarrhcea was produced from the third day. The 
weights of the animals were respectively 145 grm. and 190 grm. at the 
commencement, and 135 grm. and 180 grm. at the end of the period, when 
they were killed. 

(b) Sodium Iodide Administration, with a Period of Inter- 
mission.—The same dose of sodium iodide was given each day for a 
week to two other full-grown male rats (Nos. 62 and 63: see Table XV.). 
Afterwards it was intermitted for seven days, and was then repeated 
during the succeeding week. From the fifth day of the first week the 
animals suffered from diarrhcea and appeared inactive. During the 
intermission they recovered. Throughout the second administration 
there were no symptoms to record. In all these experiments with sodium 
iodide there was an intensive iodine reaction in the urine, and this 


1 See W. H. Martindale and W. W. Westcott, The Extra Pharmacopeia, 1915, 
vol. i. p. 462. 


Studies on the Endocrine Glands 295 


continued until the animals were killed, in one case twenty-five days 
after the administration had ceased. (Rat No. 63 was killed seven days, 


rat No. 62 twenty-five days, after the second administration of iodide 
had ceased.) 


Post-mortem.—Rats Nos. 60 and 61 showed slight congestion of the 


intestines. In the others there was nothing noticeable. 


Pancreas.—On microscopic examination of the pancreas of all 
four animals considerable variation is seen in the size of the alveoli 
and of their cells (fig. 17). There are many more small alveoli than 


or 


* 7 f : } “« : F ; ; : 
Fig. 17.—Section of pancreas of rat (male) fed with an addition to its ordinary diet 
of 0°1 grm. of sodium iodide per diem during five days. Microphotograph ; 
magnified 400 diameters. Hematoxylin preparation. 
The appearances are similar to those of thyroid feeding. The alveolar cells 
vary remarkably in size, as do their nuclei. The latter contain abundance 
of coarse chromatin granules and are for the most part deeply stained with 


hematoxylin. Several of the cells exhibit mitosis. Some have abundance of 
zymogen, others only a little. 


in the normal condition, and there is a general increase in size of the 
cell-nuclei, although a certain number remain small. The nuclei are 
deeply stained by hematoxylin, and contain abundant coarse chromatin 
granules with large nucleoli; a few which are less deeply stained 
have only fine chromatin granules. Many of the nucleoli are stained 
red by Mallory. The zymogen granules are somewhat fewer than 
normal (fig. 18). A few cells contain vacuoles. In rats Nos. 60 
and 61 a certain number of the cells show mitotic figures and a few 
contain vacuoles, but in rats Nos. 62 and 63 neither mitosis nor 
vacuolation is to be found. 


(c) Administration of Sodium Iodide to Rats which had been 


previously thyroidectomised.—In this experiment four thyroid- 
ectomised rats were, after recovering from the operation, fed during five 


296 


Kojima 


Fic. 18.—Section of pancreas of rat (male) fed with an addition of 0°1 grm. of sodium 
iodide per diem to its ordinary diet during two periods of seven days, but with a 


week’s intermission. Microphotograph ; magnified 450 diameters. Mallory’s stain. 


Fic. 19.—Section of pancreas of rat (male), the thyroid of which had been removed, fed 


with an addition to its ordinary diet of 0°3 grm. of sodium iodide per diem during 
five days. Microphotograph ; magnified 90 diameters. Mallory’s stain. 

If this section is compared with the normal (fig. 4), it will be seen that there is little 
or no difference in the amount of zymogen in the alveoli. A section of an islet is seen 
in the preparation, and it is observable that as usual there is more zymogen in the 
alveoli adjacent to this than in the rest of the section. 


Studies on the Endocrine Glands 297 


days with the usual paste of ground rusks, to which 0°05 grm. of sodium 
iodide was added.! This experiment was later repeated on three other rats 
(Nos. 69, 70, 71), but with different doses. Rat No. 69, which received 
01 grm., suffered from diarrhcea from the second day onwards, with 
diminished appetite. Its weight at the end of the time was, however, 
hardly diminished. Rat No. 70, to which 0°2 grm. of sodium iodide was 
administered, showed a slight diminution of appetite and slight diarrhcea ; 
but again there was no change in the weight. Rat No. 71, which received 
0°3 grm. of sodium iodide, showed occasional diarrhcea and suffered con- 
siderable loss of weight, viz. from 170 grm. at the beginning to 150 grm. 
at the end of the experiment. In all cases there was strong positive evi- 
dence of iodine in the urine: it was first found in urine which had been 
collected nine hours after the commencement of the administration, but may 
well have been present sooner. 
Post-mortem nothing noticeable. 

Pancreas——The pancreas of rat No. 68 was not examined. In 
the others the appearances are nearly normal, zymogen granules 
being plentiful in all the alveoli (fig. 19). There is no evidence of 
mitosis, and very little sign of vacuolation. 

It follows from this that when sodium iodide is adminis- 
tered to rats deprived of the thyroid, the effects are not the 
same as when the thyroid is present. 


TaspLeE XV.—EFFECTS OF ADMINISTERING Sopium IopIpE TO Rats. 


Weight in 


Amount of . . nae 
ope food consumed CHE Bpecue Reaction of 
Date, 1916. |——_—___|_ per week in ear ce gravity of urine. 
No. | No. erm week in ¢.c. urine. 
62. 63. ; 
ee ee Ee 
July 19-25 310 | 325 210 200 1-060 Alkaline 
July 26-Aug.1} 315 | 325 230 195 1061 ee 
Aug. 2-8 290 | 305 160 210 1-060 4. 
0-1 grm. 
Nal per rat 
per diem 
9-15 | 295 | 306 170 200 1:058 z 
» 16-22 295 | 300 170 180 1-060 5 
O'l grm. 
Nal per rat 
per diem 
» 23-29 | 295 | 310 180 200 1-060 A 


oe se oe ee ee ee 

1 One of these animals (No. 68) died on the second day, probably from some accidental 
cause. It showed on post-mortem examination a considerable amount of general congestion 
of the intestines. 


298 Kojima 


Potassium lodide. 


An experiment similar to that with sodium iodide was made with 
potassium iodide, 0°1 grm. being administered to two rats (Nos. 64 and 65) 
for three and four days respectively, and to two others (Nos. 66 and 67) for 
five days. Beyond slight diarrhcea, which began on the fourth day, no 
special symptoms were observable, and there was no important difference in 


weight at the end of the time (Table XVI). 


Post-mortem.—The duodenum is congested in 64 and 65, but in the 


others there is nothing noticeable. 


TABLE XVI. 


KI for five | KI for four 


~ 


Killed on this date. 


Date, days. days. 
L1G.) | = Eee 
No. 66.|No. 67.| No. 65. 
Body-weight in grm. | July 20 17a) | W180 

s 33 ¢ 25 L7OE | USO? oe 

= Aug. 5 Be sa 180 

5 Pea) sti se os 

‘ Lo) “ps sh 180! 


KI for three 
days. 


No. 64. 


Fic. 20.—Section of pancreas of rat (male) fed with an addition to its ordinary food of 0°1 
grm. of potassium iodide per diem during five days. Microphotograph ; magnified 


400 diameters, Haematoxylin preparation. 


Most of the alveolar cells are small, but their nuclei are larger than normal; they 
stain deeply with hematoxylin. Many of them have central nucleoli, There is a 
great diminution in the amount of zymogen, which is absent from many of the cells. 


Studies on the Endocrine Glands 299 


Fig. 21.—Section from pancreas of rat (male) fed with an addition to the ordinary diet 
of 0°1 grm. of potassium iodide per diem during five days. This is from the same 
pancreas as that shown in fig. 20, but is stained with Mallory instead of hema- 
toxylin, and is magnified only 120 diameters. 

The section shows considerable diminution in the amount of zymogen as com- 
pared with the normal (fig. 4). 


Fic. 22.—Section from the same pancreas as that shown in fig. 21, and, like that, stained 
with Mallory, but magnified 450 diameters. 
The preparation shows the marked diminution in the amount of zymogen in the 
alveolar cells, in which, in consequence, the nuclei are more evident. The small, 
darkly stained bodies are red blood-corpuscles. 


300 Kojima 


Pancreas.—In the rats which were fed for five days (Nos. 66 
and 67) many of the cells are abnormally small, with comparatively 
large nuclei and a diminution of zymogen granules (figs. 20-22). The 
larger nuclei, many of which measure as much as from 9p to 11p, 
contain large nucleoli stained red by Mallory. The smaller cells 
contain numerous granules which are finer than the usual zymogen 
granules; they may represent a pro-zymogen. ‘These cells have the 
cytoplasm stained light red instead of blue by Mallory. There is no 
evidence of mitosis nor of vacuolation. 

In the rats which were fed for three and four days respectively 
(Nos. 64 and 65) the pancreas shows much the same microscopic 
appearance as the normal, with fewer large nuclei. There is no evi- 
dence of mitosis: some of the small cells are vacuolated. Zymogen 
granules are abundant in all the alveoli. 


Sodium Bromide, Sodium Chloride, and Sodium Fluoride. 


Similar experiments were made with these substances, an equal dose, 
viz. 0°l grm. per rat per diem, being administered during five days. Sodium 
bromide and sodium chloride produced no appreciable symptom. Sodium 
fluoride caused considerable effect upon the appetite; one of the two 
animals to which it was administered died on the fourth day. The other 
survived the experiment, and was killed along with the rest after five days. 

Post-mortem.—In the two rats fed with an addition of sodium fluoride 
there was general congestion of the intestines. In No. 77, the rat which 
survived the administration of sodium fluoride, the stomach was found to 
be contracted and to contain a thick mucous fluid and but little food. The 
contents of the small and large intestines were dark and watery; the 
intestines were congested but showed no ulceration. The liver and spleen 
were also congested. The pancreas appeared small, and pale in colour. The 
other animals show nothing noticeable. 


Pancreas.—The microscopic appearances are almost normal, but 
in Nos. 72, 73, 74, and 75, which were fed with addition of sodium 
bromide and sodium chloride, a few cells show mitoses, and one or 
two such were found in the section of the pancreas of No. 77 (sodium 
fluoride). Zymogen granules are plentiful in all the alveoli, and 
exhibit no difference from the normal. There is no vacuolation to be 
seen in the cells, nor anything to record about the islets. 


Sodium Carbonate, Sodium Phosphate, and 
Sodium Sulphate, 


Doses similar in amount to the salts already described were adminis- 
tered to a series of rats, and when these doses produced no obvious result, 
double doses were subsequently administered. 

Pancreas.—The effect of doubling the dose was to produce 


within the pancreas the appearance of a few mitoses, but no changes 
at all were apparent in the amount of zymogen, nor was there any 


_— 


Studies on the Endocrine Glands 301 


vacuolation in the alveolar cells, nor any appreciable change in the 
islets. 


Potassium Carbonate, Potassium Phosphate, and 
Potassium Sulphate. 


The above salts, even with double doses, fed to rats during five days 
along with their normal food, produced no special symptoms, and had no 
appreciable effect upon the weights of the animals. 


Pancreas.—On microscopic examination of the pancreas it is 
found that in nearly all cases the alveoli are small, and under a low 
power appear more compact than normal, The alveolar cells are also 
smaller than in the normal condition. In the rats fed with potas- 
sium carbonate (Nos. 84 and 85) the inner zone is faintly stained with 
hematoxylin and is not sharply distinguished from the outer zone. 
In all there are a few large nuclei, with abundance of coarse chromatin 
granules. The large nuclei are stained deeply in the hematoxylin 
preparations. In the potassium-carbonate fed rats some of the alve- 
olar cells have only scanty zymogen granules; most of the cells 
contain granules which are stained faintly red by Mallory; these are 
scattered in the protoplasm, and are not confined to the inner zone. 
In no case was there any evidence of mitosis, nor any vacuolation in 
the cytoplasm. 


Mercurie Chloride. 


This salt was administered (a) intravenously, and (b) subcutaneously. 
(a) 1 ec. of a 03 per cent. water-solution of mercuric chloride was 
injected into the caudal vein of a large male rat (No. 90, weight 190 grm.). 
Fourteen hours after the injection the animal appeared to be moribund; 
it was therefore killed and examined. 
Post-mortem there is general hyperzemia of the abdominal organs, 
especially of the intestines. 
Pancreas.—The pancreas shows, on microscopic examination, 
smaller alveoli than normal, giving a compact appearance under a 
low power, but there is otherwise not much difference from normal. 
There are abundance of zymogen granules and no evidence of mitosis. 
(b) Animal No. 91 (weight 150 grm.) received 1 c.c. of a 0-1 per cent. 
watery solution of mercuric chloride, administered subcutaneously. It was 
killed twenty-eight hours after the injection. 
Post-mortem the locality of the injection shows a certain amount of 
cedema. There is again general hyperemia of the abdominal organs, 
especially of the intestines. 


Pancreas.—The pancreas is reddish-pink in colour. The micro- 
scopic appearances are much the same as in 90, but some of the 
cells show mitoses. There is no indication of vacuolation. Zymogen 

1 It may be noted, with regard to the mitosis produced by corrosive sublimate in the 
pancreas, that this does not appear, as with thyroid feeding and with sodium iodide adminis- 


tration, to be confined to that organ ; for cells in other organs, such as the liver, ovary, and 
submaxillary gland, also exhibit mitoses after poisoning with this salt. 


302 Kojima 


granules are abundant in all the alveoli. In neither animal could 
any change be detected in the cells of the islets. 

Summary of the Effects of Salts.—Of the various salts examined, 
sodium iodide appears to be that which produces the greatest effect upon 
the pancreas, although a certain amount of effect is produced by other 
sodium salts. The most interesting change is the production of mitosis 
in the alveolar cells. It is noteworthy that this is not obtained when 
sodium iodide is administered to rats which have been previously thyroid- 
ectomised. It is therefore probable that the effect is produced by an 
excitation of the thyroid gland as the result of the administration. 


EFFECTS OF ADMINISTERING CERTAIN HORMONIC SUBSTANCES. 


Pilocarpine. 


For investigating the effects of pilocarpine two rats were taken, both 
non-pregnant females (Nos. 92 and 93), weight respectively 140 grm. and 
150 grm. Pilocarpine nitrate (0-006 grm.) in watery solution was injected 
hypodermically. The animals were killed after five hours. 


Pancreas.—The alveoli appear small, and under a low power the 
section looks compact. The alveolar cells are small. The cytoplasm 
is less dense than usual, and shows no sharp distinction between inner 
and outer zones. Some of the cells have small vacuoles. The nuclei, 
of normal size, contain fine chromatin granules. The nucleoli, which 
are not enlarged, are stained red by Mallory. There is no evidence 
of mitosis. Zymogen is remarkably diminished, and is scattered in 
the cytoplasm of the cells rather than being distinctly localised to 
the outer zone. It is much more abundant in the alveoli immediately 
surrounding the islets. The islet cells themselves are less dense in 
appearance, but their nuclei and granules seem normal. In spite of 
the intensive secretion which this dose of pilocarpine nitrate produces, 
and the consequent changes in the quantity of zymogen granules, 
there is no evidence of cell-division in the pancreas. 


Adrenalin. 

Two full-grown non-pregnant female rats (Nos. 94 and 95), weighing 
respectively 210 grm. and 200 grm., were used for testing the effect of this 
autacoid. To each animal was administered per diem 0'1 cc. of a Parke, 
Davis & Co.'s 1/1000 adrenalin solution along with their ordinary 
food. They exhibited no special symptoms during the administration. 
The appetite was not affected. The weight of the first rat was slightly 
increased and that of the second slightly diminished at the end of the 
experiment. 

Post-mortem there was nothing noteworthy. 

Pancreas.—On microscopic examination of the pancreas the 
alveoli and alveolar cells and their nuclei appear normal. The 


cytoplasm appears to have shrunk more than usual in the pro- 
cess of hardening, for around many of the nuclei a clear space is 


Studies on the Endocrine Glands 3038 


seen. There is no evidence of mitosis. Zymogen granules are far 
less numerous in all the alveoli than is the case in the normal 
pancreas. A few nuclei are larger than the rest, and some show a 
tendency to stain yellow instead of blue by Mallory. Some of the 
cells are vacuolated. The islets exhibit no appreciable change. 


Pituitary Body. 

The preparations of the pituitary body employed were (1) fresh ox- 
pituitaries, which were separated into anterior and posterior lobes, dried in 
an incubator at 37° C., and ground to a fine powder in a clean mortar. 
The powder was kept in an exsiccator. (2) Ox-pituitaries which had been 
preserved for a considerable time in chloroform. These were similarly 
separated into anterior and posterior parts, which were ground and pre- 
served apart from the others. 


(a) Feeding with Anterior Lobe. 

Four female rats (Nos. 96, 97, 98, 99), of weights varying from 160 grm. 
to 265 grm., were fed during a week with 0°3 grm. of dried anterior lobe. 
In one group of two the preparation from the fresh gland was used, in the 
other group the preparation from the chloroform-preserved gland. Alto- 
gether, during the period of feeding each animal of Nos. 96 and 97 
received 2°1 grm. during seven days, corresponding to about 10 grm. of 
fresh, undried anterior lobe. The animals showed no special symptoms 
when killed, after seven days, and their weights differed very little from 
those at the commencement. To the second group of two rats, Nos. 98 and 
99, a little more of the anterior lobe of pituitary body (that which had 
been preserved in chloroform) was given, and this feeding was continued 
for six days. The total amount which they received also corresponded to 
the anterior lobes of about ten fresh ox-pituitaries. In a third experiment 
as much as 1 grm. of the dried anterior lobe (chloroform-preserved) was 
given per rat per diem (rats Nos. 100 and 101, weights 180 grm. and 
190 grm. respectively). This treatment was continued for one week. No 
special symptoms were obvious, and at the end of the time the weights had 
not altered. 

Post-mortem there is nothing noticeable macroscopically. 

Pancreas.—The pancreas shows certain changes on microscopic 
examination. Thus the alveoli and alveolar cells are smaller than 
normal. The outer zone of each cell is relatively broader. There 
are quite a number of fairly large nuclei (754) containing large 
nucleoli staining red by Mallory. Zymogen granules are far scarcer 


than normal. There is no difference of appearance noticeable in 
the islets. 


(b) Feeding with Posterior Lobe. 


For this purpose dry posterior lobe of ox-pituitary was given to two 
groups of rats, that for the one group being derived from the fresh gland 
VOL. XI., NOS. 3 AND 4,—1917. 20 


304 Kojima 

and that for the other from the chloroform-preserved gland. In the 
former case 0°02 grm. and in the latter case 0°03 grm. of dry posterior 
lobe were given per rat per diem. The feeding was continued in each 
case for a week. No special symptoms were observed. The weights at 
the end of the experiment were increased in some of the rats and 
diminished in others, but not to any great extent. The amount of 
posterior lobe given per rat during the week corresponds to about ten 
posterior lobes of fresh ox-pituitaries. 

This experiment was repeated on two other groups of two rats, 0:03 
grm. of dry posterior lobe of ox-pituitary being administered to each—in 
the one case derived from the fresh glands, and in the other from the 
glands preserved in chloroform. The administration was continued for a 
week ; the weights at the end showed very little change. The object of 
repeating the experiment was to observe the effect, if any, upon the amount 
of urine, which I had omitted to notice in the preceding experiment. In 
these rats the amount of urine was greatly increased, the increase being 
much more marked in the animals to which the posterior lobe from fresh 
pituitary was administered than in those which received posterior lobe 
substance of glands preserved in chloroform: in these there was a little 
more increase in weight. The following table gives the quantities of urine 
passed in these animals, which were all males :— 


Taste XVII.—Rats FED witH PiturraRy Bopy. 


Fed with Fed with Fed with 
@outegl anterior lobe | posterior lobe duced ae 
: (preserved in | (preserved in ie 3 
terior lobe. 
Date, 1916. chloroform). | chloroform). 
| No. | No. | No. | No. | No. || No: Nomina 
TLS} L1G! |) 100. |) LOL |) Wd.) 12s) eS ee 
Body-weight in grm., Aug. 29} 175 | 180 | 180 | 190 | 180 | 170 | 210 | 220 
Body-weight in grm., Sept.5| 180 | 183 | 180 | 190 | 185 | 175 | 215 | 240 
Quantity of urine in a week 245 240 336 432 
im! ¢.c. 
Quantity of urine in a week 688°2 648°2 857-2 996°5 
per kilo of body-weight 
inse:¢. 
Specific gravity of urine 1-060 1:061 1-060 1:060 
Reaction of urine Alkaline Alkaline | Alkaline Alkaline 


It should be mentioned that the animals were freely supplied with water, 
besides that which they received along with the rusks, the water being 
offered from time to time but never left in the metabolism cage. In the 
urine of the rats which were fed with 0:03 grm. of dry posterior lobe 
(fresh), Nos. 118 and 114, there was a trace of glucose, as evidenced by 


Studies on the Endocrine Glands 305 


Fehling’s and Nylander’s tests, but no albumen. The rest showed neither 
albumen nor glucose. 


Post-mortem none of the organs of these animals (which were 
all fixed in 10 per cent. formol) showed a proper degree of firmness 
after fixation, apparently owing to a swollen and cedematous con- 
dition which can be noticed on microscopic examination to affect the 
cytoplasm of the cells. This applies not only to the pancreas but also 
to the pituitary body, the pineal gland, the thyroid and parathyroid. 
Otherwise there is nothing special to record, except that the zymogen 


Fic. 23.—Section of pancreas of rat (male) fed with an addition to its ordinary food of 
dry posterior lobe of ox-pituitary. Microphotograph ; magnified 500 diameters. 
Mallory’s stain. 

The zymogen in the cells is comparatively lightly stained by the acid fuchsin, 
and does not appear in the photograph in the form of black granules as in the 
normal gland. Notice the swollen condition of the alveolar cells due to a 
general vacuolation of their cytoplasm. 


granules in the alveolar cells of the pancreas stain less intensely than 
normally with Mallory (fig. 23). 


Summary of Effects of Pituitary Feeding on the Pancreas 
of the Rat.—Except that the zymogen granules appear to be more scanty 
than usual, feeding with anterior lobe of pituitary body seems to produce 
but little effect. Posterior lobe, on the other hand, causes swelling of the 
cytoplasm of the cells. The zymogen granules in the pancreas are not 
appreciably diminished in amount, but their character appears to be 
somewhat altered, for the material does not stain so intensely red with 
Mallory as is the case with the normal pancreas. The islets show no 
particular alteration as a result of either anterior or posterior lobe feeding. 
Feeding with anterior lobe has no effect upon the amount of urine, whereas 


306 Kojima 


feeding with posterior lobe causes a greatly increased quantity to be 
secreted. The results were much less marked with the posterior lobes of 
glands which had been preserved for a long time in chloroform. 


EXPERIMENTS ON THYROID FEEDING OF ANIMALS OTHER THAN Rats. 


Several previous workers have investigated the effects produced by 
thyroid feeding on animals other than rats. The following is a brief 
summary of their observations :— 


Berkeley (2), administering 5°15 grm. of fresh sheep’s thyroid 
to mice, found that they died in from three to ten days, with emacia- 
tion, rapid respiration, and tremors. The appetite decreased after 
two days’ feeding, and they lost body-weight. Carlson, Rooks, and 
M‘Kie (8) found that thyroid feeding produced in cats a gradual loss 
of weight, diminished appetite, and a tendency to diarrhcea and 
roughening of the fur. The loss in weight was rapidly regained on 
the cessation of thyroid feeding. Dogs either maintained their weight 
or increased it, but as young dogs appear to have been used, the gain 
in weight may have been due to normal growth. Most of the dogs 
remained healthy. Rabbits and guinea-pigs fed on thyroid lost 
weight rapidly. Diarrhcea was an almost constant symptom in the 
rabbit, less constant in the guinea-pig. These observers also fed 
chickens with 5 grm. to 65 grm. of thyroid for a long period (sixteen 
to ninety-two days). The general result was that these large doses 
caused loss of body-weight, and in all, or nearly all, toxic symptoms. 
They found carnivorous animals much more resistant than others, 
and correlate this with the higher percentage of protein in the 
food, suggesting that omnivorous and herbivorous animals which are 
accustomed to take less protein may be adversely affected by the 
addition of the amount of protein contained in the thyroid substance 
administered. The experiments of Caldwell (4) furnish evidence 
of the high resistance of dogs and cats to the specific autacoid of the 
thyroid. He suggests that the fatal results in rabbits might be due 
to susceptibility to foreign protein; but this seems improbable from 
the observations of French (5), that although ox-thyroid is poisonous 
to rabbits and guinea-pigs, a similar amount of material from the 
other organs of the ox produces no toxic symptoms. It is certain 
that the effects which have been produced in my experiments, 
especially the karyokinetic changes in the pancreas of the rat and 
the striking diminution of zymogen granules in all animals, cannot 
be due merely to excess of protein, for some of the controls received 
a large proportion of lean meat, and in these the pancreas remained 
perfectly normal. 


The following are the results of my own observations :— 


Effects of Thyroid Feeding on Mice. 
Two full-grown male mice (Nos. 1 and 2), weighing 25 grm. and 
22 grm. respectively, were fed with 0:2 grm. of dry ox-thyroid per mouse 
per diem added to the ordinary rusk paste. The feeding was continued 


Studies on the Endocrine Glands 307 


for five days (August 10-15). From the third day onwards the appetite of 
the animals decreased. At the end of the feeding they weighed 18 grm. 
and 20 grm. respectively. Two control mice on the same diet but without 
thyroid showed no appreciable alteration in weight. 

Post-mortem the intestines in both animals appear to be slightly con- 
gested. The stomach is full of food. 

Pancreas.—The alveolar cells are, if anything, larger than in the 
normal pancreas of the mouse. Many of the nuclei are enlarged, 
measuring from 7'54 to 10u. All aredeeply stained by hematoxylin, 
and contain coarse chromatin granules and large nucleoli staining red 
with Mallory. A certain number of the cells show mitoses. No 
vacuolation can be seen. There is a general diminution of zymogen 
granules, including the alveoli which are situated immediately round 
the islets. The islets are compact in structure, and are composed of 
cells which are nearly equal in size and appearance and contain 
uniformly sized nuclei. Their nucleoli are large, and stain red with 
Mallory. Their cytoplasm has fine granules which stain red by 
Mallory. The thyroid feeding does not appear to produce any 
change in the islets. 

Summary.—Thyroid feeding produces changes in the pancreas of the 
mouse similar to those which have been described in the rat. 


Effects of Thyroid Feeding and Thyroid Injection on 
Cats and Dogs. 

In the first instance, two full-grown male and two full-grown female 
cats were fed with an addition of sheep’s thyroid or dry ox-thyroid to 
their ordinary diet of bread and milk and fish or other meat food. The 
two female cats, Nos. 15 (pregnant) and 16 (non-pregnant), weighed respec- 
tively 2775 grm. and 2750 grm. They received 20 grm. of fresh thyroid 
per cat per diem.: the appetite of both animals decreased. On the third 
day of the feeding the pregnant cat gave birth to four kittens (apparently 
full time), which died shortly after birth. At the end of ten days’ feeding 
the cats weighed respectively 2690 grm. and 2700 grm. The two male 
cats were—No. 17, weight 3100 grm., and No. 18, weight 2750 grm. The 
former received 20 grm. of fresh sheep’s thyroid per diem for five days, 
the latter 17 grm. of dry ox-thyroid per diem for five days. In both the 
appetite was diminished after four days’ feeding. The weight of cat 
No. 17 was 3070 grm. when killed, that of No. 18, 2750 grm. (the same as 
at the commencement). A third male cat was injected subcutaneously each 
day during five days with a decoction of thyroid substance made from 
30 grm. of dry ox-thyroid. No special symptoms regarding appetite were 
observed in this animal. The weight at the commencement of the experi- 
ment was 3050 grm., at the end 2950 grm. The urine in this animal gave 
an intensive reaction when tested for iodine. 


Pancreas.—The alveolar cells are mostly of the same size as 
those of the normal cat’s pancreas. They are smaller than those 


308 Kojima 


of the rat; some of the cells show striation in the outer zone, which 
eannot as a rule be made out in the rat. The nuclei show evidence 


Fic, 24.— Section of pancreas of cat (male), meat-fed, normal. Microphotograph ; 
magnified 400 diameters. Mallory’s stain. 
The zymogen, which is stained deep red by Mallory, appears black in 
the photograph. 


Fig. 25.—Section of pancreas of cat (male), meat-fed, with an addition of 
20 grm. of fresh sheep-thyroid per diem during five days. Microphoto- 
graph ; magnified 400 diameters. Mallory’s stain. 

The tissue generally appears more compact than normal ; the most striking 
difference is the diminution in the amount of zymogen granules, which, even 
when present, are not so much stained by the acid fuchsin as in the normal 
condition. (Compare with fig. 24.) 


of enlargement, for whereas in the normal cat’s pancreas the nuclei 
measure from 5, to 6°2u,in the thyroid-fed animals the size varies 
from 54 to 7°5u, and some are as much as 8; but no evidence of 


ee 


Studies on the Endocrine Glands 309 


mitosis could be obtained, nor was there any vacuolation of the 
cells. The zymogen granules are, Rome’ er, diminished in amount as 
compared with the normal (figs. 24, 25). This is the case even ipo 
the alveoli immediately surrounding the islets, which in the cat, 

in the rat, show more abundant zymogen than the rest of ‘he 
pancreas. There is no appreciable difference to be seen in the islets 
as compared with those of the normal pancreas. 

Two experiments were made upon the dog. (1) A young male fox 
terrier (weight 4850 grm.) was fed with meat, porridge and milk, to 
which was added during ten days 13 grm. of fresh sheep’s thyroid per 
diem. This addition to the food produced no appreciable symptoms. 
At the end of the ten days the weight of the animal was 4950 grm. 


Fig, 26.—Section of pancreas of dog (male), normal. Microphotograph ; magnified 
400 diameters. Mallory’s stain. 
The zymogen granules are very abundant in the cells, and are stained dark 
red by the acid fuchsin, coming out black in the photograph. 


(2) The second dog taken, a non-pregnant female, was also a fox terrier 
(weight 6500 grm.). This animal, which was purely meat-fed (lights), 
received 39 grm. of dry ox-thyroid per diem during six days, in addition 
to its ordinary food. Again no appreciable symptoms were observable, 
and there was no perceptible change in weight. 


Pancreas.—Many of the alveolar cells appear small. Their 
nuclei are fairly large, and although the majority measure from 
5p to 6h, a considerable number are as large as 7'5u to 9u. There is 
no evidence of mitosis, nor is there any vacuolation of the cells. 
Zymogen granules are more scanty than in the normal dog’s pancreas 
(figs. 26 and 27 ). The islets show no change. 

In the dog also it is to be noted that zymogen granules are 
specially abundant in the alveoli immediately surrounding the islets, 
even in the normal animal. This difference is more apparent in 


310 Kojima 


the thyroid-fed dog, owing to the greater diminution of zymogen 
elsewhere. 

Summary.—Neither in the cat nor in the dog does thyroid feeding 
appear to cause cell-multiplication in the pancreas; at least no mitoses are 
observable after several days’ feeding, although in the rat they would be 
very numerous. The most prominent effect of thyroid feeding is a diminu- 


Fic, 27.—Section of pancreas of dog (male) fed with an addition to its ordinary 
diet of 18 grm. of fresh sheep-thyroid per diem during ten days. Micro- 
photograph ; magnified 400 diameters. Mallory’s stain. 

This section is to be compared with the preceding one. It shows a 
marked diminution in the amount of zymogen, the granules of which are 
not so darkly stained by the acid fuchsin as in the normal preparation. 
Some of the cells have very few granules. The nuclei are much more 
apparent than in the normal, owing to the part of the cell which con- 
tains them being free from zymogen. 


tion in the amount of zymogen in the alveolar cells. This applies to the 
cat which received the thyroid decoction by subcutaneous injection as well 
as to the animals which were fed by the mouth. 


Effect of Thyroid Feeding and Injection upon 
Rabbits and Guinea-pigs. 


All the animals employed were full-grown males. 

Rabbit.—Two rabbits received the pressed-out juice of 7 or 8 grm. 
of fresh sheep-thyroid per diem. For one of these (No. 1) the juice was 
smeared over the cabbage leaves which were given along with oats for 
fodder; in the other (No. 2) it was administered by means of a catheter 
passed down the cesophagus into the stomach. After two or three days 
the appetite in both was found to be considerably diminished, and in 
No. 1 there was severe diarrhcea on the fourth day of the thyroid feeding. 
Both animals became greatly emaciated, the weight considerably lessening. 
Both died on the night of the fourth day. A third rabbit, which was 


Studies on the Endocrine Glands 311 


injected subcutaneously with thyroid decoction, received the extract of 
3 grm. of dry ox-thyroid per diem. In this case again, on the fourth day, 
the animal was found to be inactive, with greatly decreased appetite. 
There was, however, no diarrhcea. Nevertheless, the animal died on the 
night of the fourth day. Post-mortem No. 1 showed a considerable 
amount of hyperemia of the intestines after death, and the contents of 
the large intestine were watery. The other two rabbits exhibited nothing 
special. No microscopic examination was made of any of these animals, 
since they had been dead several hours before the organs could be fixed. 

Guinea-pig.—Two full-grown males, weighing respectively 450 grm. 
and 560 grm., were fed with oats and cabbages during a week. After 
that period the pressed-out juice of 3 grm. of fresh sheep-thyroid was 
smeared on to the cabbage, which they ate as usual. (It was much more 
difficult to get rabbits to eat cabbage which had been smeared with 
thyroid juice.) One of the animals was found dead after four days, and 
the other after five days. In these cases also there was no microscopic 
examination of the pancreas. 


Effect of Feeding Birds with Thyroid. 


For this series of experiments specimens of the domestic fowl at various 
ages were employed. They were fed with a mixture of maize and broken- 
up rusks. 


It may be well first to give a short description of the normal 
pancreas of the bird. The pancreas of the domestic fowl shows 
certain peculiarities. As compared with the rat’s pancreas the 
alveoli are generally smaller, as are also the alveolar cells. 
The outer zone of each cell is narrow; it stains. deeply with 
hematoxylin. Most of the nuclei are small (3-7), but a certain 
number measure 54. The chromatin granules in the nuclei are 
fine. The nuclei are as usual placed near the base of the cells; they 
are for the most part only feebly stained by hematoxylin. Both 
nuclei and nucleoli are stained blue by Mallory. Although sections 
from a number of glands from normal animals were searched, only 
once could a mitotic figure be found. The cells do not show vacuo- 
lation. The zymogen granules vary considerably in amount. In 
some alveoli the cells are full of them, but there is no marked dis- 
tinction between inner and outer zones as in mammals. In other 
alveoli the zymogen granules are few, and are scattered within the 
cytoplasm. As has been noted in the rat, cat, and dog, zymogen 
granules are especially plentiful in the alveoli which immediately 
surround the islets: they decrease gradually in amount as the alveoli 
become more removed from those structures. Under a low power 
this produces in Mallory preparations the effect of red patches 
surrounding the islets. 

The islets are compact in appearance. Their cells contain very fine 
granules, red stained by Mallory, scattered uniformly in the cytoplasm. 
The nuclei measure 3°74, and contain fine chromatin granules. 


312 Kojima 


The above description applies to the pancreas.of the cock. In the 
pancreas of the hens examined both the alveolar cells and their nuclei were 
a little larger than those of the cock, and the zymogen granules much 
more plentiful. Otherwise there was no appreciable difference in the 
two sexes. 

In order to test the effects of thyroid feeding, two cockerels (Nos. 1 
and 2), weighing respectively 450 grm. and 460 grm., after having been fed 
for a week with maize and rusks alone, were fed, No. 1 with the addition 
of 8 grm. of fresh sheep-thyroid per diem for ten days, No. 2 with an equal 


Fig. 28.—Section of pancreas of cock, normal. Microphotograph ; magnified 
400 diameters. Mallory’s stain. 
The alveolar cells are many of them full of zymogen granules which are 
very darkly stained by the acid fuchsin. 


amount of lean mutton per diem for ten days. The appetite of cockerel 
No. 1 showed a decrease from the third day of the thyroid feeding on- 
wards; the animal became inactive, and showed slight diarrhcea (after the 
fourth day). Cockerel No. 2 exhibited no symptoms. At the end of the 
period of experiment their weights were respectively 440 grm. and 490 grm., 
the weight of the thyroid-fed one being somewhat decreased and the one 
without thyroid distinctly increased. 

Post-mortem the intestines of the thyroid-fed animal appear slightly con- 
gested when compared with those of the one which received no thyroid. 

A similar experiment was made upon two other cockerels weighing 
respectively 470 grm. and 395 grm., but with dried ox-thyroid; 45 grm. 
per cockerel per diem being administered during five days. The appetites 
of the animals showed a decrease on the third or fourth day, but there was 


Studies on the Endocrine Glands 313 


no diarrhcea. At the end of the time the weights were respectively 
450 grm. and 372 grm. There was nothing to be observed on post-mortem 
examination. 

For investigating the effect of injection of thyroid, a full-grown hen, 
weighing 860 grm., after having been fed with maize for a week, was in- 
jected subcutaneously each day with a decoction made from 5 grm. of 
dry ox-thyroid. On the third day the animal appeared inactive, and took 
very little food. The next morning it was found dead. Nothing could be 
noticed on post-mortem examination. 


Fic. 29.—Section of pancreas of cock fed with an addition to its ordinary 
food of 4°5 grm. of dry ox-thyroid per diem during five days. Micro- 
photograph ; magnified 500 diameters. Mallory’s stain. 

Notice the more compact appearance of the gland owing to the smaller 
size of the alveolar cells. There is marked diminution in the amount of 
zymogen, the granules of which when present are not stained nearly as 
darkly as in the normal pancreas. In many of the cells they appear 
merely as dark points scattered in the cytoplasm. 


Another bird, also a full-grown hen, weighing 950 grm., was then in- 
jected subcutaneously during five days with a decoction made from 7 grm. 
of dry ox-thyroid. The appetite was only slightly affected; there was no 
diarrhcea. At the end of the experiment the bird was killed; its weight 
was then the same as at the beginning. 

Pancreas.—Comparing the pancreas of the thyroid-fed and 
thyroid-injected animals with that of the normal animal (figs. 28 and 
29), it is to be noted that the size of the alveolar cells shows little or 
no difference; this is also true for their nuclei. Some of the nuclei 
have an irregular appearance suggesting mitosis, but we have been 
unable to assure ourselves that this is a karyokinetic appearance. No 
vacuoles are seen in the cells. There is a remarkable diminution in 


314 Kojima 


the amount of zymogen. No appreciable difference can be seen in 
the islets. 

Summary.—In the bird the mitoses in the pancreas, which in the 
rat are produced by thyroid feeding, have not been detected, but there is 
the same tendency to diminution in the amount of zymogen in the 
alveolar cells. 


GENERAL SUMMARY. 
Effects of Thyroid Feeding upon the General System. 


The effects upon the general system which in my experiments have 
resulted from thyroid administration are— 


(a) diminution of appetite and loss of weight ; 

(b) increased shedding of hairs ; 

(c) disturbances of the digestive system ; 

(d) diminution in both nitrogenous and gaseous metabolism. (This is 
dealt with later in a separate paper) ; 

(e) with prolonged feeding there may be a subsequent increase of 
weight, the appetite being regained. 


I have usually found that the appetite begins to diminish after three 
or four days’ administration, so that the animals take far less food (see 
Tables VI. and VII.) and the body-weight gradually decreases. Diarrhoea 
is sometimes present. Most of the animals become inactive. In rats the 
fur becomes rough, and the hairs fall out much more than normally. 
Throughout this period post-mortem examination shows congestion of 
the intestines, with soft or watery contents. In some cases the congestion 
is confined to the duodenum. 

When the thyroid feeding is continued for more than a week the 
symptoms gradually disappear. They are more marked with feeding 
than with subcutaneous injection of thyroid decoction. Possibly this is 
due to a smaller proportion of the effective autacoid being present in the 
decoction than in the whole thyroid. 

The above symptoms caused by thyroid administration have been noted 
by many previous observers. 


Presence of Iodine in the Urine. 


It is well known that iodine is almost always present in the thyroid, 
although there is marked variation in its amount in different individuals 
and in different species of animals. These variations probably depend on 
diet, but there are also seasonal and geographical variations—Martin (9), 
Guyer (10), Hutchison (11), Martindale (12), and others. I find that 
whereas the urine of the normal rat always gives a negative reaction when 
tested for iodine, the iodine test becomes positive after thyroid feeding and 
also after subcutaneous injection of thyroid decoction. In the latter case 
it appears about seven hours after the injection. If the thyroid feeding 


Studies on the Endocrine Glands 315 


has been maintained for several days and then left off, a positive result is 
still obtainable for at least as long as twenty-five days. A similar effect 
has been obtained after the administration of sodium iodide; the iodine 
showed itself nine hours after the dose had been given. 


Effects of Thyroid Administration on the 
Weight of the Pancreas. 


I have only investigated the weight of the pancreas in a few cases. In 
my earlier experiments I unfortunately omitted to determine this point. 
I find that after thyroid feeding to normal rats and also to castrated rats, 
either with an ordinary dose or with a somewhat increased dose, there is 
a marked increase in the size of the pancreas. 


Donaldson (6) does not give the weight of the pancreas in his 
work on the rat. Hoskins (7, a) determined the weights of the en- 
docrine glands of the new-born guinea-pig both normal and as 
affected by congenital hyperthyroidism caused by giving thyroid 
to the mother. He found the pancreas of the new-born animal 
under these conditions not demonstrably affected. In a second paper 
(7, b) he describes hypertrophy of the heart, liver, spleen, and supra- 
renals, and some loss of fat. Herring (8) finds that in thyroid- 
fed rats the weights of certain of the endocrine glands (the pancreas 
was not investigated) show an increase as compared with controls. 
He also noticed a great increase in the weight and volume of 
the heart. 


Histological Changes in the Pancreas produced 
by Thyroid Administration. 


These changes have been already briefly reported (1). Alterations in the 
pancreas are produced in all the animals investigated, but are by far most 
pronounced in the rat. The following is a brief summary of the effects 
produced :— 

The most striking result of thyroid feeding is multiplication of the 
alveolar cells, which soon become far more numerous and for the most part 
smaller than normal. During the first few days of thyroid administration 
there is clear evidence of karyokinesis; indeed, mitoses may be so abun- 
dant that one can make out in a field of the ordinary high power of 
the microscope (600 diameters) from three to six, and sometimes as many 
as from eight to twelve, cells in mitotic division; whereas, as is well 
known, the normal pancreas never or hardly ever shows mitoses. <Ac- 
companying these changes there is great diminution in the amount of 
zymogen within the cells. 

The cell-multiplication is still more marked when a second dose of 
thyroid is administered after a few days’ intermission, groups of small and 
evidently rapidly dividing cells being produced: in these groups the 
typical arrangement of alveoli is lost. 


316 Kojima 


If the thyroid feeding is continued over a long period (a month), most 
of the alveolar cells are again found to have become of the ordinary size— 
although a certain number of small cells may still be seen—and zymogen 
granules are again accumulated within the cells. 

The mitotic figures in the pancreas do not begin to appear until after 
three days’ feeding; after seven days’ feeding they become fewer in 
number and gradually disappear. They are still visible after three days’ 
intermission, following four days’ feeding, but after a week’s intermission 
are no longer visible. The mitotic figures are seen after fresh thyroid, 
dry thyroid, and boiled thyroid extract: also after feeding with the 
residue of the thyroid after a decoction has been made and filtered off. 
It follows therefore that the autacoid substance which excites changes 
in the pancreas resists the temperature of boiling water, and that it partly 
passes off into the water. It is, however, entirely insoluble in alcohol and 
ether, extracts made with these fluids producing no effect. 

Thyroid feeding also produces certain changes in the size and structure 
of the nuclei and in the staining properties of the nucleoli, which are 
described in the body of the paper. 

Another change which is produced is the formation of very distinct 
vacuoles within the cytoplasm; this vacuolation is not, however, general, 
but occurs in patches within the gland. Small vacuoles are occasionally 
to be seen in the normal rat’s pancreas, but those which are here referred 
to are large and conspicuous, and may occupy a great portion of the cell, 
displacing the nucleus towards the basement membrane. Here and there 
a communication can be made out between the vacuoles and the lumen of 
the alveolus.! 

No appreciable change can be made out in the cells of the islets of 
Langerhans nor in the centro-acinar cells. 


Effects of Castration. 


The effects of castration upon the pancreas have also been investigated ; 
the results have been negative. Previous castration does not affect the 
results of thyroid feeding. 


Effects of Iodides and other Salts upon the Pancreas. 


A further part of the investigation was directed to determining 
whether other combinations of iodine than that contained within the 
thyroid would produce similar effects upon the pancreas. With this object 
a combination of iodine and protein, sold under the name of iodo-protein, 
was compared with a commercial preparation obtained from the thyroid 
gland. The former was found to be ineffective, the latter effective. On 
testing the effect of administering certain salts, especially the salts of iodine, 


1 Vacuoles have previously been described in the alveolar cells of the dog’s pancreas as 
the result of vagus stimulation (Babkin, Rubaschkin, and Ssawitsch (13)). 


/ 


Studies on the Endocrine Glands 317 


the only one of these which seems to have the same effect as thyroid 
administration is sodium iodide, which tends to cause similar morpho- 
genetic changes in the pancreas, although of a less marked character. 
It appears to act by stimulating the thyroid of the animal, for when 
administered to rats which had previously been thyroidectomised, no effect 
was observed. 


Effects of Hormoniec Drugs and Autacoids upon the Pancreas. 


The effects of administering certain other substances which are known 
to possess the property of exciting glandular structures, viz. pilocarpine, 
adrenalin, and extracts of pituitary body, were also investigated. The 
results obtained with these are described. They are entirely different from 
those caused by thyroid feeding. 


Effects of Thyroid Feeding in Animals other than Rats. 


Experiments have been made to determine the results of thyroid feeding 
in a number of animals besides the rat, viz. mouse, cat, dog, rabbit, guinea- 
pig, and domestic fowl. Of these the mouse alone gives characteristic 
karyokinetic appearances in the pancreas like those described in the rat. 
In the cat, dog, rabbit, guinea-pig, and bird, mitoses have not been seen, 
but in all the disappearance of zymogen granules from the alveolar cells 
is marked. The general symptoms produced are similar to those seen 
in the rat. In almost all animals the appetite decreases during the 
administration. 


BIBLIOGRAPHY. 


(1) Kosima, Proc. Roy. Soc. Edin., 1916, xxxvi. 240; Proc. Physiol. Soc. in 
Journ. Physiol., 1916, vol. 1. p. xlv; and Communication to the British Association 
for the Advancement of Science, Newcastle-on-Tyne, 1916 (unpublished). 

(2) Berxetey, Johns Hopkins Hosp. Bull., 1897, viii. 137. 

(3) Caruson, Rooks, and M‘Krg, Amer. Journ. Physiol., 1912, xxx. 129. 

(4) CatpweLi, Amer. Journ. Physiol., ibid., p. 42. 

(5) Frencu, ibid., p. 30. 

(6) Donatpson, H. H., The Rat, 1915. 

(7) Hosxriys (a) Amer, Journ. Physiol., 1910, xxvi. 426-438; (b) Journ. of 
Exper. Zool., 1916, xxi. 295-346. 

(8) Herring, this Journal, 1917, xi. 47; also Communication to the British 
Association for the Advancement of Science, Newcastle-on-Tyne, 1916 (unpublished). 

(9) Martin, Pharm. Journ. and Pharmacist, 1912, lxxxix. 144. 

(10) Guysr, ibid., 1913, xci. 123. 

(11) Hurcutson, Journ. Physiol., 1898, xxiii. 179. 

(12) Martrnpate and Wesrcort, The Extra Pharmacopeeia, 1915, ii. 134. 

(13) Basxry, RupascHkry, and Ssawrrscg, Arch. f. mikrosk. Anat., 1909, Ixxiv. 68. 


318 Studies on the Endocrine Glands 


DESCRIPTION OF PLATES I. AND II. 


Puate I. 


A. Section of pancreas of normal male rat, Magnified 400 diameters. Stained 
by Muir’s method. 

B. Section of pancreas of male rat fed during seven days before being killed 
with addition of 1:0 grm. dried ox-thyroid to its ordinary diet. Magnified 400 
diameters. Stained by Muir’s method. 

A number of mitoses are visible in this section, whereas none are to be seen in 
the normal pancreas. The zymogen is greatly reduced in amount. 


Puate: If. 


Section of an outlying portion of pancreas of male rat, fed with addition to its 
ordinary food of 1:0 grm. dried ox-thyroid during one week, followed by a few days’ 
intermission, and then by another week of thyroid feeding. Magnified 600 diameters. 
Muir’s stain. Only one mitosis is seen in the field. The nuclei are very variable 
in size (in the normal pancreas they are very uniform); some are very large; for 
the most part they contain large red-tinged nucleoli. The cells also vary much in 
size: they have lost the regular arrangement into alveoh, and are separated by con- 
siderable spaces in which leucocytes and spindle-shaped cells (? connective-tissue 
cells) are visible. The zymogen granules are greatly reduced in amount. 


PLATE J, 


Quarterly Journal of Experimental Physiology.] [Vol. XI. 1917, 


Dr M. Kojima, “ Researches on the Endocrine Glands.” 


PLaTE II. 


Quarterly Journal of Experimental Physiology. ] [Vol. XI. 1917. 


Dr M. Kojima, “ Researches on the Endocrine Glands ” 


STUDIES ON THE ENDOCRINE GLANDS:—II.: THE RELA- 
TIONS OF THE PITUITARY BODY WITH THE THYROID 
AND PARATHYROID AND CERTAIN OTHER ENDOCRINE 
ORGANS IN THE RAT. By Masanaru Kovgima, Fleet Surgeon, 
Imperial Japanese Navy. (From the Physiology Department of 
Edinburgh University.) (With thirteen figures in the text and 


one coloured plate.) 


(Received for publication 1st December 1916.) 


CONTENTS. 
PAGE 
PREVIOUS OBSERVATIONS ’ / ; 319 
PRESENT INVESTIGATIONS. : 32] 
Microscopic appearances of pituitary of 1 rat ’ ’ 321 
Effects of thyroidectomy in rat . ; ; ; 324 
a parathyroidectomy in rat : 324 
a castration in the male rat. ; 326 
si feeding rats with thyroid and parathy roid. : 328 
a feeding rats with anterior lobe of ox- -pituitary ' : : 329 
af feeding rats with posterior lobe of ox-pituitary : 330 
és adrenalin administration in rat : ; : ; 330 
sodium and potassium iodides in rat . . , : 331 
Summary of observations on the rat. : 332 
Microscopic appearances of pituitary of dog. , é : 333 
Effects of partial thyroidectomy in dog . P ; 335 
_ parathy roidectomy in dog ; f ; ; 336 
Summary of observations on the dog. : , . : 337 
BIBLIOGRAPHY ; : , : 2 : : 337 


PREVIOUS OBSERVATIONS. 


THE present paper includes a study of the effects of thyroidectomy, para- 
thyroidectomy, and of castration on the pituitary body of the rat, and of 
partial thyroidectomy and parathyroidectomy on the pituitary of the dog. 
The effects of additions of thyroid, parathyroid, and pituitary body to the 
food were also investigated in the rat. 

Thyroidectomy.—Rogowitsch (1) was the first to examine the effect 
of thyroidectomy upon the pituitary. He has been followed by a number 
of other investigators, but their results are not in accordance. Most agree 
that the whole gland enlarges and increases in weight. Rogowitsch de- 
scribed the appearance of vesicles containing colloid in the pars anterior. 
His observations were confirmed for cases of myxcedema in the human 
subject by Sch6nemann (2), Boyce and Beadles (3), and others. Stieda 
(4) described the occurrence of enlargement of the so-called principal or 
chromaphobe cells (i.e. those staining but slightly with hematoxylin), and 
the appearance of vacuoles within them, whilst the chromaphil cells were 

VOL. XI., NOS. 3 AND 4.—1917. 21 


320 Kojima 


diminished in number. Herring (5) found that thyroidectomy causes en- 
largement of the cells of the pars intermedia, which stain more deeply than 
usual; in addition, the hyaline masses which occur in the pars nervosa are 
increased in amount. These changes are described in dogs, cats, and rabbits. 

It must, however, be borne in mind that most experiments on the sub- 
ject have been carried out without consideration of the fact that the para- 
thyroids were generally included in the operation. To investigate the 
influence of these, Cimoroni (6) performed total parathyroidectomy leaving 
the thyroid, and found that the pituitary body showed no changes; the 
dogs upon which he operated died, however, rather soon after the operation. 
Cimoroni also performed thyroidectomy alone upon a number of dogs, 
taking especial care to preserve the parathyroids. The animals survived 
for a considerable time after the operation, and he then found changes in 
the pituitary body similar to those which had already been described by 
other authors. He obtained similar results in rabbits. Harvier’s (7) 
experiments confirm those of Cimoroni. It would appear therefore that 
it is the thyroidectomy and not the parathyroidectomy which plays the 
more important part in effecting the changes which had been noticed in 
the pituitary after removal of both thyroid and parathyroid. 

Incidentally it may be remarked that if a part only of the thyroid be 
removed the rest enlarges and its epithelium undergoes proliferation (see 
succeeding paper). This compensatory growth of the remainder of the 
thyroid was noticed by Rogowitsch, and later by Gley and Nicolas (8), 
Horsley (9), Halsted (10), Edmunds (11), and Murray (12). Accord- 
ing to Halsted it is sufficient to prevent symptoms of athyroidism, even 
if only one-sixteenth of the original gland is left. 

Parathyroidectomy.—Regarding the parathyroids, Vassale and 
Generali (13), Moussu (14), Erdheim (15), Rouxeau (16), and others 
have found that although total parathyroidectomy in most animals pro- 
duces fatal symptoms of tetany, if two of the parathyroids are left there 
are usually no symptoms at all. If, however, three are removed, serious 
symptoms appear, although deferred. According to Alquier (17), 
Rouxeau, Harvier, Gley (18), Haberfeld and Schilder (19), after 
partial parathyroidectomy the remaining parathyroid undergoes hyper- 
trophy, the gland becoming heavier than normal. According to Pepere 
(20) and Vassale and Generali there is no cell proliferation, nor is there 
any obvious change in the structure of the cells, but a large amount of 
colloid accumulates in the lymph interstices of the gland and there is a 
tendency to the formation of vesicles in it. Haberfeld and Schilder 
state that no histological changes occur in the remaining parathyroid other 
than simple enlargement. 

Castration.—Fischera (21) was the first to notice in several species 
of animals (buffalo, bull, guinea-pig, rabbit, cock) that after castration 
the pituitary body enlarges to about twice its normal size and weight. He 
does not, however, state the body-weights of the animals operated upon, 


Studies on the Endocrine Glands 321 


so that we are unable to gather what the gain of weight of the pituitary 
body was in proportion to the body-weight, although it is evidently much 
larger relatively. He further found that on subcutaneous injection of 
testicular extract into the castrated animals the above changes in the 
pituitary did not appear, or, if they were present, disappeared. According 
to Cimoroni (22), the changes in the pituitary body which follow castra- 
tion are similar to those which follow thyroidectomy (dogs). Marassini 
and Luciani (23) found that in the sheep, domestic fowl, rabbit and guinea- 
pig the oxyphil (eosinophil) cells, which are normally abundant, exhibit 
little or no change in size and number, nor is any effect caused on the 
weight of the pituitary as the result of castration. They notice, however, 
an exception in the case of the cock, where, as the result of castration, there 
is a distinct enlargement of the pituitary, accompanied by the appearance 
of very voluminous cells the cytoplasm of which is granular and stains 
deeply with hematoxylin. They, however, regard these changes as 
essentially due to alterations in general metabolism and not a direct effect 
upon the gland. According to Kolde (24), in rabbits and guinea-pigs 
the pituitary body increases in weight after castration, and there is an 
accompanying increase in the oxyphil cells. Hatai (25) also found an 
increase in weight of the pituitary in male rats after castration. Living- 
stone (26), on the other hand, states that neither male nor female rats 
show any constant hypertrophy of the pituitary as the result of castration ; 
if anything, the pituitary is more often responsive in the female to the 
removal of the genital glands than in the male. Bied] (27), who gives the 
results of experiments carried out by Zacher] upon rats, states that the 
pituitary body undergoes enlargement after castration in both sexes. He 
finds the oxyphil cells relatively diminished in number; but big cells of a 
swollen character make their appearance. The nuclei of these cells are 
only faintly stained by hematoxylin: the cytoplasm is beset with fine 
granules and contains smal] vacuoles. Cells of this character are scattered 
either singly or in strands within the pars anterior. Bied] considers 
that these large cells are to be regarded as exhibiting end-stages of the 
process of secretion of the oxyphil cells. 


PRESENT INVESTIGATIONS. 
Microscopic Appearances of the Pituitary Body of the Rat. 


Under the microscope the pars anterior of the normal rat has a 
compact and very vascular appearance (fig. 1). When the vessels are 
filled with blood the capillaries are dilated so as to appear like sinuses. 
Oxyphil cells are present in great number in the pars anterior, mixed 
with other kinds of cells, and showing no distinct localisation. They are 
nearly uniform in size, but their shape varies considerably. Their cyto- 
plasm is compact and finely granular; their granules are stained red by 
eosin. Their nuclei, which are for the most part circular and uniform in 


322 Kojima 


size (5u), contain fine chromatin granules and are distinctly stained by 
hematoxylin. The cytoplasm of these cells is coloured red by Mallory 
(acid fuchsin), while their nuclei and the cytoplasm of the other cells of 
the gland are stained blue by this reagent. The other cells of the pars 
anterior are the basiphil cells and the principal or chromaphobe cells. 
The basiphils are fewer in number than the oxyphils, varying in size 
and in shape, some being much larger than the oxyphil cells. Their 
cytoplasm is less dense; it contains fine granules which stain faintly with 
basic dyes. The nuclei resemble those of the oxyphils. The so-called 
principal or chromaphobe cells occur in far greater number than the 


— = a - 
ger 
PB “ 
. i, 7 
+> 
= 


Fic. 1.—Section of pars anterior of pituitary body of male rat. Microphotograph ; 
magnified 500 diameters. Hematoxylin-eosin preparation. 
The cells which in the figure are a little darker than the rest are oxyphil cells. 
Several blood-vessels filled with blood-corpuscles are seen in the section. 


oxyphils and basiphils. The cytoplasm is only faintly stained by hema- 
toxylin. The nuclei vary in diameter from 5p to 5°5u. Vesicles of various 
size encircled by these cells are fairly frequent: some of the vesicles contain 
hyaline substance ; others appear empty. 

The pars intermedia is thickest opposite the middle of the cleft. Its 
cells are relatively small; their cytoplasm is finely granular and stains 
faintly with hematoxylin. The nuclei, which vary in size, are of much 
the same diameter as those of the principal cells of the pars anterior 
(5u to 5'5u). A few small vesicles encircled hy epithelium cells are 
occasionally to be seen embedded in the pars intermedia. é 

The pars nervosa has the appearance of a reticulum of fibres containing 
neuroglia-cells, with droplets of hyaline substance interspersed here and there. 


Studies on the Endocrine Glands 323 


Fi, 2.—Section of pars anterior of pituitary body of a male rat which had been subjected 
to thyroidectomy. Microphotograph ; magnified 500 diameters. Heematoxylin-eosin 
preparation. 


As compared with fig. 1, itis noticeable that many of the cells are enlarged. These 
enlarged cells are stained u faint pink by hematoxylin-eosin, and in the photograph 
appear lighter than the rest. Clear vesicles are also to be seen containing a thin hyaline 
material The darkly stained masses are blood-vessels full of blood-corpuscles. 


pt DY Ogg, Pr Ail 
Bern ¢ ie RA 


Fic. 3.—Section of pars anterior of pituitary body of rat (male) after parathyroidectomy. 
Microphotograph ; magnified 500 diameters. Hematoxylin-eosin preparation. 
Although most of the cells remain small, a certain number are greatly swollen, 
showing vacuolated cytoplasm and faintly stained nuclei. 


324 Kojima 

The intraglandular cleft is sometimes broad, in other cases quite 
narrow. Occasionally the pars anterior and the pars intermedia are in 
direct contact, so as to leave only a chain of spaces indicating the 
situation of the cleft. In other instances there is a wide space separating 
the pars anterior and pars intermedia. A small amount of hyaline 
substance may occasionally be seen within the cleft, but in many cases 


there is no such material visible. 


Effects of Thyroidectomy in Rat (fig. 2 and Plate III). 


After total thyroidectomy (the parathyroids were reimplanted in the 
wound, although I have not been able to obtain evidence that the graft took) 
striking changes become apparent in the structure of the pituitary body 
(fig. 2), provided the animals are kept sufficiently long after the operation.! 
Thus in an animal killed thirty-four days after thyroidectomy the pars 
anterior is less compact in structure than in the normal animal, and a large 
number of vesicles are visible over the whole section. These vesicles vary 
in size and shape. Many are full of hyaline substance which stains faintly 
red with eosin. Others are empty. There are also a considerable number 
of large swollen-looking cells the cytoplasm of which is open in appear- 
ance and is stained lightly by eosin: the outline of these cells is in most 
cases indistinct. Their cytoplasm contains numerous small vacuoles. In 
some of these cells the cytoplasm is coloured homogeneously red by eosin, 
the appearance being very like that of hyaline substance. All except the 
oxyphil cells are stained blue by Mallory (aniline blue). The hyaline 
substance just mentioned is also stained blue by Mallory, although, as 
already mentioned, faintly reddened by eosin in hematoxylin-eosin prepara- 
tions. The ordinary oxyphil and basiphil cells are remarkably few in 
number. The pars intermedia is relatively thickened. The above changes 
are already visible twenty-three days after thyroidectomy, and they are 
not removed after feeding the thyroidectomised animals for some days with 
1 grm. of dry ox-thyroid per rat per diem (fig. 6). 


Effects of Parathyroidectomy in Rat. 


Removal of parathyroid alone does not appear to be productive of such 
marked changes in the pituitary body as removal of thyroid. In the pituitary 
of a rat killed thirty-five days after parathyroidectomy, the pars anterior, 
which has a compact appearance under a low power, shows a certain number 
of small vesicles containing hyaline substance and a large number of oxyphil 
cells (fig. 3). These cells are for the most part slightly enlarged, and many 
of their nuclei are rather larger than in the normal gland (5p to 7-5y). 


There are a certain number of swollen cells similar to those described after 


thyroidectomy, but they are fewer. Their cytoplasm is spongy and finely 
vacuolated, and stains pinkish-red in hematoxylin-eosin preparations. 


! The rat is a particularly favourable animal for such experiments because it is not, as 
are most other animals, liable to tetany as an early result of removal of parathyroids. 


Studies on the Endocrine Glands 


: 
A ‘ 
” 


ew Als 
iP, 
“dF. : % : 
Ae Pe raft 


Fic. 4.—Section of pars anterior of pituitary body of a castrated male rat. Micro- 
photograph ; magnified 500 diameters. Hmatoxylin-eosin preparation. 
There are a number of characteristically enlarged cells with finely vacuolated 
cytoplasm and large, faintly stained nuclei, which contrast with the darkly 
stained nuclei of the smaller cells. 


b 


Fic. 5.—Section of pituitary of male rat fed with an addition to its ordinary food of 
1 grm. of dry ox-thyroid per diem during seven days. Microphotograph ; 
magnified 90 diameters. Hematoxylin-eosin preparation. 


a, pars anterior; 6, pars intermedia; c, pars nervosa; d, cleft partly obliterated, showing at 
one part an accumulation of hyaline substance. 


326 Kojima 


Some of these swollen cells appear to be undergoing degeneration, their 
nuclei being stained only faintly or hardly at all by hematoxylin. Basiphil 
cells are present in great number. Their cytoplasm is of an open char- 
acter and vacuolated. It is possible that the swollen cells above mentioned 
are formed by an alteration of some of these basiphil cells. 

A large number of the nuclei of the cells of the pars intermedia are 
enlarged (to about 7:5u). The pars nervosa shows comparatively few 
droplets of hyaline substance in its meshes. 


Effects of Castration in the Male Rat. 


From twenty to fifty-seven days after castration considerable changes 
are found in the pituitary body. The most marked effect is produced by the 


Fic. 6.—Section of pars anterior of pituitary body of thyroidectomised male rat fed with 
addition to its ordinary food of 1 grm. of dry ox-thyroid per diem during seven days, 
Microphotograph; magnified 500 diameters. Hzmatoxylin-eosin preparation. 

Many of the cells are greatly enlarged. There is a considerable accumulation of 
hyaline substance, apparently the result ofa degeneration of the cytoplasm, There are 
also a number of vesicles similar to those shown in fig. 2 as the result of thyroidectomy. 


appearance of a large number of swollen cells in the pars anterior (fig. 4). 
They are especially abundant near the periphery. Their description agrees 
generally with that of the swollen cells mentioned as the result of thyroid- 
ectomy and parathyroidectomy, so far at any rate as the cytoplasm is con- 
cerned. But many of their nuclei are much larger, and also have a swollen 
appearance: they measure from 57» to llw. Moreover, the chromatin 
granules of the nuclei, which are fine, are less distinctly stained by hema- 
toxylin than those of the other cells of the gland. These cells have an 


Studies on the Endocrine Glands 327 


appearance suggestive of degeneration, their outlines being in many cases 
indistinct. The oxyphil cells are large and plentiful. Their nuclei generally 
are enlarged, but not markedly. A few vesicles are to be seen containing 
hyaline substance. 

The pars intermedia is thickened as compared with the normal. Its 
cells are somewhat swollen. ‘The cytoplasm is vacuolated and the nuclei 


d 


a 


Fic. 7.—Section of pituitary body of parathyroidectomised male rat fed with addition to its 
ordinary food of 1 grm. of dry ox-thyroid per diem during seven days, Miecro- 
photograph ; magnified 90 diameters. Hematoxylin-eosin preparation. 

The section shows an accumulation of hyaline substance in the intraglandular cleft. 


a, pars anterior ; }, pars intermedia ; c, pars nervosa ; d, colloid in cleft. 


enlarged, measuring from 7'5~ to 1ly in diameter. The pars intermedia 
contains a few vesicles filled with hyaline substance. 

The masses of hyaline substance in the pars nervosa do not appear 
to be increased in amount. 

Table I. shows the weights of the pituitaries of four castrated male 
rats as compared with two controls. Three other controls are furnished 


in Table II. 


TaBLE I.—WEIGHT oF PituITaARyY Bopy AFTER CASTRATION. 


2 115. kre 
No. of rat : : : 110. jy 112. 113. (controls). 
Days after castration. 49 57 30 30 rer ae 
Body-weight in grm. 245 30) 315 310 265 250 
Weight of pituitary body ‘009 012 0097 ‘010 ‘008 ‘007 
in grm. 
Weight of pituitary body ‘037 040 ‘031 032 ‘030 "028 


per kilo of body-weight 
in grm. | | 


328 Kojima 


It will be seen that thirty days after castration there is no appreciable 
difference in weight as compared with the controls, but in the castrated 
animals which were kept forty-nine and fifty-seven days respectively there 


is a very appreciable increase. 


Effects of Thyroid and Parathyroid Feeding in Rat 
(figs 5) 16, 113), 

The addition of 1 grm. of dry ox-thyroid or 3-4 grm. of fresh sheep- 
thyroid to the ordinary diet for a week or more has the effect of producing 
diminution in number of the oxyphil cells of the pars anterior. Those that 
remain are for the most part swolien. The nuclei are not. much enlarged, 
but their chromatin granules are coarser and stain more deeply with hzma- 
toxylin. The cytoplasm of the basiphil cells is much more vacuolated than 
that of the oxyphils. Some of their nuclei hardly stain at all with hema- 
toxylin. The principal or chromaphobe cells are in far greater number than 
the rest. 

The intraglandular cleft shows a considerable amount of colloid sub- 
stance (fig. 5),and in one of the specimens large cysts, filled with a material 
of similar appearance, are present in the pars anterior. 

If the thyroid feeding is intermitted and then again resumed, or if 
continued for as long as a month, there is a gradual increase in the number 
of the oxyphil cells. There is also some appearance of swelling in the 
cells of the pars intermedia. When thyroid is fed to thyroidectomised 
and parathyroidectomised rats the changes caused by those operations 
are still evident, but there is a greater amount of hyaline substance in 
the gland (figs. 6 and 7). 

After feeding first with thyroid for a week and afterwards with 0-1 grm. 
of dry ox-parathyroid, the oxyphil cells again appear abundant. A few 
of these are swollen; in some of these swollen cells the nuclei are only 
very indistinctly stained with hematoxylin. Basiphil and chromaphobe 
cells are relatively fewer. 

The effect of thyroid feeding upon the pituitary of castrated male 
rats was also investigated. Some of the castrated animals were fed 
with addition of 3 grm. of fresh sheep’s thyroid to the food, and others 
with addition of an equal amount of lean flesh (mutton): these served 
as controls. The experiments lasted twenty and twenty-nine days 
respectively. 

In the animals fed with thyroid there are many large vesicles con- 
taining hyaline substance in the pars anterior, the appearance nearly 
resembling that observable after thyroidectomy (fig. 8). There are also 
a large number of swollen cells. Oxyphil cells are far fewer in number 
than basiphil. Many large drops of hyaline substance are to be seen in 
the pars posterior. 

Table Il. shows the weights of the pituitaries of four thyroid-fed 
castrated male rats as compared with three castrated controls. 


Studies on the Endocrine Glands 329 


Taste I].—Weicar or Pirurrary Bopy aFTER CASTRATION AND THYROID FEEDING. 


Group. . | Thyroid fed. Control. Thyroid fed. | Control. | 
No. of rat ; 17. 48. 49, 50. 61; |’ 62. 54. 
Days after castration | 20 | 29 a a. 29 29 
Body-weight in grm. | 110 | 145 110 115 190 150 220 
Weight of pituitary body | 0036 |~-0059 | -0034 | -0037 | -006 004 ‘007 

in grm. 
Weight of pituitary body | 033 | ‘041 033 032 ‘031 027 032 


per kilo of body-weizht 


The thyroid feeding does not appear to have had any influence on 
the weight of the organ, which is somewhat increased by castration alone 


(Table I.). 


Effects of Feeding Rats with Anterior Lobe of Ox-Pituitary. 


For this purpose dry anterior lobe of ox-pituitary was added to the diet 
of normal rats. The most striking effect which it seems to produce is the 


ae 


Fic, 8.—Part of section of pars anterior of pituitary body of castrated male rat fed during 
twenty-nine days with 3 grm. of fresh sheep-thyroid per diem added to its ordinary 
food.  Microphotograph ; magnified 500 diameters. Hematoxylin-eosin preparation. 

In this section several large cysts are seen containing a thin hyaline material. 
There are also some of the swollen cells described in the text. 


greatly increased number of oxyphil cells in the pars anterior—two-thirds or 
more of all the cells being of this nature—most of them being considerably 


330 Kojima 


enlarged. Their cytoplasm is compact: it is stained a deep red both by eosin 
and by Mallory (acid fuchsin). The nuclei of all kinds of cells present are 
somewhat enlarged, neasuring from 5p to 6; they contain abundance of 
chromatin. No distinctive changes are to be seen in the pars intermedia. 


Effects of Feeding Rats with Posterior Lobe of Ox-Pituitary. 


The addition of posterior lobe of ox-pituitary to the food produces striking 
changes in the pituitary body of the rat (fig. 9). The whole of the gland 
has a loose aspect. The pars anterior is characterised by the large number 
of oxyphil cells in it. The cytoplasm of these cells is open and finely vacuo- 


Fic. 9.—Section of pars anterior of pituitary body of a male rat which had been fed with 
addition of 2 centigr. of dry ox-pituitary (posterior lobe) per diem during seven days. 
Microphotograph ; magnified 500 diameters. Hematoxylin-eosin preparation. 

Notice the marked vacuolation of the cytoplasm of nearly all the cells, and the 
appearance of spaces between them, probably due to edema. There are also a 
number of clear vesicles in the section. The nuclei do not appear enlarged. 


lated. It is stained deep red both by eosin and by Mallory. The basiphil 
and chromaphobe cells are swollen and finely vacuolated. The nuclei of 
all the cells stain distinctly by hematoxylin and by Mallory (blue). Most 
of their chromatin granules are fine. The cells of the pars intermedia are 
swollen. The pars posterior has a looser appearance, as if there were an 
accumulation of fluid in the reticulum. Speaking generally, the cytoplasm of 
all kinds of cells in the gland is swollen, the nuclei being but little altered. 


Effects of Adrenalin Administration in Rat. 
After the addition of adrenalin to the food certain large cells become 
apparent in the pars anterior. These cells stain faintly red with eosin 
and deep blue with Mallory. Their cytoplasm is openly reticular, and 


Studies on the Endocrine Glands 331 


contains vacuoles of various size. They have large nuclei (94 to 10,), 
homogeneously but faintly stained by hematoxylin and coloured faintly 
blue by Mallory. These cells are for the most part much larger than the 
ordinary oxyphil cells, but are probably formed from them. They are 
scattered about, but are especially numerous in the middle of the pars 
anterior. Ordinary oxyphil cells are present in large number. Their 
cytoplasm is stained red both by eosin and by Mallory. Their nuclei are 
coloured distinctly by hematoxylin. Some are stained yellow by Mallory. 
Basiphil cells are also present in considerable number. Their cytoplasm 
is more open than that of the oxyphil cells. It is stained faintly by heema- 


Fie. 10.—Section of pars anterior of pituitary body of male rat fed with addition to its 
ordinary food of 0°1 grm. of potassium iodide per diem during five days. Micro- 
photograph ; magnified 500 diameters, Hematoxylin-eosin preparation. 

This preparation shows a large proportion of chromaphobe cells containing large 
nuclei with coarse chromatin granules. 


toxylin and by Mallory (blue). The pars intermedia shows a certain 
number of vesicles containing hyaline substance. There are, however, no 
distinctive changes in it nor in the pars nervosa. 


Effects of administering the Iodides of Sodium 
and Potassium in Rat. 


Marked changes occur in the pars anterior after administration of sodium 
iodide or potassium iodide (fig. 10). Under a low power the pars anterior 
is congested; the glandular substance has a compact appearance. There is 
a striking increase in the number of chromaphobe cells, with a similar 
reduction in the chromaphils (both oxyphils and basiphils). The cell- 


Son Kojima 


nuclei generally are larger than usual (54 to 7'5u). They contain abund- 
ance of coarse chromatin granules and stain more deeply than normally 
with hematoxylin. The oxyphil cells are far fewer in number than in the 
normal gland, and are smaller in size: their cytoplasm is compact. Scattered 
about in the pars anterior are numerous vesicles occupied by hyaline sub- 
stance, which is almost everywhere remarkably increased in quantity. The 
nuclei of the cells of the pars intermedia are also more deeply stained 
than usual by hematoxylin. The pars posterior appears more compact 
than in the normal gland: it shows abundance of hyaline droplets. . 

The above changes are rather better marked with potassium iodide than 
with sodium iodide. 


Summary of Observations on the Rat. 


1. The microscopic appearances of the several parts of the normal rat’s 
pituitary are described, with especial reference to the characters of the 
different kinds of cells they contain. 

2. The result of total thyroidectomy is to produce the appearance within 
the pars anterior of a number of large swollen cells with vacuolated proto- 
plasm, as well as a considerable increase of hyaline substance which is for 
the most part contained within vesicles in this part of the gland. The 
pars intermedia is thickened. In the rat I have not observed the increased 
discharge of hyaline substance through the pars nervosa described by 
Herring in the dog, cat, and rabbit. 

3. The result of parathyroidectomy is to produce the appearance of a 
few similar swollen cells; but the effect is far less than after thyroid- 
ectomy. The cells of the pars intermedia become enlarged. 

4. Castration in male rats is followed by the production of a large 
number of swollen cells in the pars anterior. The appearance of the cyto- 
plasm in these cells is very like that seen after thyroidectomy and para- 
thyroidectomy. But the outlines of the cells are indistinct, and their 
nuclei are much larger and contain fine chromatin granules; some of them 
show an appearance suggestive of degeneration. The cells of the pars 
intermedia are also enlarged and swollen. 

5. The addition of thyroid substance to the ordinary diet of normal rats 
causes at first diminution of the oxyphil cells in the pituitary body and 
accumulation of hyaline substance both in the intraglandular cleft and in 
the pars posterior. If the thyroid feeding is prolonged the number of 
oxyphil cells again increases: this is also seen if parathyroid is substituted 
in the diet for thyroid. In thyroid-fed, thyroidectomised, parathyroid- 
ectomised, and castrated rats the changes caused by thyroid feeding are 
still evident: indeed, there may be an even greater amount of hyaline 
substance produced within the gland. 

6. Feeding with anterior lobe of ox-pituitary causes an increase of 
oxyphil cells in the pars anterior. 

7. Feeding with posterior lobe of ox-pituitary produces a large increase 


Studies on the Endocrine Glands 333 


in the number of oxyphil cells. All the cells of the gland are swollen, and 
their cytoplasm has a loose open appearance ; the cell-nuclei are little altered. 
The pars posterior also has an open reticular appearance, as if there were 
an accumulation of fluid within it. 

8. Feeding with addition of adrenalin causes great enlargement of certain 
cells in the pars anterior: these seem to be the oxyphils. 

9. Feeding with sodium iodide or with potassium iodide causes an 
increase of chromaphobe cells with a concurrent reduction of chromaphil 
cells, both oxyphils and basiphils being involved. 


Microscopic Appearances of the Pituitary of the Dog. 


The pars anterior is compact in structure. Oxyphil cells are in large 
number, especially near the periphery: in fact, most of the cells of this part 


Fic. 11.—Section of pituitary body of normal dog (male). Microphotograph; magnified 75 
diameters, Hematoxylin-eosin preparation. 

The pars anterior contains numerous oxyphil cells which in the photograph appear dark. 
Numerous vesicles, either round or elongated, occur in the pars intermedia, which also con- 
tains an extension of the intraglandular cleft. The blood-vessels in the pars anterior are 
large and sinus-like, giving a peculiar mottled appearance to the section. 


" a, pars anterior; 6, pars intermedia, with cleft between them; c, pars nervosa. 


are oxyphil. They contain distinct granules, staining with eosin. Their 
nuclei are spheroidal, and measure about 54. The chromatin-granules 


334 Kojima 


are abundant, and are stained fairly deeply by hematoxylin. There is 
a smaller number of basiphil cells, the cytoplasm of which is less compact 
than that of the oxyphils. It is stained faintly by hematoxylin. Their 
nuclei are slightly larger, and contain fine chromatin granules. With 
regard to the principal or chromaphobe cells, their number and position 
vary considerably in different glands. There is some tendency to a vesicular 
arrangement. They have comparatively little cytoplasm, which is clear in 
appearance. Their nuclei are somewhat smaller than those of the oxyphils 


Fic. 12.—Section of part of pars anterior of the pituitary body of thyroidectomised 
dog (male). Microphotograph ; magnified 500 diameters. Hmatoxylin-eosin 
preparation. 

The preparation shows many enlarged cells the cytoplasm of which is finely 
vacuolated and which contain large nuclei, many of them irregular. There are 
also to be seen a number of clear vesicles and some clear cells with very faintly 
staining nuclei (? degenerating cells). 


(4. to 45). They have fine granules of chromatin. Between the cells is 
a small amount of reticular tissue with large, sinus-like capillaries. 

Pars intermedia.—The cells of the pars intermedia are in general 
smaller than those of the pars anterior, but vary in size as well as in 
shape. Where they line the cleft they are columnar. The cytoplasm 
is finely granular. The nuclei measure from 5p to 62; they contain 
abundance of fine chromatin granules. Many vesicles are observable in 
the pars intermedia, varying in size and shape. Each contains a hyaline 
mass. The cells which surround the vesicles tend to be columnar. 

Pars nervosa.—In the pars nervosa many small hyaline masses are 
seen scattered amongst the neuroghal fibres. 


b 


a 


Studies on the Endocrine Glands 335 


Effects of Partial Thyroidectomy in Dog. 


This operation was performed by Professor Schiifer upon a young male 
fox-terrier weighing 6400 grm. Care was taken to leave the parathyroids 
whilst removing as much of the thyroid tissue as possible, about two- 
thirds of the total thyroid tissue being removed; the portion of thyroid 
removed was, after fixation, submitted to section, and the series of sections 


Fic. 13.—Section of pituitary of a partially thyroidectomised dog (male) showing a large cyst 
containing colloid and several smaller ones in the substance of the pars anterior, and a 
considerable number of colloid-containing cysts in the pars intermedia. Microphotograph ; 
magnified 75 diameters. Hematoxylin-eosin preparation. 

The colloid within the cysts is stained faintly pink by hematoxylin-eosin. The sinusoid 
capillaries give a mottled appearance to the pars anterior. Notice an extension of the 
intraglandular cleft into the pars intermedia. 


a, pars anterior; }, pars intermedia; c, pars nervosa. 


investigated. They showed. no parathyroid. The wound had completely 
healed a week after the operation. The animal was fed throughout with 
dog-biscuits, except for a fortnight in December, when it was given horse- 
flesh, with the view of determining whether the parathyroids had been 
completely removed ; but in spite of the flesh diet no symptoms of tetany 
appeared. After the operation the animal’s weight increased, and continued 
to do so until it was killed—nine weeks after the operation,—when its 


weight was found to be 7450 grm. 
VOL. XI., NOS. 3 AND 4.—1917. 22 


336 Kojima 


Pituitary Body (figs. 12 and 13).—There is a large cystic cavity 
in the pars anterior communicating with the intraglandular cleft and 
containing a thin material which is stained faintly with eosin. The cavity 
is surrounded by cells belonging to the pars anterior and pars intermedia. 
Oxyphil cells are abundant in the pars anterior, especially in the most 
posterior portion and periphery. Most of them resemble the oxyphil cells 
of the normal gland, but some of their nuclei show shrinkage. Basiphil 
cells and chromaphobe cells are relatively few in number. 

A considerable number of cells are seen in the pars anterior, especially 
in the most anterior portion (but scattered also in other portions), which 
are many times larger than the ordinary cells of the gland. Even under 
a low power these cells are very apparent: they vary, however, in size and 
in shape. Their cytoplasm is open in appearance and finely granular. It 
stains faintly by hzmatoxylin, and contains numerous vacuoles. Their 
nuclei vary from 5» to 11» in diameter, and stain more faintly than those 
of the other cells. In many places these swollen cells surround vesicles, 
but there is no hyaline substance within these vesicles. 

Besides the large cyst in the pars anterior, other cysts are present, the 
hyaline contents of all being stained faintly by hematoxylin. The sub- 
stance has, however, a thinner appearance than the ordinary hyaline 
masses of the normal gland. 

The pars intermedia is thickened. It contains many vesicles of vari- 
able size, but most are larger than those normally found. They contain 
hyaline substance. The thickened pars intermedia with its vesicles extends 
here and there backwards into the pars nervosa. The hyaline droplets of 
the pars nervosa are not noticeably increased in amount. 


Effects of Partial Parathyroidectomy in Dog. 


For this experiment another young male fox-terrier (weighing 6050 grm.) 
was operated on, on the same day as the other dog. The two external 
parathyroids and the right internal parathyroid, with small portion of 
thyroid, were removed, the left internal only being left. Small pieces 
of both lobes of the thyroid were cut away, the largest portion for the 
sake of removing the right internal parathyroid (which was found on 
microscopical examination, after cutting this part into sections). The 
feeding and general circumstances of the experiment were similar to those 
of the other dog (partial thyroidectomy). On killing the animal at the 
end of the experiment (nine weeks) the weight was found increased to 
6850 grm. The animal suffered somewhat from diarrhoea, but showed no 
other adverse symptoms. 

Pituitary Body.—It is apparent even under a low power that all 
parts of the gland are much looser in texture than normal. The capillaries 
of the pars anterior are considerably dilated and full of blood. The oxyphil 
cells are remarkably abundant in all portions, without being distinctly 


ban a ee 


Studies on the Endocrine Glands 337 


localised to any one place. The pars intermedia shows numerous vesicles 
some of them containing hyaline masses. The pars nervosa is open in 
texture, and shows here and there the usual small droplets of hyaline 
substance: these do not appear increased in amount. 


Summary of Observations on the Dog. 


The effects of thyroidectomy—which was only partial, since it was 
desired to leave the parathyroids in situ—have been to cause changes in 
the pituitary consisting in the accumulation of colloid between the cells 
of the pars anterior, such as has been described by previous investigators, 
and an especial dey elopment of oxyphil cells, as well as the appearance 
of many swollen—possibly degenerating—cells, some of which are seen to 
surround clear vesicles. In the case described there was a cyst-like 
expansion of the intraglandular cleft in the pars anterior, but whether 
the result of the thyroidectomy or not, cannot be stated. The pars inter- 
media was thickened, and contained many vesicles, surrounded by cells, 
and occupied by masses of hyaline substance. These vesicles extend here 
and there into the pars nervosa. 

The chief effect of partial parathyroidectomy was to produce an 
cedematous condition of all parts of the pituitary. Oxyphil cells appeared 
more abundantly than usual in the pars anterior, but there were no such 
striking changes in the gland as are caused by thyroidectomy. 


BIBLIOGRAPHY. 


(1) Rocowirtscu, Centralbl. f. d. med. Wissensch., 1886, xxx. 530, and Ziegler’s 
Beitr., 1889, iv. 453. 
(2) Scuénemann, Virch. Arch., 1892, exxix. 310. 
(3) Boyce and Beapugs, Journ. Path. and Bact., 1892-3, i. 223 and 359. 
(4) Srizpa, Ziegler’s Beitr., 1890, vii. 
(5) Herring, Quart. Journ. Exper. Physiol., 1908, i. 281. 
(6) Crworont, Arch. ital. de biol., 1907, xlviii. 387. 
(7) Harvier, These de Paris, 1909. Cited by Guleke, Chirurgie der Neben- 
schilddriisen, 1913. 
(8) Gury and Niconas, Compt. rend. de la soc. de biol., 1895, 216. 
(9) Horsey, Brit. Med. Journ., 1896, 1623. 
(10) Hatsrep, Johns Hopkins Hosp. Rep., 1896. 
(11) Epwunps, Journ. Path. and Bact., 1898. 
(12) Murray, Lancet, 1899. 
(13) Vassate and General, Riform. med., 1897, ii. 631. : 
(14) Moussu, Compt. rend. de la soc. de biol., 1897, xlix. 44. 
(15) Erpuerm, Kongr. f. int. Med., 1906, Verhandl., 112. 
(16) Rouxeau, Compt. rend. de la soc. de biol., 1897, xlix. 17. 


338 Studies on the Endocrine Glands 


(17) Axgursr, Arch, de méd. expér. et d’anat. path., 1907, xix: Ob: 
18) Guey, Come rend. de la soc. de biol., LO9te 1S: 


0) PEPERE, Aves de med. ren et qaren oe 1908, 5.0.6 rile 
1) Fiscuera, Arch. ital, de biol., 1905, xli. 405. 

2) Crmoronl, op. cit. 

3) Marassini and Luctant, Arch. ital. de biol., 1911, lvi. 395. 
4) Koupe, Arch. f. Gynaek., 1912, Bd. 11. 1563, 

5) Harat, Journ. Exper. Zool., 1913, 111. 15. 

6) Livinestong, Amer. Journ. Physiol., 1916, xl. 153. 

( 


( 
( 
(: 
(: 
(: 
(: 
( 
(: 
(: 
(27) Brevi, Innere Sekretion, 1916, Teil 11. 108. 


DESCRIPTION OF PLATE III. 


Frontal section of part of pituitary body of thyroidectomised male rat, fixed 
with formol, cut from paraffin, and stained with hematoxylin and eosin. This 
animal before being killed had been fed for a week with addition of 1:0 grm. dried 
ox-thyroid to its ordinary food, 

a, pars anterior; 6, pars intermedia; c, pars nervosa. The pars intermedia is 
thickened, and encroaches considerably on the pars nervosa. It shows a number of 
cyst-like vesicles containing hyaline substance. 

Magnified 80 diameters. 


BR BD Rae BEB 


191 


(Vol. XI. 


Quarterly Journal of Experimental Physiology. | 


Dr M. Kojima, “ Researches on the Endocrine Glands.” 


STUDIES ON THE ENDOCRINE GLANDS —III.: THE EFFECTS 
ON THE THYROID AND PARATHYROID OF THE RAT 
OF ADMINISTERING THYROID EXTRACT AND CERTAIN 
OTHER AUTACOIDS AND SALTS. By Masanaru KoJima, 
Fleet Surgeon, Imperial Japanese Navy. (From the Physiology 
Department of Edinburgh University.) (With five figures in the 


text.) 
(Received for publication 1st December 1916. ) 
CONTENTS. 
PAGE 
INTRODUCTION. . , , : 339 
PRESENT INVESTIGATIONS. ‘ 340 
algae structure of thyroid and parathy roid of normal rat 340 
Effect of administering thyroid, parathyroid, and certain sodium and 
potassium salts : : 342 
Effect of administering posterior ‘lobe of pituitary body : 344 
Effect of administering adrenalin , ; ; 345 
SUMMARY ; ; : ; ; ‘ 345 
BIBLIOGRAPHY : . 345 
INTRODUCTION. 


BALLET and Enriquez (1) found no changes in the thyroid of dogs to 
which sheep-thyroid had been given in their diet. They also investigated 
the effects of injecting glycerine extracts of sheep-thyroid into dogs. Most 
of the animals died. In three they describe symptoms of experimental 
thyroidism. In two they notice enlargement of the thyroid. They also 
describe changes in structure, including the disappearance of alveoli in 
certain parts, with proliferation of their lining cells. Subsequently several 
authors, e.g. Lanz (2), Mediger (3), Georgiewsky (4), Cunningham (5), 
Gontscharnkow (6), and Ghedini (7) investigated the changes in the 
thyroid which follow administration of extracts to dogs, rabbits, and sheep. 
The results which they obtained are very various. Practically one may 
say that no constant changes were observed. Peiser (8) was more 
particularly interested in the study of the post-mortem changes in the 
nuclei in animals which had been either fed with thyroid or injected with 
thyroid extract. Most of the changes which he describes are accordingly 
post-mortem effects (autolysis). From his general results he concluded 
that when thyroid is administered by the mouth or subcutaneously to rats 
no special change occurs in the thyroid of the animal under observation. 
Chalmers Watson (9) found great variations in the structure of the 
thyroid both in the white rat and in the wild rat under varying conditions, 


340 Kojima 


but he arrived at the opinion that certain diets have a considerable effect 
in modifying the histological appearance of the gland, these modifications 
taking the form of changes in the amount of colloid or in its staining 
affinity with hematoxylin-eosin. He also described alterations in the size, 
shape, and number of the secreting cells, and in some instances noticed 
pronounced changes in the size of the whole gland. 

In the following account neither post-mortem autolysis nor the effects 
of alterations in diet upon the thyroid come into consideration, since the 
diet was constant and the animals were examined immediately after death, 
the thyroids being removed in the living condition and placed at once in 
10 per cent. formol. Nevertheless there is a certain variation in structure 
of the thyroid of the white rat (male), even when fed for a considerable 
time upon the same food. What this variation depends upon is not clear. 


PRESENT INVESTIGATIONS. 
Microscopic Examination of Thyroid and 
Parathyroid of Rat. 
Structure of the Normal Thyroid of the Rat.— Macroscopic- 
ally the gland varies considerably in size, shape, and colour. It is 


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Fie, 1.—Section showing adjacent portions of thyroid and parathyroid of normal 
male rat, rusk-fed. Microphotograph ; magnified 200 diameters, Hzmatoxylin- 
eosin preparation. 

The thyroid vesicles contain colloid which is only faintly stained. The 
lining epithelium cells are cubical. The parathyroid shows a comparatively 
compact structure. 


generally of a dirty pink. Its vesicles show considerable differences in 
size even in the same gland. Most are either spheroidal or oval; their 


eS eC 


oo od de al 


eee ee eee ee es) ee 


Studies on the Endocrine Glands 341 


colloid is stained faintly red by hematoxylin-eosin. The lining epithelium 
is composed of cubical or flattened cells, the cytoplasm of which is usually 
stained faintly by eosin. Some contain fine granules staining deeply red 
with eosin. The nuclei of the epithelium cells are round or oval (5+). 
They are usually in the middle of the cell, and are always distinctly stained by 
hematoxylin. They contain fine chromatin granules. Between the vesicles 
groups of epithelium cells are frequently to be seen; in some cases this 
may be due to the fact that the wall of a vesicle has been cut tangentially. 


Fie, 2.—Sections showing adjacent portions of thyroid and parathyroid of male rat 
fed with the addition of 1 grm. of dry ox-thyroid per diem to its ordinary food 
during two periods of seven days, with a week’s intermission. Microphotograph ; 
magnified 200 diameters. Hmatoxylin-eosin preparation. 

The thyroid vesicles are full of colloid, distinctly stained. Their lining 
epithelium cells are flattened ; some of the vesicles contain debris of epithelium. 
The texture of the parathyroid has a loose appearance owing to marked 
vacuolation of the cells. 


Peiser described the free end of the epithelium cells as melting gradually 
away into the colloid content of the vesicles; he states that the colloid 
material appears to pass between the cells to the blood-vessels. I have 
myself seen in some cases an appearance similar to that which Peiser has 
described, but in most instances the epithelium cells appear quite distinct from 
the colloid, and stain differently from it. There is, in point of fact, a well- 
marked boundary between the cytoplasm of the cell and the colloid sub- 
stance, and it is probable that the other appearance is due to the junction 
between the two being cut obliquely. 

With regard to the parathyroids. As was stated by Christiani (10) 
and by Erdheim (11), there is usually in the rat only one parathyroid 


342 Kojima 


on each side, embedded in the upper third of the thyroid substance near the 
surface. It can be seen even by the naked eye as a more opaque point in 
the substance of the thyroid. It would appear, however, that there are 
oceasionally aberrant parathyroids in these animals, and one is not perfectly 
certain that all parathyroid tissue has been removed even if the two obvious 
parathyroids are completely excised. 

Structure of the Normal Parathyroid of the Rat.—The para- 
thyroid consists of a large number of small cells which are usually com- 


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Fic. 3.—Section showing adjacent portions of thyroid and parathyroid of male rat 
fed with an addition of 0°1 grm. of sodium iodide per diem to its ordinary 
food during five days. Microphotograph; magnified 200 diameters. Hema- 
toxylin-eosin preparation. 

The thyroid vesicles are distended with colloid. Their lining epithelium 
cells are flattened, but rather less so than in the thyroid shown in the thyroid- 
fed animal (fig. 2). The parathyroid is less compact than normal (cf, fig. 1), 
but more so than in the thyroid-fed animal. 


pactly arranged. Each cell has a clear protoplasm which is comparatively 
small inamount. The cell-nuclei vary in shape, being round, oval, or oblong, 
but their size is fairly constant (about 5). They stain distinctly by 
hematoxylin, and contain fine chromatin granules. The cytoplasm is 
usually faintly stained by eosin. Occasionally there appears a vesicular- 
like arrangement of the cells, but this is rare. The parathyroid tissue is 
always quite sharply marked off from that of the thyroid, with connective 
tissue between. 


Effect of Administration of Thyroid and Parathyroid, 
and of certain Sodium and Potassium Salts. 


Changes in Thyroid.—After thyroid feeding most of the vesicles 
are large and distended with colloid. The epithelium cells lining them are 


Studies on the Endocrine Glands 343 


considerably flattened. The boundary between the colloid and protoplasm 
is less sharp. The cell-nuclei are deeply stained by hematoxylin. The 
distention of the vesicles with colloid gives a remarkably reduced appear- 
ance to the intervesicular connective tissue. The colloid is faintly stained 
by eosin. Occasionally debris of cells is to be observed mingled with the 
colloid. These changes become very pronounced if the thyroid feeding 
is continued for a long time. But there are individual differences, and 
Sag At Sra, oe eh MOF VO aT ae o> 
SERRE ATEN Re eS 


*yY PRIS 


Fic. 4.—Section showing adjacent portions of thyroid and parathyroid of male rat 
fed with an addition of 2 centigr. of dry ox-pituitary body (posterior lobe) to 
the ordinary food per diem for seven days. Microphotograph ; magnified 200 
diameters. Hematoxylin-eosin preparation, 

The epithelium cells lining the thyroid vesicles have a swollen appearance, 
and their nuclei are irregular, The colloid within the vesicles is thin; it is 
stained very slightly with hematoxylin. The parathyroid is not very different 
in appearance from the normal. 


in some subjects all the changes which have been described are far 
less distinct. 

After the administration of sodium and potassium iodide, changes in 
the appearance of the thyroid vesicles are observable which are for the 
most part similar to those recorded as the result of thyroid feeding. 

After feeding with parathyroid, there did not appear to be much 
difference between the thyroid of the animal and one fed with thyroid 
alone; but, since the animal fed with parathyroid had previously been 
subjected to thyroid feeding, it is possible that the effects of this had not 
passed off. 

Changes in Parathyroid.—lIt is not uncommon to find after thyroid 
feeding, as well as after administration of sodium and potassium iodides 


344 Kojima 


that the cells of the parathyroid no longer have a compact appearance, but 
are swollen, each containing a large vacuole. 

But occasionally this vacuolated appearance is observed in normal 
animals, although perhaps less marked than after thyroid feeding. It 
must be questioned if it is a specific effect on the parathyroid. 


Admistration of Posterior Lobe of Pituitary. 


Changes in Thyroid—Feeding with posterior lobe of ox-pituitary 
has a remarkable effect on the epithelium cells of the thyroid vesicles. 


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Fic. 5.—Section showing adjacent portions of thyroid and parathyroid of male rat to 
which 0°2c.c. of 1 per 1000 adrenalin solution was administered per diem during seven 
days. Microphotograph ; magnified 200 diameters. Hematoxylin-eosin preparation. 

The thyroid vesicles are filled with a thin colloid staining faintly by eosin. The 
epithelium cells are flattened, but much less so than with thyroid feeding. In some 
vesicles certain of the epithelium cells are swollen, and the swelling projects towards 
the colloid contents of the vesicle. The parathyroid has a loose open appearance : 
this seems to be due to swelling and vacuolation of its cells. 


Their cytoplasm is very clear, with a sharp outline stained faintly by eosin. 
They become more cubical, and acquire a swollen appearance: this gives 
the cells a barrel-like appearance. The nuclei are shrunken, and are 
uniformly stained by hematoxylin: they lie nearly in the middle of the 
cytoplasm. Many of the vesicles are empty of colloid. This, when present, 
is stained very faintly by eosin. In some of the vesicles debris of cells is 
seen mingled with a thin-looking colloid. 

Changes in Parathyroid—All the cells of the parathyroid are 
swollen, and the whole structure of the gland is loose, apparently as the 


ee 


Studies on the Endocrine Glands 345 


result of cedematous swelling. The nuclei, however, are shrunken, and 
stain darkly with hematoxylin. 


Administration of Adrenalin. 


Changes in Thyroid—Most of the cells of the vesicles are flattened. 
The vesicles are full of colloid, staining a delicate pink colour with hema- 
toxylin and eosin. Although in considerable amount there is less colloid 
than with thyroid feeding. It tends also to shrink more on fixation. 

Changes in Parathyroid.—Adrenalin seems to have much the same 
effect as posterior lobe of the pituitary upon the parathyroid, but the 
cells themselves are not swollen—indeed, the cytoplasm may be shrunken. 
All the cell-nuclei stain deeply with hematoxylin. 


SUMMARY. 


1. Administration of thyroid and of sodium and potassium iodides 
causes accumulation of colloid in the vesicles of the thyroid of rats. The 
epithelial cells lining the vesicles are flattened. The cells of the parathyroid 
are generally swollen. 

2. Administration of posterior lobe of pituitary gives the epithelial cells 
of the thyroid a swollen, barrel-like appearance ; their nuclei are shrunken. 
The structure of the parathyroid becomes loose. Its cells are also swollen 
and their nuclei shrunken. 

3. Administration of adrenalin causes somewhat similar changes in the 
thyroid and parathyroid to those produced by posterior lobe of pituitary, 
but they are less marked. 


BIBLIOGRAPHY. 


(1) Batuer and Enriquez, La médecine moderne, 1895, civ., and Sem. méd., 
1895, 330. 
(2) Lanz, Deutsch. med. Wochenschr., 1895, No. 37. 
(3) Mepicer, Dissert. Greifswald., 1895, cited by Peiser. 
(4) Grorciewsky, Zeitschr. f. klin. Med., 1897, xxxiii. 153. 
(5) Cunnineuam, Journ. Exper. Med., 1898, 11. 
(6) GontscHaRNkowW, Centralbl. f. allg. Path., etc., 1902, xii. 121. 
(7) Gueprnt, Centralbl. f. Bakt., 1903, xxxiv. 721. 
(8) Pxiser, Centralbl. f. allg. Path., etc., 1905, xvi. 513. 
(9) CHatmers Watson, Quart. Journ. Exper. Physiol., 1909, ii. 383, and 
1912, 229. 
(10) Curistiant, Compt. rend. de la soc. de biol., 1892, xliv. 798. 
(11) Erpuem, Ziegler’s Beitr., 1903, xxxili. 158, and Mitt. a. d. Grenzgeb. d. 
Med. u. Chir., 1906, xvi. 632. 


STUDIES ON THE ENDOCRINE GLANDS:—IV.: THE EFFECTS 
IN THE DOG UPON THE REMAINDER OF THE THY- 
ROID AND PARATHYROID OF PARTIAL REMOVAL OF 
THOSE ORGANS. By Masanaru Kogima, Fleet Surgeon, Imperial 
Japanese Navy. (From the Physiology Department of Edinburgh 
University.) (With two figures in the text.) 


(Received for publication 1st December 1916.) 


CONTENTS. 
PAGE 
STRUCTURE OF THE THYROID AND PARATHYROID OF THE Dog . : , 347 
OPERATIVE PROCEDURES AND THEIR EFFECTS : ; F : é 348 
SUMMARY 5 . . ; . : ; ; ; ‘ 349 


STRUCTURE OF THE THYROID AND PARATHYROID OF THE Doa. 


BEFORE describing the results of operations upon the thyroid and para- 
thyroid, the chief points regarding the microscopical structure of these 
glands in the normal animal may be enumerated. 

The Thyroid (fig. 1).—The vesicles vary in size and shape: they are 
lined by distinctly cubical epithelium. The cytoplasm of the cells is finely 
granular, and stains faintly with eosin. The cell-nuclei are round or slightly 
elongated, and of nearly equal size (5u). They contain fine chromatin 
granules, and most of them are deeply stained by hematoxylin. Between 
the vesicles is the connective tissue accompanied by blood-vessels. The 
vesicles are filled with a colloid which, in hematoxylin-eosin preparations, 
is stained of a faint red colour, with a nuance of purple. Between the 
vesicles are groups of epithelium cells; possibly some of these are due to 
walls of vesicles cut tangentially. 

The Parathyroids.—These are both internal and external. In 
number and position they show many departures from the type 
(M‘Callum and others). The internal are embedded in the thyroid 
substance, lying either in the upper or middle third of each lateral lobe. 
Both the internal and external parathyroids have a compact appearance, 
consisting of small epithelium-like cells. Their nuclei are about 5y in 
diameter, and contain fine granules of chromatin. The blood-vessels are 
convoluted. No vesicular arrangement of the cells is apparent. A thin 
capsule of connective tissue surrounds each parathyroid and separates the 
internal parathyroid from the thyroid substance. 


348 Kojima 


OPERATIVE PROCEDURES. 


The operations for removal of portions of thyroid and parathyroid 
have been described in a former paper, and the description need not be 
recapitulated. 


Microscopic Examination of the removed and remaining 
Thyroid and Parathyroids. 


Thyroid.—The removed thyroids are of normal appearance, and the 
description which has just been given will apply to them completely. 
Thyroid Remainders.—The size of these is obviously increased 


Fic. 1.—Section of normal thyroid of dog (male) removed by operation, fixed 
with 10 per cent. formol and embedded in celloidin. Microphotograph ; 
magnified 75 diameters. Hmatoxylin-eosin preparation. 

The vesicles are full of colloid, which is stained faintly by the hema- 
toxylin-eosin. The lining epithelium cells are cubical. 


if calculated from the size of the part removed. For although only 
about one-third of each lobe was left at the operation, what is now present 
would certainly be equivalent in size to at least one-half. 

Even the low power shows striking changes in the structure (fig. 2). 
The section appears like a number of tubules cut irregularly rather than 
a collection of vesicles. The size and shape of these tubules vary con- 
siderably. Generally speaking they are irregular, some of them branching. 
Many are empty, but some contain a small amount of a colloid which stains 
faintly red by eosin and has a much thinner appearance than normal. The 
columnar epithelium cells which line the vesicles are much higher than in 
the removed portions. Their cytoplasm is less compact, and contains small 
vacuoles. The granules within it are coarser than normal. Most of the 
nuclei are round. They vary in size from 3°7 to 6:24, and contain 


Studies on the Endocrine Glands 349 


abundant coarse chromatin granules. They stain for the most part 
very deeply with hematoxylin. Between the vesicles are seen consider- 
able groups of epithelium cells. In portions of the walls of the vesicles 
the epithelium cells have proliferated so as to cause a thickening and 
projection of the epithelium into the cavity. Here and there are 
mitotic figures. 

Parathyroids.—The removal of the parathyroids was confirmed 
by microscopical examination of the portions of the gland extirpated. 


Fic. 2.—Part of the gland which was left after removal of the larger portion. 

The animal was killed sixty-three days after removal. Microphotograph ; mag- 
nitied 80 diameters. Celloidin embedded, hematoxylin-eosin preparation. 

The vesicles are irregular in shape ; many of them are tubular. They vary much 
in size. The lining epithelium is thicker than normal There is a remarkable 
amount of intervesicular tissue which seems to be composed of the same cells as 
those which line the vesicles. The colloid within the vesicles has a thin appearance, 
and stains only faintly with hematoxylin. 


The remaining left internal parathyroid was examined after the animal 
had been killed, and showed no alteration from normal. 


SUMMARY. 


The remaining portion of thyroid after partial thyroidectomy under- 
goes proliferation of its epithelial cells and the vesicles show marked 
polymorphism. There is little or no change in the parathyroid. 

Partial parathyroidectomy causes no apparent changes in the remain- 
ing parathyroid tissue nor in the thyroid. 


STUDIES ON THE ENDOCRINE GLANDS:—V.: EFFECTS UPON 
METABOLISM OF CASTRATION, OF THYROIDECTOMY, 
OF PARATHYROIDECTOMY, AND OF THYROID AND 
PARATHYROID FEEDING. By Masauaru’ Koga, Fleet 
Surgeon, Imperial Japanese Navy. (From the Physiology 
Department of Edinburgh University.) (With thirteen tables 
and fourteen charts in an Appendix.) 


(Received for publication 1st December 1916). 


CONTENTS. 

PAGE 

LITERATURE . ; F : - , F ' ; , 351 
PRESENT OBSERVATIONS 2 ; : E ; ! ‘ : 353 
Nitrogen and calcium metabolism 3 ‘ : : 353 
Gaseous metabolism A 2 : . : t E : 358 
SUMMARY : : : ; : f ; ; ‘ : 359 
BIBLIOGRAPHY ‘ é ; - ? F ? : ; 359 
APPENDIX . : . i : ; oA) : : , 361 


Tue following observations are concerned with the effects of thyroidectomy 
and of parathyroidectomy, and of feeding with thyroid and parathyroid, 
upon the nitrogen, calcium, and gaseous metabolism; and with the effects 
of castration upon the gaseous metabolism. The animals employed in the 
course of these observations were all white rats, and are some of those 
which have been dealt with in the preceding papers. 


LITERATURE. 


Before describing the results of my own experiments, a brief account of 
recent literature may be given. 


Thyroidectomy.— Rovinsky (1) noticed, after the performance of 
thyroidectomy in male rabbits and dogs, that there is a diminution in both 
the gaseous exchanges and nitrogenous metabolism. He further found that 
on castration of thyroidectomised animals there is a still more marked de- 
crease of nitrogenous metabolism. Schneider (2) confirmed Rovinsky’s 
observations for female animals. He noticed the decrease of gaseous 
exchange and of nitrogen output in the urine to occur even before any 
cachexia had developed as the result of the thyroidectomy ; and during the 
period of cachexia thyreopriva both in castrated and non-castrated subjects 
the animals generally decrease in weight, with increased protein meta- 

VOL. XI., NOS. 3 AND 4,—1917. 


352 Kojima 


bolism. Mannsfeld and Miiller (3) found an increase of protein meta- 
bolism in thyroidectomised animals. They regard this as due to deficiency 
of oxygen and as related to the operation, since they found that a similar 
increase is produced by compression of the carotids. They were unable 
to observe any effect on their thyroidectomised animals as the result of 
feeding with thyroid tablets (Parke, Davis & Co.). 


Parathyroidectomy.—As a result of parathyroidectomy, Iselin (4) 
noticed an abnormality in the growth of the bones (rats). Erdheim (5), 
Hohlbaum (6), Toyofuku (7), and others have described insufficient 
calcification of the dentine, with hypoplasia of the enamel substance, in the 
teeth of rats. Morel (8) found that healing of bone fractures is delayed. 
M‘Callum and Voegtlin (9) noticed an increased output of calcium in 
the urine and feces in dogs during tetany, accompanied by a decrease of 
the calcium content of the brain and blood. Cooke (10), however, states 
that the calcium output in the urine does not increase in tetany, although 
he obtained augmentation of magnesium output. Noél Paton (10a) 
confirms Cooke with regard to the calcium content of the urine: he 
also finds that there is no appreciable change in the calcium content of 
the blood. Greenwald (11) states that the increase of nitrogen in dogs 
only occurs after the appearance of tetany following the parathyroidectomy. 
He found the ratio of urea N to total N to be decreased, whilst the pro- 
portion of N which was excreted in the urine in the form of NH, was 
very little if at all increased. He further obtained an increased elimination 
in the urine of nitrogenous constituents of an unknown nature (undeter- 
mined N). According to Cooke (12), both partial parathyroidectomy and 
partial thyroidectomy cause an increase of excretion of N and of NH, 
and the ratio of urea N to total N is diminished to a greater extent than 
is accounted for by an increase of NH, excretion. Cooke for the most 
part used fasting animals, while Greenwald’s animals were kept on a 
fixed diet. 


Thyroid an alee effects of thyroid feeding upon metabolism 
have been studied by a number of authors, but their results do not agree. 
In dogs most observers have found a decrease in body-weight (Scholtz (14), 
eo (15), Richter (16), Schéndorff (17), Georgiewsky (18), Gluzinski 
and Lemberger (19), Voit (20), Oswald (21), and others). Farrant (22) 
found loss of pene in cats and rabbits. Peiser (23) describes individual 
differences as occurring in rats. In one type there was an increase in 
weight; in a second the weight increased for some time, but after about 
four weeks of feeding it rapidly decreased until death; while in a third 
group the weight decreased gradually as the result of feeding, and there 
was no preliminary increase. When about one-third of the original weight 
was lost, death resulted. It occasionally happened that in the first two 
types there was slight decrease in weight for the first few days. Carlson, 
Rooks, and M‘Kie (24) obtained in rats a loss of weight as the result of 


Studies on the Endocrine Glands 353 


feeding (21 per cent. and 34 per cent. of the initial weight). Stoland (25) 
thought that the loss of weight which rats undergo within the first few 
days is partly due to accidental causes, such as handling, ete. Hewitt (26) 
also obtained loss of weight in rats as the result of thyroid feeding. 
Schafer (27) administered a small amount of thyroid (0°75-1°5 grm. of 
fresh sheep-thyroid) to growing rats, and found it to cause both an increased 
consumption of food and acceleration of growth. The experiments extended 
over three months. In a second paper Hewitt (26) also found that the 
administration of a small amount (0°25 grm.) of fresh thyroid (ox) caused 
an increase of weight in rats, accompanied by increase of appetite and food 
consumption. But when 0°5 grm. of fresh ox-thyroid was administered to 
adult white rats it produced loss of weight, the loss becoming greater as 
the amount of thyroid was increased, and being accompanied and _ possibly 
caused by diminution of appetite and lessened consumption of food. 


The above results appear to show that the effects of thyroid feeding 
upon metabolism depend partly upon the amount of thyroid administered 
and partly upon the duration of the feeding; perhaps also upon the 
amount of active substance contained in the thyroid employed. 


Castration.— Loewy and Richter (28) found in dogs that after 
castration there was a decrease in the amount of oxygen absorbed, this 
being more marked in females than in males. It was followed by laying 
on of fat. The result was confirmed by Pichtner (29), but according to 
Liithje (30) and Zuntz (31) there are occasionally found cases in which 
fat is not laid on. The experiments of Rovinsky and Schneider relat- 
ing to the effects of castration after thyroidectomy have already been 
mentioned. Shebuneff (32) obtained after castration in males (dogs and 
rabbits) an increase in body-weight and a diminution in the amount of 
nitrogen in the urine (decrease of protein oxidation). In most cases he 
noticed a decrease of CO, output and of O, intake. Cramer and M‘Call 
(33) studied the gaseous exchanges during five days of thyroid feeding. 
During the first two or three days the changes were slight, but after that. 
time there was a considerable increase in CO, excretion and oxygen 
absorption. When thyroid feeding was discontinued the gaseous meta- 
bolism returned gradually to the normal condition. The diet which they 
employed was one rich in carbohydrates (bread and milk). The amount 
of thyroid given was about 1 grm. of dried thyroid per diem. 

I may now proceed to give in detail the results obtained in the course 
of my own experiments upon rats. 


PRESENT OBSERVATIONS. 
Nitrogen and Calcium Metabolism. 


The animals employed in these experiments were adult white rats: three 
were females, all the rest males. They were kept in the special metabolism 


354 Kojima 


cages already mentioned (p. 257). Kjeldahl’s method was used for estimation 
of nitrogen; Cooke’s for calcium estimation in urine and feces. 

First Series.—In the first series of experiments eighteen male rats 
were employed. They were divided into three groups, A, B, and C, six to 
each group. From January 29 until March 10, 1916, they were fed with 
ground-up melox (Clarke, London), to which was added a certain amount 
of water, enough to convert it into a soft paste. Analyses of the melox 
used gave the following result :— 

In 100 Parts. 


Water , : : ; . 8086 
Ash . : 3 : : ac oO0 
Calcium 4 ; ; : SN Omiae 
Protein : : ; ; . 20°663 
abies : ; : : oo 
Starch : : } . 46°609 
Cane sugar . ; 7 858i 
Glucose : : : ‘ > sal 
Residue ! ; : t .  -Sials 


As will be seen from Tables I., II., and III., none of the animals showed 
any marked change in nitrogen and calcium metabolism during the first 
or normal week. On February 8 and 9 the animals of B group were 
subjected to parathyroidectomy and those of C group to thyroidectomy ; 
in the latter the parathyroids were separated from the excised thyroids 
and reimplanted in the depth of the wound, but I have no evidence that 
the graft took. The thyroid was completely removed, including the 
isthmus. After the animals had been killed the thyroids of Group B were 
cut completely in series, and it was thereby confirmed that no parathyroid 
had been left in the thyroid substance. The removed pieces were also cut 
in series, and proved to contain the whole of the parathyroid with a small 
piece of the adjacent thyroid substance. This of course does not exclude 
the possibility of accessory parathyroids having been left in the animals. 

For the first few days all the operated animals were inactive, and their 
appetite decreased. After a week the animals of B group (parathyroid- 
ectomy) were recovering, but the condition of those belonging to C group 
(thyroidectomy) was not so good, and the food was changed on March 11 
to a mixture of lean meat (60 per cent.) and rusks (40 per cent ), with 


sufficient water to make a soft paste. Analyses of this diet gave the 
following result :— 


In 100 Parts. 
(a) Fresh lean meat: 


Water ; : : i . 74186 
Ash , ; ; 3 ; a eOSS 
Calcium : 5 : : . 07028 
Protein : : : : . Q2aataes 
Fat. ; : j : v~ AC0Ss 


f Pick after filtering off the glucose, which was obtained in making the estimation of 
starch. 


Studies on the Endocrine Glands 355 


In 100 Parts. 
(b) Rusks: 


Water : ; : 6351 
Ash . : P } , 3°S60 
Calcium ; F . 0048 
Protein : ‘ 4 ; fy LL 
1 OF) a : ‘ , : ‘ 6°630 
Starch : : . 48921 
Cane sugar. ' ; : » 452) 
Glucose : é : : 2:982 
Residue ! : : ; : . 8580 


On March 25 the food was changed to rusks and water alone. From 
March 25 until March 31,1 grm. of dry ox-thyroid per rat per diem was 
added to this diet. From April 1 until April 7, 0°01 grm. of dry ox- 
parathyroid per rat per diem was added, the thyroid which they had been 
receiving being omitted. 

Some change had to be made in the number of rats in the three groups 
in the course of the experiment. Six male rats were included, as has 
already been stated, in A group and six in B group during the first three 
weeks, and the same number during the first two weeks inC group. From 
the fourth week until the eighth week there were only five rats in A group 
and five in B group, and after a certain time the number in C group was 
also different, since one of these animals died on the second day of the fifth 
week and another on the seventh day of the sixth week. Further, two rats 
of A group (the thyroid-fed unoperated control group) died during the tenth 
week—one on the first day and the other on the third day. 

The animals were always weighed at the same time of day, between 
3 and 4 p.m. The average body-weight is given in Tables I-III, (see 
- Appendix). After thyroid feeding there was a considerable difference in 
weight at the beginning as compared with the end of the feeding, but 
the full difference is not shown in the tables, since these give only the 
average weight during the week of feeding. 

After the eighth week one rat of each group was killed for histological 
examination. During the tenth week only one rat of A group, one of 
C group, and three of B group were available for the investigation of 
parathyroid feeding. 

Before operation the amount of food consumed by the three groups of 
animals showed no marked difference, and in A group (unoperated) it 
remained the same throughout. But after the operation differences in the 
amount consumed by B group and C group show themselves (Tables I.- 
III. and Chart 1). The amount of food consumed by B group rose 
gradually during the period extending from the fourth to the sixth week. 
Whether this was due to the recovery (rebound) from the diminished 
output immediately consequent on the operation, or to the actual removal 


1 Obtained after filtering off glucose converted from starch. 


356 Kojima 


of the parathyroid, is difficult to say. In C group the amount of food 
consumed remained much Jess than in A and B groups. The two animals 
of C group (thyroidectomy), which died during the fifth and sixth weeks 
respectively, did not appear to be ill, and took their food as well as the 
others until a few hours before death. 

During the seventh and eighth weeks, which was the period of. meat 
and rusk feeding, the amount of food consumed by B group (parathyroid- 
ectomy) diminished, while that of A and C groups remained much as it 
was before and after. When thyroid feeding was commenced the amount 
of food consumed diminished in all three groups: as a result the amount 
of nitrogen taken in was far less than before (Chart 2). During para- 
thyroid feeding the amount of food was increased in all, and the intake 
of nitrogen also was therefore increased. Since, however, the commence- 
ment of parathyroid feeding corresponded with the cessation of thyroid 
feeding, it may well be that the increased appetite was rather the result of 
the latter than of the former; in fact, it will be seen later in Series II. 
that parathyroid feeding does not itself appear to increase the appetite. 
During the period of feeding with combined meat and rusks (seventh and 
eighth weeks) there is, as might be expected, a large increase in the amount 
of nitrogen taken in in all animals; during it the animals of C group 
(thyroidectomy) showed no marked differences as regards the amount of 
nitrogen taken in as compared with Groups A and B. But during the 
period of thyroid feeding the amount of nitrogen taken in was far less in 
Groups B and C than in Group A. Two rats of Group A, however, died, 
apparently as the result of thyroid feeding; post-mortem there was con- 
siderable congestion of the intestines. The state of the heart was not 
noted, but Herring (34) has recently shown that sudden death is liable 
to occur in thyroid-fed rats, and that the heart in such animals becomes 
greatly hypertrophied. We may assume that these two (unoperated) rats 
were particularly susceptible to thyroid, while those which survived offered 
stronger resistance to it. In Groups A and B (normal and parathyroid- 
ectomised) no marked changes are noted in the amount of nitrogen in the 
urine up to the seventh week, while C group (thyroidectomised) shows a 
remarkable decrease. When meat was added to the diet a large amount 
of nitrogen appeared in the urine of all the groups, obviously due to the 
addition. During the thyroid feeding the amount of nitrogen in the urine 
showed diminution in all the groups, but this was especially marked in B 
and C. During parathyroid feeding, on the other hand, the amount of 
nitrogen increased in Groups B and C, while in A group there was, if 
anything, a little decrease (Chart 3). 

With regard to the amount of nitrogen retained in the body, in the 
animals of Group B there was less nitrogen retained during the second 
week, which might well have been due to the direct result of the operation 
(parathyroidectomy), but on recovery from the immediate results of the 
operation nitrogen retention was increased. In C group (thyroidectomy) 


Oe 


Studies on the Endocrine Glands 357 
the nitrogen retention does not show any marked diminution during the 
week following the operation, but it afterwards decreases considerably 
more than in the other groups. During the meat and rusk feeding a 
larger amount of nitrogen was taken in as food, but less was retained in 
the body. During the thyroid feeding the retention of nitrogen was far 
less in all the groups, but during the period which followed, in which the 
thyroid was omitted and parathyroid substituted, the amount of nitrogen 
again increased (Chart 4). 

Calcium.—During the first week of feeding with melox the amount of 
ealcium taken in was far greater than when meat and rusks or when rusks 
alone were used as food. This is accounted for by the fact that melox 
contains many small pieces of bone, and consequently far more calcium 
than the rusks or meat.! On substituting lean meat and rusks or rusks 
alone for the melox, the calcium contained in the food was therefore 
greatly diminished (Chart 5). 

With regard to the amount of calcium in the urine, it may be stated 
that there is always some daily variation in all animals. But although a 
diet of rusks or of meat and rusks contains so much smaller an amount of 
calcium than does melox, nevertheless there was no more calcium in the 
urine with the melox feeding than with the other diets. In B group 
(parathyroidectomy) the amount of calcium gradually increased, whereas 
in C group (thyroidectomy) it diminished (Chart 6). 

With regard to the retention of calcium in the body, it is noteworthy 
that no differential change is apparent as the result of thyroid feeding, a 
diminished retention occurring in all the groups. During the following 
week (parathyroid feeding) the amount again increased to its normal level 
(Chart 7), but again it is not easy to decide whether the increase of calcium 
retention was due to parathyroid feeding or to cessation of thyroid 
feeding. 

Second Series.—In this series of experiments three full-grown male 
and three full-grown female (non-pregnant) rats were employed. ‘The sexes 
were kept in separate metabolism cages. From May 5 to May 11 they 
were fed with rusk-paste alone. From May 12 to May 18, 0:1 grm. of dry 
ox-parathyroid per rat per diem was added to the food. From May 19 
to May 25 they were again fed with rusk-paste alone. From May 26 to 
January 1,3 grm. of fresh sheep-thyroid per rat per diem was added to 
the diet. 

During thyroid feeding the body-weight and appetite decreased, whilst 
during parathyroid feeding there was no marked change. During the 
parathyroid feeding the amount of food consumed by the females in- 
creased, while that of the males decreased. After the parathyroid feeding 
had ceased there was a remarkable increase in the amount consumed in 


1 Tt must, however, be stated that most of the bony fragments in the ground melox 
were rejected by the rats, so that the whole of the calcium in that diet was not ingested. 
It was partly on this account that it was discontinued and a meat and rusk diet substituted. 


358 Kojima 


both groups. During the period of thyroid feeding there was a decrease 
in the consumption of food in both groups. The amount of nitrogen taken 
in is strictly parallel to the amount of food consumed (Tables IV. and V., 
Charts 8 and 9). 

The amount of nitrogen in the urine is also parallel to the amount 
taken in: it is decreased during thyroid feeding (Chart 10). 

The retention of nitrogen in the body is also parallel with the amount 
of nitrogen taken in, and is decreased during the period of thyroid feeding 
(Chart 11). 

It would appear from the general results of this series of experiments 
that in unoperated animals the effect of parathyroid feeding upon nitrogen 
metabolism is negligible. 


Gaseous Metabolism. 


Third Series. After Thyroidectomy and Parathyroidectomy. 
—The animals belonging to A, B, and C groups of the first series were 
used in these experiments, the estimation of CO, being made twice on each 
group before the operation. For the estimation of CO, Haldane’s method 
was employed. The CO, output at the outset was very nearly the same 
in all the groups (Tables VI. to VIII., Chart 12). 

Several days after thyroidectomy had been performed on the animals 
belonging to Group C another estimation was made. These then showed, 
as compared with the animals of Groups A and B, marked diminution of 
CO, excretion; this was also apparent in the same (thyroidectomised) 
animals after several days’ feeding with meat and rusks. With that diet 
there is, however, also a diminution of the CO, excretion in the animals 
belonging to Groups A and B, but not so marked as with Group C. After 
six or seven days’ addition of thyroid to the diet there was in all groups a 
still further diminution of CO, excretion. On subsequently adding para- 
thyroid to the diet instead of thyroid, and continuing this for seven days, 
there is a small increase of CO, output in all the animals. It will, how- 
ever, appear from the next series of experiments that the augmentation 
of CO, output, which occurred after parathyroid feeding, was not due 
directly to that, but to the cessation of the thyroid feeding. 

Fourth Series. With Parathyroid and Thyroid Feeding.—-In 
another series of experiments (on the same groups of animals (three males 
and three females) as were employed in the N-metabolism experiment of 
the second series) the estimation of CO, output was made after five and 
ten days of thyroid feeding respectively. Two males were used as a control, 
these being fed on the same diet as the others, but without the addition 
of either thyroid or parathyroid (Tables IX. to XI.). (It may be noted 
that the amount of CO, excretion in female rats is always less than that 
of males under the same conditions of food, ete.) 

After five days’ thyroid feeding both the males and females showed a 
marked diminution of CO, output, while there is no difference after para- 


Studies on the Endocrine Glands 359 


thyroid feeding. After ten days’ thyroid feeding an increase of CO, output 
was apparent in the males, but a slight diminution in the females (Chart 13). 
These results appear to contradict those of Cramer and M‘Call, but there 
are not unlikely to be differences in susceptibility to thyroid feeding in 
different animals. 

Fifth Series. After Castration.—Twenty-two days and forty-eight 
days respectively after the operation the gaseous metabolism was investi- 
gated, first, in four, and, second, in two full-grown castrated male rats, and 
simultaneously in as many “entire” controls. All the castrated animals 
showed a marked diminution of CO, output, which continued up to forty- 
eight days after the operation (Tables XII. and XIII., Chart 14). In all the 
castrated animals the appetite and weight increased. 

It was noticed post-mortem that adipose tissue was remarkably developed 
in the subcutaneous tissue and at the back of the mesentery. 


SUMMARY. 


1. Thyroidectomy in rats produces a diminution both of nitrogen and 
of calcium output. 

2. After parathyroidectomy in rats there seems to be an increase of 
calcium in the urine, and less is retained in the body, but nitrogenous meta- 
bolism shows no definite change. 

8. Thyroid feeding produces a decrease in body-weight and a diminu- 
tion of nitrogen and gaseous metabolism in all the animals, whether normal, 
thyroidectomised, or parathyroidectomised. 

4, Castration is attended in male rats by diminution of CO, output. 


BIBLIOGRAPHY. 


(1) Rovinsky, cited in Zentralbl. d. exper. Med., 1914, 618. 
(2) Scunerper, Diss. St Petersburg. Cited in Biedl’s Innere Sekretion, 1916, 
Teil i. S. 196. 
(3) Mannsretp and Mtuisr, Pfliiger’s Arch., 1912, cxliii. 157. 
(4) Isexin, Deutsch. Zeitschr. f. Chir., 1908, xciii, 494. 
(5) Erpasrm, Frankf. Zeitschr. f. Path. 1911, vii. 175, 238. 
(6) Hontpavum, Ziegler’s Beitr., 1912, lil. 
(7) Tororuxu, Frankf. Zeitschr. f. Path., 1911, vii. 249. 
(8) Mors, Compt. rend. de la soc. de biol., 1910, lxviii. and Ixx. 780. 
(9) M‘Cartum and Vozertin, Johns Hopkins Hosp. Bull., 1908, xix. 91, and 
Journ, Exper. Med., 1909, xi. 118. 
(10) Cooxg, Journ. Exper. Med., 1910, xii. 45. 
(104) No&x Paton, Quart. Journ. Exper. Physiol., 1917, x. 
(11) GreenwaLp, Amer. Journ. Physiol., 1911, xxxvili. 103. 
(12) Cooxg, Journ. Exper. Med., 1911, xiii. 439. 


¢ 


360 Kojima 


(13) JuscurscHenKo, Biochem. Zeitschr., 1913, xlviii. 64. (Observations on 
thyroidectomy and parathyroidectomy and on thyroid feeding in dogs, dealing with 
the ratio of P to N as affected by those operations.) 

(14) Scuorrz, Zentralbl. f. inn. Med., 1895. 

(15) Roos, Zeitschr. f. physiol. Chem., 1896, xxii. 18, and 1898, xxviii. 40. 

(16) Ricursr, Zentralbl. f. inn. Med., 1896, 65. 

(17) Scuénvorrr, Pfliiger’s Arch., 1897, Ixvii. 385. 

(18) Grorerewsky, Zeitschr. f. klin. Med., 1897, xxxii. 153. 

(19) Guuziyski and LempereeEr, Zentralbl. f. inn. Med., 1897, 89. 

(20) Vort, Zeitschr. f. Biol., 1897, xxxv. 116. 

(21) Oswatp, Zeitschr. f. physiol. Chem., 1899, xxxvi. 39. 

(22) Farrant, Brit. Med. Journ., 1913, 11. 1363. 

(23) Prtser, Zeitschr. f. exper. Path. u. Therap., 1906, iii. 515. 

(24) Carson, Rooks, and M‘Kiz, Amer. Journ. Physiol., 1912, xxx. 129, 

(25) Srouanp, ibid., 37. 

(26) Hewirt, Quart. Journ. Exper. Physiol., 1914, viii. 113, 297. 

(27) ScuArsr, ibid., 1912, v. 203. 

(28) Loewy and Ricuter, Centralbl. f. Physiol., 1902, xvi. 449. 

(29) PAcurnsr, cited in Biedl’s Innere Sekretion, 1916, Teil ii, 273. 

(30) Lirxuse, Arch, f. exper. Path. u. Phar., 1902, xlviui. 184. 

(31) Zunrz, Zeitschr. f. Chir., 1908, xliv. 

(32) SHepunerr, Diss. Petrograd, 1914. Cited in Biedl’s Innere Sekretion, 
1916, Teil ii, 757. 

(33) Cramer and M‘Catt, Proc. Physiol. Soc., p. xxxvi (in Journ. Physiol., 
1916; x2) wand this. Journal; 1917 -.x1.759, 

(34) Herrine, Quart. Journ. Exper. Physiol., 1917, xi. p. 56. Also com- 
munication made to Brit. Assoc. for Adv. of Sci., Newcastle-on-Tyne, 1916 (un- 
published). 


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Seer eta lex Se ay) 2 Ol | ay 2 of : M4 rl oalliped o 5) o oH o a4 O oa : < 
JSSjPae eae) Se (SP Ses Ss] se] 8 4. 8| 4g Feels 2a as) sel sae (sg) sled | | ow 
2S hot ae (hg Mille 3) mB | aetinl leey lila. weet gy ape nila | | eS) s Ys |! 2s eel eee 2 3 
~ S bpi, BD] cm road = Peal Vt ta PL Saa7| BeEISOmS ewan ae ao mh mS | @ BO 

s 8 OS ee Orns Pe gaye afr ro a » ites OD EsglO 2 aa “4 5Sec0 an =] rid = 
= 5 StU! SS =] Sa a] Wee a fs IS > jae ESSE. a =| 1 eo Bes 30 2 iH &© rs) - 
ia" 16 ol 6 =i. | so oot! © agiaq ie TASASIS ES! SS! os Co |é8|s 2 
3 s6<|g-4 ee | gy Bs o| a Se! sg ce od SZR $o/8 ao a2 aan qo 5/15 = 
a if Asie ss] © Hes S bole SSsi§ BS3ie | da83l/4 [2 
< Be Mien VY) = e B = ct 9 ae 5 9 


‘(SLVY AasIKOLOAGIOUAH],) 9 daouy—']]] aAIavy, 


a 


Kojima 


364 


TaBLE 1[V.—MALE Rats. 


ae ‘((0°0) outa | 
jo yunoury 
"yystem-Apoq Jo 
opLy sad Apoq ut 
poureyar N [eIOL 


1:986 | 105 


96 


‘saomy pure oun 
EINE LIND 


UN [ROL 


2°786| 6°201 


‘qB1aM-Apog 
jo opty aad outa 
NEI o 


od 
‘ 


5944 | 3°079] 6°277| 2°315 | 109 


5148 | 2355] 4°826| 0°973 


‘ouTan 


UL NC PIO, 


"4yStam-Apoq Jo 
o[Ly ted poumnsuoo 
POOP ME NGO 


| 


“‘pouINsuod Poo} 
TUN [3908 


243) 13°796) 3°415) 6°42 


‘ 


7°104| 13-480) 3°309} 6-279 | 2°686| 5°995| 2°105 | 111 


"yy Stom-Apoq Jo 
ory aed pournsu09 
poo} Jo yunoury 


483°8 


*petansuoo 
poo} jo yunoury 


260 | 495°2 


255 


‘syed JO aquinyy 
‘dnoas 
JO JYSIAM [B10], 


3 


3 


270 | 501°8 | 7°522) 13°981)| 3:198 


3 


190 | 395°8 | 5-293) 11-271) 2°471 


3 


25 


Sutpus Yao Ah 


1916. 


18) 527 


” 


25) 538 


” 


"HO2,A\ 


I. |May11 


IN 


Inne 


IV. |June 1} 480 


TABLE V.—FEMALE Rats. 


‘(o0) oun 

jo qunoury | 
‘qysiem-Apoq Jo 
opey aed Apoq ut 
Peareyet NN TeoL 


2°309 | 115 


89 


‘soo@y] PUP OULIN 
NE east 


“‘So0a} | 


UT NI [VIOL 


2:044) 4°464 


2°278) 4-600} 0-862 


"qystom-Apoq 
yo opty aed outa 
ul N [®IO], 


4:829 


Ut N [°9O, 


‘qyustam-Apoq Jo 
opLy ted pourmsuo0o 
pooj Ut N [@I0,7, 


14°634| 3:092| 6:017 


“pettinstoo 
poo} UL N [P90.L 


5°014| 10°424) 2°322 


‘qystam-Apoq Jo 
ory aed petunst09 
poof jo JuNoW Wy 


74:2 


*pouinstoo 
poof jo qunowmy 


230 | 455-4 | 6°407| 12°489) 3:024) 5-989 | 2°3'70/ 5:394) 2-007 | 110 


240 | 474°3 | 6°686| 13°213] 3°112| 6°152 | 2-494) 5-606} 2-135 | 107 


180 | 3 


‘syvx JO aQuUINN 


3 


3 


‘duoas 
JO JYSIOM [B10], 


505 


SUTPUA YI AA 


1916. 
Mayll 


v 


June 1} 481 


"00M 


iff 


INE 


IV. 


g the fourth week 3 erm. of fresh sheep- 


During the second week 0-1 grm. of dry ox-parathyroid 


per rat per diem was added to the food. Durin 
per rat per diem was added to the food. 


All were fed with rusk-paste. 


YY 


thyroid 


Studies on the Endocrine Glands 365 


Taste VI.—CO, Excretion. 


Group A (Controls, Male). 


- | =. | ry) 
a3 ; | Sos | a6 tog | & 
els) 2 | 25? | o82 | S35] : | | 
3 ~ = | Fen | Ofc] ac =y Date. Food. Remarks. 
= , x“ +S » = 7 = 3) | 
Bele | 553) P85 | 28] = | 
m in| = es 290] ne = 
4"s l|aa| @ 
es = 2 aes i ee eet ieee 
1 | 260 | 4-099 °F. | 1916. | | 
2 | 180 4519 | 
I : a ee - 4455 15 53 Jan. 29 Melox Room is heated 
5 | 205 4162 
6 | 115 4324 
! 
1 | 265 4-075 
2/180} 4611 | 
Il. ? as aes , 4462 15 52 Feb. 5 Melox Room is heated 
5 | 205 4°128 
6 | 120| 4-296 | 


1 | 265 4059 
2 | 180 4644 | 
III. 3 | 185 5°329 |; 4502 15 48 Mar. 8 Melox 
4 | 235 4°412 
5 | 205 | 4068 


/ From Mar. 11-24:| Tenth day of 


1 | 260 | 3°303 
2} 180] 4531 | 


: Y rusks 40 per cent. rusks and lean 
IV. A oe ate | 4205 15 48 | Mar. 20 | and lean meat 60 meat feeding 
5 | 205 |. 4-972 | Pee 


21135 | 3°407 | | | From Mar. 25-31: | Seventh day of 
{2 |285| $24 |Leew | a0 | oe farm || serie] it te 
| 5 | 160} 3-712 per rat per diem 


Ns 


From Apr. 1-7:| Seventh day of 
; | rusks + 0°1 grm. parathyroid 
VI. | 3 | 165 | 4-242 4242 15 50 4 Apr 7 + of dry ox-para-| feeding 
thyroid per rat 
per diem 


366 Kojima 
TasLe VII.—CO, Excretion. 
Group B (Parathyroidectomised Rats, Male). 
Device anes o | 
4 Bl et | aoe eon onen® || Sas 2S 
Sa Sica | emai en | rau s 
e Sill HE | See MNO os aT = Date. Food. Remarks. 
o -= on) o o's feb) =) ara a 5 aS 
[om = no} a Oo SOE a 2 as 
Pe se Sy ROR eer Sh Ore & 
Feel Ores |e eueron | oles 
1 | 290 | 3-719 eee 
2] 215 | 4615 | 
915 5 
|e 7 oe merit -4°593 15 52 Jan. 30 Melox Room is heated 
| 5 | 140 5°635 
6 | 170 4°42] 
1, | 290 3691 |: 
2/215 | 4584 | 
‘ 4°] , z Z i : 
II. . : cae ee -4°568 15 52 Feb. 6 Melox Room is heated 
| 5 | 135 5°594 | 
6 | 165 4488 | J 
1 | 295 | 3-783 | | 
2} 210 4°657 5 
TIL. 4] 3 | 245 | 4-216 | -4°595 | 15 | 42 | Mar.9 Meoxs {| Ge 
4 145 4-779 | \ the operation 
5 | 135 5°544 
112995 | 3-075 ‘,Rusks and lean | Eleventh day of 
91990 | 3868 | meat rusks and lean 
IV.4|3| 245 | 3:957 | }4077 | 15 | 42 | Mar. 21; pps oot 
4|145 | 4-799 Forty-two days 
5 135 4-692 after the opera- 
. : tion 
2| 905 | 92-765 Rusks and thyroid | Sixth day of thy- 
SONS lso09 | = s:10004 : roid feeding. 
V.4} 41130 | 4-446 | (2728 15 45 | Mar. 30- Fifty-one days 
| 5 | 190 | 4583 | | | after the opera- 
tion 
fs Rusks and_ para- | Seventh day of 
3 | 210 | 3347 | thyroid parathyroid 
Wie | 4.) 112, |) 4:946 ( 4192 15 48 Apr. 7 - teeding 
5 | 112 | 4285 | ) | Sixty days after 
| the operation 


Studies on the Endocrine Glands 367 


TasLe VIII.—CO, Excretion. 
Group C (Thyroidectomised Rats, Male). 


| 


i . o- ° | 
_ Ee — : I j 
iw!) . | -=.08] 3S | ee | & 
S 18| 4 | sem| 35) 25 | # | 
7) - oP “|. 6. na = a Tae = 
Bicol ec leegeleoae|os| & 
“> 3 = | ge | Ol | be g Date. Food. Remarks. 
. a » ig = | ) 
BP igi = | S83 | P42 188] < 
Reem jon | Saco \|c.| g 
A“ Orets | &.9.9 | a & 3 
“455 /4%| & 
1/195 | 4-682 | °F. | 1916 
2|180| 4647 | 
ax . 
IL : 5 or 4°590 15 52 Jan. 31 Melox Room is heated 
5 | 180 4°689 | 
6 | 160; 4710 a QO (ec ee 
eT a RS NT a (a a a a BT ! 
1 | 195 4619 |) 
2| 180; 4598 
Me aro) aco, |p 4575 | 15 | 5a | Feb. 7 Melox Room is heated 
5 | 180| 4652 See) | | | fl . 
6 | 165 4°709 
1/195 | 3°774 | 
=) 460; 3°816 |{ : Thirty days after 
anf 3]$B| ao fs fe | ee | sae ate 3 | 950 | 3-540 | 3°703 15 42 Mar. 10 Melox { the operation 
4 | 165 | 3684 
( 1} 195 | 3:352 h Rusks and _ lean | Eleventh day of 
IV. }| 2] 185 | 3°513 |} 3-257 15 42 | Mar. 21 1 meat rusks and lean 
\ 3 | 250} 2-908 J meat feeding ! 
Foy. | Seventh day of 
w4| 2 oo otal 3°110 15 45 | Mar. 31 | Rusks + thyroid thyroid feed- 
3 | 225 | 3351 ing? 
| Rusks + parathy- | Seventh day of 
Vi 3 | 240 | 3504 | 3504 15 48 Apr. 7 roid parathyroid 
feeding * 


' Forty-one days after the operation. 
* Fifty-one days after the operation. 
3 Fifty-nine days after the operation. 


VOL. XI., NOS, 3 AND 4.—1917. 24, 


368 


TaBLE IX.—CO, EXcRETION OF 


Kojima 


THREE MALE RATS UNDER VARYING CONDITIONS 
OF FEEDING. 


| 


added to the rusks 


Sus | 8 55, Sg F 
. ~ . Tela Of Sites = 
225 |°o.|) aie Pe ayeelbercr ea ene 3 
= B : is is a 5 as o a = Date. Food. Remarks. 
a tet eee ec se. |S 
Spot EL ese Be) We eee, a) lee = 
SI 3 a © ZS ol Boa Oo nei S| 
A ara. ee Cae sl 
5 aH ay) <i 4 jee) | 
alte 1916. 
( 1 | 155 3°633 ] 
I.¢ | 2] 150] 4-668 4216 15 48 May 3 | Rusks, 
| 3 | 280 | 4°338 | 
(| From May 12-18 | Third day of para- 
{ 1) 160") 935624277) | O'lgrm,ofdryox-| thyroid feeding 
II.< | 2 | 145 | 4-660 4:27] 15 5] May 14- parathyroid per 
( 3 | 220] 4511 | rat per diem was 
| \| added to the rusks 
From May 12-18 | Sixth day of para- 
( 1 | 165 | 3°750 \ O-l grm.ofdryox-| thyroid feeding 
TII.; | 2 | 140 | 4-483 4°22] 15 52 May 17 - parathyroid per 
\ 3 | 225 | 4-432 | | rat per diem was 
\| added to the rusks 
1 | 165 3°683 | ) 
Ve 2} 145 | 4:496 | + 4-200 15 52 May 24 | Rusks 
3) (2879), <4:zonual) 
From May 26-June |Fifth day of thy- 
1 | 150 | 2965 | ) | 5, 3grm. of fresh | roid feeding 
Vie 2) 1380] 4-033 | > 3:724 15 54 May 30 - sheep-thyroid per 
3 | 210 | 4174 | | | | rat per diem was 
| | added to the rusks 
| From May 26—June | Tenth day of thy- 
( 1 | 140 | 3°160 | 5, 3 grm. of fresh roid feeding 
Vas Qe 125" al |) 4-02 | 15 54 June 4- sheep-thyroid per 
| 3 | 200 4°493 | rat per diem was 
| 


Studies on the Endocrine Glands 369 


TaBLe X.—CO, Excretion or Taree FemaLe Rats UNDER VARYING CONDITIONS 


OF FEEDING. 


and VI. 


Sele | S.=| 352 | 23| 2 
2 S a) 2 20 5 ct a 2 
f= 2 mB me — Pa ond 2. 
ret © i ges | Ole | BE, Date. Food. Remarks, 
S12) 5) %8o!/ e738 |42/ 8 
BES | oae | £23 / 52] = 
Bip | AF |} Otn |] SBo] oe S 
= Nw | Sag |/2e| 3 
7 aa 
*F 1916 
( 4/180) 3-741 | | 
I. | 51140] 3-900 | -3-762 | 15 | 48 | May 3 | Rusks 
\ 6} 185 | 3-645 | | | 
| 
|Rusks and para- | Thirdday of para- 
| 4/175 | 3°794 | | thyroid the same |_ thyroid feeding 
II.+| 5 | 146 | 3-864 | /3:747 15 51 May 14 as above table, 
| 6 | 187 | 3585 | Experiments II. 
and III. 
Rusks and _ para- | Sixthday of para- 
| 4/175 | 3682 | thyroid the same | thyroid feeding 
4am 5 | 143 3°851 -3°708 15 52 May 17 as above table, 
| 6 | 185 | 3592 | Experiments I1. 
and III. 
{| 4} 175) 3571 | | 
IV.+| 5 | 150 | 3-946 | ;3-746 15 52 May 24 | Rusks 
|} 6| 190 | 3-723 | | 
Rusks and _ fresh} Fifth day of thy- 
| 4/167} 2:989 | sheep-thyroid as| roid feeding 
1 V.5) 5 | 145 3121 -3'079 15 54 May 30 in Table IX., 
\| 6 | 130 | 3127 | Experiments V. 
and VI. 
( | Rusks and _ fresh | Tenth day of thy- 
| 4] 160} 3-172 | | sheep-thyroid as| roid feeding 
Wi. 5 | 140 3°835 | -3°5U7 15 o4 June 4- in Table IX., 
; | 6| 170] 3515 


| | Experiments V. 


370 Kojima 
Taste XI.—CO, Excretion or ContRots (Matz). 
oor | S. D 
. 43 © A eis i, & Pm | oe = 
& Sia seme Meee as pervusy  lhy Slee || age 
SP eligeii est pes Speere een) oe 
5 Po 2 Dea |(Royescee Pies =i Date. Food. Remarks. 
Be Be ee Scena ae. (ae eae AS 
a fa a S26 | S&S | So NS 
m Sadan towers ll asea ose leeieci tl ica Im 
Be Weeleoaes | sl ste ll aaa e 
4 S) moO > ti “a oO [o) 
<4 gq aa} 
2 rele 1916. 
CNTOT I TSOnl eA 1OI Iq) ee sees Es 
fe 108 | 150 | 4382 | J 4:241 15 49 | May 4] Rusks 
107 | 185 | 4-253 |) ,.; 3 P a 
I. { 7 | ase || aie |ff2t7 | 28 | SL | May 15 | Rusks 
fal LOT 1808) p4s195. eye 2 by : 
J 108 | 155 | 4172 [f 4183 15 52 | May 18 | Rusks 
(| 107 | 180 | 4-219 |) : : ey, 
IV. {} 108 | 155) 4-151 | J 4185 15 53 | May 25 | Rusks 
(leors| diss) | 4-905" ))_ 4. p “ 
V. LU] 108 | 155 173 |J 4189 15 54. | May 31] Rusks 
LOM alpeliS: 4°211 z p . 
Wil \ 108 i fae } 4199 15 55 | June 5 | Rusks 


Studies on the Endocrine Glands 371 


Taste XII.—CO, Excretion or Four Castratep Rats (MALE). 


bh ve | =. d 
s |. | 23.,|S8m| Pe) § | | 
mye |e | get eanrlpesl| 3 
=| * = 2°45 3. 2 es = 
2 ms a os 0 sz = 47 & 
: 2 | S's oy, 2, / 
FE A d Ee & Oss e's = Date. Food. | Remarks. 
2i1/2/5)ee5] e838 | 52] 8 | 
5 ale | wee | fee pee) Ss 
al ee = O 5s oA ° e bs 5 
=. > oo 4 se = 
_ 72 ee 
. 
110 | 245 | 4:317 F. 1916. 
111 | 270 | 4109 , 
7 : rere +) r 14] Rusks 
112 | 275 | 4-438 Neaigs 15 O | May 1 ake | 


113 | 285 4°602 


110 | 245 3°265 
111 | 290 | 3°234 | 
112 | 300 | 3-573 | 
113 | 300 | 2°586 


| | ‘Twenty-second 
-3°164 15 54 | June 7| Rusks-| day after cas- 
tration 


| Forty - eighth 
day after cas- 
| tration 


110 | 240} 3143 
111 | 300 3181 


Taste XIII.—CO, Excretion or Four Entire Rats (MALE). 


be Dig tik a 
Mice | a.) £8. |-825| 221 § 
Peer |} sa lace | 3,8) ee | s 
FI = os) O's oO ap = so 
=| ° co) o4 D oO iS) ee =| = 
‘ © . oar (Ooo) Be Sg Date. | Food. Remarks, 
m4 = = Sum | Pa | FS 
n” 5 S aes x a Oo so S 
=| 22 Peo | Hs | 8 
4 4°93 /49| @ 
114 | 250 | 4-296 ] F. 
I l a oe per ia 15 | 52 | May 15| Rusks 


4°325 15 


54 e 8 | Rusks 
c 4| Rusks 


| 
4°304 15 56 


372 


CuHartT 1,—Amount of food consumed by rats per kilo of body-weight and per week in grams. 


Kojima 


400 


BG 


Week 


ending 


Apr. 


, controls ; B, parathyroidectomised ; C, thyroidectomised ; M.=meat ; R.=rusks. 
The animals of B and C groups were operated on in the second week. From the first to the 
sixth week they were fed with melox; during the seventh and eighth weeks with meat and 


rusks ; during the ninth week with rusks and thyroid, and during the tenth week with rusks 
and parathyroid. 


Cuanrr 2,—Total N in food consumed per kilo of body-weight and per week in grams. 


I Il V IX he 
ww ~ = a a a is > 
o.5 
SS oS n fa 

i=] - - ) a a x « fo¥ 
ez 3 : os 2 : : oe Mae 


A, controls ; B, parathyroidectomised ; C, thyroidectomised. 


The animals of B and C groups were operated on in the second week. From the first to the 
sixth week they were fed with melox; during the seventh and eighth weeks with meat and 
rusks ; during the ninth week with rusks and thyroid, and during the tenth week with rusks 
and parathyroid. 


374 Kojima 


Cuart 3.—Total N in urine per kilo of body-weight and per week in grams. 


; ia 

; = 
ae 

: ci 
ae 


Hi 


Z 


I Itt LV". VE Vio Vil Vil. TX ae 
go z = 2 e S = Z en 
SS os i = 
FE = = s = 2 = = = st 


A, controls ; B, parathyroidectomised ; C, thyroidectomised. 
The animals of B and C groups were operated on in the second week. From the first to the 
sixth week the animals of all the groups were fed with melox only ; during the seventh and eighth 


weeks with meat and rusks; after the ninth week with rusks, with the addition of thyroid and 
parathyroid respectively in consecutive weeks. 


Studies on the Endocrine Glands 375 


Cuart 4.—Total N retained in body per kilo of body-weight and per week in grams. 


J II I IV V VI Vi Vill Ix x 

to ~ Lan) o re) oo oOo ™ boa) al ™~ 
rs P=) _ a N re — N oD 

5:5 « .s “ 

es 2 ‘ . - 3 ; : ‘ 3 A 

a ss io 5 é = ‘i rs is < 


A, controls; B, parathyroidectomised ; C, thyroidectomised. 

B and C groups were operated on in the second week. From the first to the sixth week the 
animals of all the Soe were fed with melox ; from the seventh to the eighth weeks with lean 
meat and rusks, and during the ninth week with rusks with addition of thyroid. During the tenth 
week they were fed with rusks, with addition of parathyroid. 


Cuart 5,—Total Ca in food consumed per kilo of body-weight and per week in grams. 


Il VI VIL Vii TX 
we 0 a a = oo = 3 a m3 = 
o 
aS as s i : ‘ i = 
Be 2 : : = = ; ; : <q 


A, controls ; B, parathyroidectomised ; C, thyroidectomised. 

The animals of B and C groups were operated on in the second week. From the first to the 
sixth week they were fed with melox; during the seventh and eighth weeks with meat and 
tusks; during the ninth week with rusks and thyroid, and during the tenth week with rusks 
and parathyroid, 


376 Kojima 


Cuarvr 6,—Total Ca in urine per kilo of body-weight and per week in grams. 
0-003 


~ 
Ss 
Ss 
> 
Q 


I Il il IV V VI Vin VIE IX XS 


Week 
ending 


”) 
LB) 
? 
” 
99 


Apr. 


A, controls ; B, parathyroidectomised ; C, thyroidectomised. 


The animals of B and C groups were operated on in the second week, From the first to the 
sixth week they were fed with melox ; during the seventh and eighth weeks with meat and 


rusks ; during the ninth week with rusks and thyroid, and during “the tenth week with rusks 
and parathyroid, 


Cuarr 7.—Total Ca retained in body per kilo of body-weight and per week in grams. 


Week 
ending 
Feb. 


A, controls ; B, parathyroidectomised ; C, thyroidectomised. 


The animals of B and C groups were operated on in the second week. From the first to the 
sixth week they were fed with melox; during the seventh and eighth weeks with meat and 


rusks ; during the ninth week with rusks aud- thyroid, and during the tenth week with rusks 
and parat hyroid. 


Studies on the Endocrine Glands 377 


Cuart 8,—Amount of food consumed by male and female rats per kilo of 
body-weight and per week in grams. 


550 


500 


950 


400 


350 
wet = n a 
a3 Ob z 
eo & = : =} 
es = ar 


M.=male; F.=female. 
May 12-18: Parathyroid added to rusk diet. 
May 26-June 1: Thyroid added to rusk diet. 


CHART 9.—Total N in food consumed per kilo of body-weight 
and per week in grams. 


a II It [V 
we 5 = at a 
SS 2 
a er : ‘ 5 


M.=male; F.=female. 


May 12-18: Parathyroid added to rusk diet. 
May 26-June 1: Thyroid added to rusk diet. 


378 Kojima 


Cuarr 10,—Total N in urine per kilo of body-weight 
and per week in grams. 


M.=male; F.=female. 
May 12-18: Parathyroid added to rusk diet. , 
May 26-June 1: Thyroid added to rusk diet. 


Cuarr 11.—Total N retained in body per kilo of body-weight 
and per week in grams, 


Fen Sea 


ney 
o.5 
Ory 
we 
>| 
(cota) 


M.=male; F. =female. 


May 12-18: Parathyroid added to rusk diet. 
May 26-June 1: Thyroid added to rusk diet. 


Studies on the Endocrine Glands 379 


Cuart 12.—Amount of CO, excretion per hour and per kilo in grams, 


1 2 3 4 5 6 


A, controls; B, parathyroidectomised ; C, thyroidectomised. 


A group: 1, 2, and 3 show the amount of CO, during melox feeding, while 4 shows the amount 
of CO, on the tenth day of meat and rusk feeding; 5, the amount of CO, on the seventh day 
of thyroid feeding ; 6, that on the seventh day of parathyroid feeding (see Table VI.). 

B group: 1 and 2 show the amount of CO, before parathyroidectomy during melox feeding ; 
3, thirty days after the operation, also fed with melox ; 4 shows the amount of CO, forty-two days 
after the operation and on the eleventh day of meat and rusk feeding; 5, the amount on the 
sixth day of thyroid and rusk feeding and fifty-one days after the operation ; 6, the amount on the 
seventh day of parathyroid and rusk feeding and sixty days after the operation (see Table VII.). 

C group: land 2 show the amount of CO, before thyroidectomy during melox feeding ; 3, thirty 
days after the operation, also fed with melox ; 4, forty-one days after the operation and on the 
eleventh day of meat and rusk feeding ; 5, fifty-one days after the operation and on the seventh 
day of thyroid and rusk feeding ; 6, fifty-nine days after the operation and on the seventh day of 
parathyroid and rusk feeding (see Table VIII.). 


380 Studies on the Endocrine Glands 


CHARr 13.—Amount of CO, excretion in grams. 


M.=male rats; F.=female rats; C.=control male rats. 


1, all the animals fed with rusks alone; 2, CO, estimated on third day of parathyroid and 
rusk feeding; 3, CO, estimated on sixth day of parathyroid and rusk feeding ; 4, CO, estimated 
during rusk feeding; 5 and 6, CO, estimated on fifth and tenth days respectively of thyroid and 
rusk feeding. The control rats were fed throughout with rusks alone (see Tables IX., X., 
and XI.). 


CHART 14.—Amount of CO, excretion in grams. 


SY 


a b c 


Ca. = castrated rats ; N. =normal rats. 
a, CO, estimated before castration ; b, CO, estimation twenty-two days after 
castration ; c, CO, estimation forty-eight days after castration. 


A CROSS-STRIATED MAMMALIAN MUSCLE PREPARATION. By 
R. J. S. M‘Dowati.! (From the Department of Physiology, 
Edinburgh University.) (With nine figures in the text.) 


ORDINARY mammalian muscle is disadvantageous for experimental work, 
since it dies soon after removal from the body unless kept at about body 
temperature and perfused with oxygenated blood or other suitable fluid. 

Although the retractor penis of the hedgehog, which consists of plain 
muscle, is known to live for a long time after removal from the body and 
without perfusion, it does not seem to be generally known that certain 
cross-striated muscles of the same animal retain their functions equally 
well under these circumstances, and are otherwise not unsuitable for 
experimentation under ordinary laboratory conditions. 

The muscles in question are those concerned with the rolling of the 
animal into a ball and the subsequent unrolling. These muscles and their 
actions are described by Huxley in his Anatomy of the Vertebrata. 
They are composed, as already stated, of cross-striated fibres, although it 
must be stated that in some of them appearances are seen which suggest 
incomplete histogenesis. 

They fall into two groups, according to their form and attachments. 
One, the orbicularis dorsalis, a very thick circular band of fibres, plays 
the chief part in the curling up of the animal. It is, however, so closely 
incorporated with the skin that it cannot readily be dissected off without 
lacerating its fibres. The other group, which is concerned in starting both 
the rolling and the unrolling process, is formed of straight, parallel- 
fibred muscles which arise from the trunk and are inserted into the skin. 
There is no difficulty in dissecting them out in their whole length (5-7 cm.) 
without damage. Removed from the body and kept at ordinary laboratory 
temperature, without perfusion or special oxygenation, they continue excit- 
able for at least twenty-four hours. They contract on stimulation at all 
temperatures, from 0° to 40°C. 

In the experiments shown in the illustrations accompanying this paper 
the Keith-Lucas method of stimulation was employed. By this method 
the whole diameter of a muscle is made to complete a circuit between two 
electrolytic solutions: thus all the fibres of the muscle are stimulated and 


1 The observations embodied in this paper were made four years ago. It was intended 
to pursue the investigation further in the following year, but the war prevented this inten- 
tion being carried out ; and as the author is still on military service, it has been thought 
well not to delay publication any longer.—[{Ep. ] 


382 M‘Dowall 


the polarising action of metal electrodes is avoided. The apparatus was 
arranged so that it could be immersed in a vessel of water and raised or 


; 


Fic, 1.—Diagram of apparatus employed. 


A, B, muscle chamber in two parts, which fit together exactly ; C, holein the bottom of 
part A, through which one end of the muscle strip, E, is passed: A and B are filled 
with Ringer’s solution ; D, D’, platinum electrodes in the Ringer ; F, thermometer 
indicating the temperature of the fluid; G, muscle lever ; K, K, outer vessels form- 
ing a water jacket to the muscle chamber; H, H, inlet tube for passing either 
warmed or cooled water into the water jacket; I, I, siphon for drawing water off; 
L, lamp; M, gutta-percha covered wire passing through water jacket. 

By the above arrangement any current passing from D to D’ must traverse thelwhole 
thickness of the muscle. 


lowered in temperature at will, the temperature being recorded at about 
I cm. distance from the muscle (fig. 1). The following points have been 
made out in the course of these experiments :— 


Preparation 


A Cross-Striated Mammalian Muscle 


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VOL. XI., NOS. 3 AND 4. 


384. M‘Dowall 


1. If the muscles are kept stretched they retain their excitability for 
a much longer time than when unweighted. 


xdxexfxgx 


Fic. 4.—Ordinates showing the extent of contraction in response to single 
induction shocks of varying intensity. 


a, 600 Kronecker units e, 4000 Kronecker units 
b, 1000 i f, 5000 . 

c, 2000 as g, 6000 = 

d, 3000 5 


Fig. 5.—Curves showing the extent of contraction in response to rapidly repeated stimuli 


of varying intensity. The numbers below each curve indicate the numbers of units 
on a Kronecker coil. 


2. Muscles which have been removed from the body remain excitable 
longer than those which are left in situ. 


3. On stimulating the weighted muscle, it passes fairly quickly (after 


A Cross-Striated Mammalian Muscle Preparation 


Fic. 6.—Curves illustrating the effect of varying temperatures on the hedgehog muscle. 


385 


Fic. 7.—Fatigue curve of hedgehog muscle, taken at room temperature. 


386 M‘Dowall 


a latent period of about ‘04 second) into the contracted condition; but 
relaxation occurs only very gradually and occupies a long time (fig. 2). 

4. In consequence of the length of the relaxation period, the summation 
of stimuli can be produced and tetanus obtained by excitations which recur 
at unusually long intervals (fig. 3). 

5. Within limits the amount of contraction is proportional to the 


Fic, 8.—Fatigue curve from a muscle which hed been already completely fatigued and then 
subjected to a prolonged period of rest. The curve on the right is taken after the muscle had 
been more heavily weighted. (At the beginning of the experiment the stimuli were more 
frequent than afterwards. ) 


Fic. 9.—Comparison curves from a muscle which had not been previously fatigued. 
On the left unweighted, on the right weighted. 


strength of the stimulation (fig. 4 for single excitations and fig. 5 for 
multiple excitations). On tetanisation with strong currents the muscle 
contracts to as much as one-fourth of its original length. 

6. As with other muscles, the periods both of contraction and of 
relaxation are shortened by heat and lengthened by cold (fig. 6). 

7. The preparation lends itself well to the investigation of the effects 
of fatigue (tig. 7), and especially to demonstration of the fact that muscular 
fatigue may be recovered from, even in the mammalian preparation, with- 
out the necessity for any call upon the circulating fluid. Thus a muscle 


A Cross-Striated Mammalian Muscle Preparation 387 


which had been completely fatigued gave, after a prolonged rest, a series 
of contractions (fig. 8) very little if at all inferior to those yielded by the 
similar muscle of the opposite side, which had not been stimulated (fig. 9). 

Incidentally it may be noted that the tracings on the right of figs. 8 
and 9 show that the contracture which is usually described as characteristic 
of fatigue is not seen if the muscle be sufficiently weighted. 


I have to thank Dr John Tait for suggesting the search for a sur- 
viving muscle-preparation in the hedgehog. 


BINDING SECT. JUL 1 2 1967 


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