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STUDY OF THE DISTRIBUTION OF
IODINE BETWEEN CELLS AND COL-
LOID IN THE THYROID GLAND
A DISSERTATION
SUBMITTED TO THE FACULTY
OF THE OGDEN GRADUATE SCHOOL OF SCIENCE
IN CANDIDACY FOR THE DEGREE OF
DOCTOR OF PHILOSOPHY
DEPARTMENT OF PHYSIOLOGICAL CHEMISTRY AND PHARMACOLOGY
BY
HARRY BENJAMIN VAN DYKE
Private Edition, Distributed By
THE UNIVERSITY OF CHICAGO LIBRARIES
CHICAGO, ILLINOIS
Reprinted from
'he JOURNAL OF BIOLOGICAL CHEMISTRY, Vol. XLV, No. 2, 1921 ; Vol. LIV, No. i, 1922
The AMERICAN JOURNAL OF PHYSIOLOGY, Vol. LVI, No. i, 1921
tlntversitg'of 'Gbicago
A STUDY OF THE DISTRIBUTION OF
IODINE BETWEEN CELLS AND COL-
LOID IN THE THYROID GLAND
A DISSERTATION
SUBMITTED TO THE FACULTY
OF THE OGDEN GRADUATE SCHOOL OF SCIENCE
IN CANDIDACY FOR THE DEGREE OF
DOCTOR OF PHILOSOPHY
DEPARTMENT OF PHYSIOLOGICAL CHEMISTRY AND PHARMACOLOGY
BY
HARRY BENJAMIN VAN DYKE
Private Edition, Distributed By
THE UNIVERSITY OF CHICAGO LIBRARIES
CHICAGO, ILLINOIS
Reprinted from
The JOURNAL OF BIOLOGICAL CHEMISTRY, Vol. XLV, No. 2, 1921; Vol. LIV, No. i, 1922
The AMERICAN JOURNAL OF PHYSIOLOGY, Vol. LVI, No. i, 1921
BfOLOGY
LIBRARY
G
CXCHANGE
. ; &
Reprinted from THE JOURNAL OF BIOLOGICAL CHEZJ^TRY, Vol. XLV, No. 2, 1921
A STUDY OF THE DISTRIBUTION OF IODINE BETWEEN
CELLS AND COLLOID IN THE THYROID GLAND.
II. RESULTS OF STUDY OF DOG AND HUMAN THYROID GLANDS.
BY HARRY BENJAMIN VAN DYKE.
(From the Laboratory of Physiological Chemistry and Pharmacology,
. University of Chicago, Chicago.}
(Received for publication, November 29, 1920.)
In the first paper of this series Tatum1 described a method
whereby thyroid cells may be separated from colloid material
and examined chemically. Briefly the method consists in cutting
frozen sections of the thyroid gland and floating these sections
on Ringer's2 solution. The colloid material immediately drops
out of the acini and is presumably dissolved in the Ringer's solu-
tion. The cells may then be separated by centrifugalization,
dried, weighed, and analyzed. Comparisons between the iodine
content of cells so separated and the iodine content of control
pieces of unsectioned whole gland may indicate the distribution
of iodine between cells and colloid under different functional
conditions.
In this paper I wish to report the results of a study, suggested
by Dr. A. L. Tatum, of the distribution of iodine in the thyroid
glands of normal and iodine-fed dogs as well as in human glands
obtained from individuals subjected to operation for toxic goiter.
The method earlier described by Kendall3 was used in making the
final iodine determinations.
Incidental to the determination of the iodine distribution in
dog and human thyroid glands, some control experiments were
performed relative to the alterability of the intracellular iodine
concentration during the process of cutting. It may be argued
that a portion of the iodine-containing compound diffuses from
1 Tatum, A. L., J. BioL Chem., 1920, xlii, 47.
2 Campbell, J. A., Quart. J. Exp. Physiol., 1911, iv, 1, Formula "A."
3 Kendall, E. C., J. BioL Chem., 1914, xix, 251.
325
THE JOURNAL OF BIOLOGICAL CHEMISTRY, VOL. XLV NO. 2
520421
326 Distribution of Ibdme -In. Thyroid Gland. II
cells as they lie suspended in Ringer's solution during the sepa-
ration of the colloid material. If it is assumed that such an
outward diffusion takes place, it seems reasonable to expect that
the iodine compound should diffuse back into the cells if we should
increase the concentration of that iodine compound in the Ringer's
solution in which the cells are suspended. Several experiments
like those given in Table I were undertaken to determine whether
or not the iodine content of the cells could thus be increased.
TABLE I.
Effect of Floating Cells on Pure Ringer's Solution and on Ringer's Solution
on Which Cells of an Iodine-Rich Gland Had Been Floated.
Animal
No.
Weight
of
whole
gland
used.
Iodine
in
whole
gland.
Weight
of
cell
mass
used.
Iodine
in
cell
mass.
Ratio of
percentage of
iodine in cells
to percentage of
iodine in
whole gland.
Remarks.
mg.
percent
mg.
percent
14
146.0
0.187
40.5
0.017
0.091
Cells floated on pure Ring-
er's solution.
53.0
0.017
0.091
Cells floated on Ringer's
solution on which previ-
ously cells of iodine-rich
gland had been floated.
20
287.2
0.030
161.5
0.005
0.167
Cells floated on pure Ring-
er's solution.
151.0
0.006
0.20
Cells floated on Ringer's
solution on which previ-
ously cells of iodine-rich
gland had been floated.
It may be seen from the data given in Table I that no increase
in the iodine content of cells could be brought about by increas-
ing the concentration of the characteristic iodine compound in
the Ringer's solution.
Again the iodine concentration of the cell mass does not seem
to be in the least altered whether or not the freshly centrifugalizecl
cell mass is washed several times with iodine-free Ringer's solution.
Moreover, in the attempt to find a suspending medium more
nearly related physicochemically with thyroid cells than is Rin-
ger's solution, I have used fresh dog serum and have not found
the iodine content of the cells measurably different from that of
control cells suspended in Ringer's solution.
H. B. van Dyke
327
Finally autolysis does not seem to be much of a factor in the
loss — if any occurs — of iodine compounds from cells as they lie
in contact with Ringer's solution. Throughout the process of
cutting and separating the cells, the reaction of the suspending
medium should remain weakly alkaline — a reaction which has
been shown to be unfavorable to autolysis.4 Also in experiments
to be reported later in which the same technique was used under
slightly different conditions, every effort was made to eliminate
autolysis by cutting successively small portions of the gland and
floating the cells on Ringer's solution cooled by ice. The cells
from the small portions cut successively were at once centrifu-
galized and dried. No change could be noticed in the iodine
distribution.
Results of Study of Dog Glands Taken at Random.
TABLE II.
Quantitative Determination of Iodine in Whole Gland and in Cells Free from
Colloid Material of Thyroid Glands of Normal Dogs.
Animal
No.
Weight
of whole
gland
used.
Iodine
in whole
gland.
Weight
of cell
mass
used.
Iodine
in cell
mass.
Ratio of
percentage
of iodine in
cells to
percentage
of iodine in
Morphology.
Cells.
Colloid.
whole gland.
mg.
per cent
mg.
per cent
1
5
550.0
0.25
154.0
0.031
0.124
Flat.
Fair.
6
388.0
0.175
265.0
0.027
0.154
ft
Rich.
10
480.0
0.017
342.0
0.003
0.176
Cuboidal.
Poor.
11
505.5
0.032
197.5
0.005
0.156
u
"
12*
278.7
0.006
286.5
13
689.0
0.234
154.0
0.047
0.201
Flat.
Fair.
14
146.0
0.187
40.5
0.017
0.091
u
ft
19
474.5
0.024
116.0
"
Poor.
20*
287.2
0.030
161.5
0.005
0.167
21*
604.7
0.038
189.0
0.006
0.158
23
458.2
0.003
390.4
Cuboidal.
Poor.
24
318.7
0.021
295.4
"
"
31
513.8
0.055
205.6
0.008
0.145
tt
"
32*
235.2
0.179
103.9
0.037
0.207
33
289.3
0.011
538.3
0.002
0.182
Flat.
Poor.
* No histological examination.
4 Bradley, H. C., J. BioL Chem., 1915, xxii, 113. Bradley, H. C., and
Taylor, J., /. BioL Chem., 1916, xxv, 261.
328 Distribution of Iodine in Thyroid Gland. II
From the results given in Table II one sees that the ratio of
iodine concentration in cells to iodine concentration in whole
gland (and hence the ratio of iodine concentration in cells to
colloid-iodine concentration) has a quite constant value. In
some glands (Nos. 12, 19, 23, and 24), to be sure, the iodine con-
tent of the cell mass analyzed was so low that no ratio value
could be obtained. But in general, despite great variations in
the iodine content and morphology of the glands analyzed, the
ratio values change relatively little. These findings in the dog's
thyroid gland are similar to those of Tatum1 in the beef, pig, and
sheep thyroid glands. A comparison of the ratio values in dif-
ferent animals is given below.
Effect of Feeding Iodine.
It was thought that perhaps ratio changes could be induced by
the administration of iodine or iodine compounds. Capsules
containing 1 gm. of potassium iodide or 2 drops of tincture of
iodine in starch were fed over varying periods of time. No more
than one capsule was given in 24 hours. The results of the
examination of the glands of animals so treated are given in
Table III.
The ratio obtained from analyzing the cells and whole gland of
the thyroid glands of animals which had received varying amounts
of iodine or iodine compounds over periods of no less than 3 days
is practically the same as that of the animals which had received
no iodine. One gland (that of No. 16) gave an unusually high
value which I cannot explain. In view of the variations in mor-
phology and iodine content of the glands of the normal series
together with the quite constant ratio exhibited by that series,
it is not surprising that the ratio still remains constant despite
the feeding and consequent absorption of iodine. From a con-
sideration of the glands examined it appears that the ratio is not
altered by the feeding of iodine as potassium iodide or free iodine
over periods of time ranging from 3 days to 3 weeks; and yet
the total iodine content of these glands undoubtedly is greatly
increased during that same period.
H. B. van Dyke
329
TABLE III.
Quantitative Determination of Iodine in Whole Gland and in Cells Free from
Colloid Material of Thyroid Glands of Iodine-Fed Dogs.
Period of feeding.
Ratio of
percent-
!Ani-
mal
No.
Form of iodine.
Daily.
On
alter-
nate
Total.
Weight
of
whole
gland
used.
Iodine
in
whole
gland.
Weight
of
cell
mass
used.
Iodine
in
cell
mass.
age of
iodine in
cells to
percent-
age of
iodine in
days.*
whole
gland.
days
days
days
mg.
per cent
mg.
per cent
2
KI
14
0
14
98.2
0.31
77.7
0.045
0.145
7
KI
14
7
21
147.0
0.12
113.0
0.031
0.258
8
KI
14
7
21
124.5
0.87
125.0
0.080
0.092
9
KI
14
2
16
353.5
0.251
116.0
0.040
0.159
16
KI
12
0
12
651.4
0.319
316.1
0.117
0.367
17
KI
Tincture of I.
12
12
0
0
24
646.5
0.617
302.3
0.083
0.134
27
(( it T
3
0
3
556.1
0.302
332.8
0.046
0.152
29
l( (( T
3
0
3
195.2
0.314
158.6
0.039
0.124
30
l< (( T
3
0
3
163.3
0.357
127.5
0.041
0.115
After feeding daily.
Human Glands.
The distribution ratio of iodine between cells and colloid mate-
rial was determined in thirteen human glands obtained from
operative cases.5 In Table IV are given the results of the anal-
yses of the human glands together with the clinical diagnosis
made in connection with ten of the cases.
It will be seen that most of the thyroid glands reported were
clinically diagnosed as toxic goiters. Considerable variations in
the ratio value occur and do not appear to be related either to the
iodine content of the gland or to its morphology. The ratio
value of No. 22 is inexplicably high. In the human gland
series, however, as in other gland series previously reported, the
ratio variations are of a much smaller magnitude than the varia-
tions in total iodine content. So here too the ratio is fairly
constant despite variations in morphology and iodine content.
6 Through the courtesy of Dr. C. B. Davis and Dr. A. D. Bevan of the
Presbyterian Hospital, Chicago, and of Dr. A. J. Ochsner of Augustana
Hospital, Chicago.
330 Distribution of Iodine in Thyroid Gland. II
TABLE IV.
Quantitative Determination of Iodine in Whole Gland and in Cells Free from
Colloid Material of Human Thyroid Glands.
Series No.
Weight of whole
gland used.
b
*0
X!
1
a
'i!
•g-a
Weight of cell
mass used.
1
a
II
3s
Ratio of percent-
age of iodine in
cells to per-
centage of io-
dine in whole
gland.
Morphology.
Diagnosis.
Cells.
Colloid.
mg.
per
cent
mg.
per cent
22
284.7
0.081
260.8
0.049
0.605
Cuboidal.
Poor.
Colloid cystic
goiter with
toxic symp-
toms.
25*
600.2
0.274
308.6
0.105
0.383
Flat.
Rich.
Mild exophthal-
411.0
0.276
240.7
0.098
0.355
mic goiter.
35
671.8
0.286
179.2
0.051
0.178
Flat.
Rich.
Toxic thyroid.
36
439.7
0.158
219.9
0.020
0.127
Cuboidal.
Fair.
" goiter.
37f
667.5
0.092
258.4
0.027
0.293
Colloid goiter.
38
578.6
0.079
191.7
Trace.
Cuboidal.
Poor.
« «
following par-
enchymatous
hyperplasiaof
exophthalmic
goiter.
39f
516.0
0.279
297.6
0.095
0.341
40
469.2
0.345
225.5
0.066
0.191
Flat.
Rich.
Exophthalmic
goiter.
41
559.2
0.280
187.0
0.067
0.239
(i
«
48*
549.9
0.043
312.4
0.005
0.116
Cuboidal.
Poor.
607.3
0.044
311.3
0.005
0.114
49*
590.7
0.137
256.0
0.026
0.189
Cuboidal.
Fair.
Exophthalmic
204.5
0.027
0.197
goiter.
51
731.4
0.283
294.4
0.072
0.254
Flat.
Rich.
« «
60
370.0
0.152
228.0
0.015
0.099
it
«
Cystic goiter
with toxic
symptoms.
* Duplicate determinations made,
t No histological examination.
H. B. van Dyke
331
Comparison of Ratio Value in Different Animals.
A comparison of the ratio values of the thyroid glands of the
different animals so far examined is given in Table V. The few
abnormally high and unexplained ratio values are not included in
the table.
From what data are available there appear to be some differ-
ences in the thyroid glands of different animals in the numerical
value of the ratio of the percentage of iodine in cells to that in
whole gland. The ratio value of iodine distribution for dog
thyroid glands seems to be consistently lower and more constant
than that for the thyroid glands of the other animals studied.
TABLE v.
A Comparison of the Value of the Ratio of the Percentage of Iodine in Cells
Free from Colloid Material to the Percentage of Iodine in Whole
Gland in Different Animals.
Animal.
Extremes of
iodine content
of whole gland.
Ratio values.
Extremes.
Mean.
Average.
Beef* ...
per cent
0.023-0.468
0.011-0.870
0.043-0.345
0.377-0.810
0.089-0.442
0.21 -0.48
0.091-0.258
0.099-0.384
0.20 -0.34
0.23 -0.41
0.35
0.175
0.242
0.27
0.32
0.36
0.154
0.22
0.27
0.33
Doe
Man
Pig*f
Sheep*
* Tatum.1
t Only two glands analyzed.
SUMMAKY.
1. The method described by Tatum1 was used to determine
the ratio of the percentage of iodine in cells to the percentage of
iodine in whole gland in the thyroid glands of normal and iodine-
fed dogs as well as in human thyroid glands obtained from opera-
tive cases.
2. Evidence is presented indicating that the concentration of
intracellular iodine is independent of the suspending medium,
whether that is pure Ringer's solution, Ringer's solution containing
iodine-rich colloid material, or homologous blood serum.
332 Distribution of Iodine in Thyroid Gland. II
3. The ratio value was found to be relatively constant despite
great variations in the morphology and iodine content of the
glands examined. The ratio value for the dog's thyroid gland
seems quite constant and is much lower than that of the beef or
sheep.
Reprinted from THE AMERICAN JOURNAL OF PHYSIOLOGY
Vol. 56, No. 1, May, 1921
A STUDY OF THE DISTRIBUTION OF IODINE BETWEEN
CELLS AND COLLOID IN THE THYROID GLAND
III. THE EFFECT OF STIMULATION OF THE VAGO-SYMPATHETIC NERVE
ON THE DISTRIBUTION AND CONCENTRATION OF IODINE IN THE
DOG'S THYROID GLAND
HARRY BENJAMIN VAN DYKE
From the Laboratory of Physiological Chemistry and Pharmacology, University of
Chicago
Received for publication February 5, 1921
For many years it has been held that the thyroid gland is supplied
with true secretory nerves. In support of this assertion there is con-
siderable anatomical evidence and some physiological evidence. Of
late the nerves which anatomists have traced into the thyroid gland and
have considered to be possibly secretory in function have been declared
to be branches of the cervical sympathetic nerve. And recent physi-
ological work has tended to confirm this view. Only twelve years ago
Wiener (1) published the report of experiments from which he concluded
that extirpation of the inferior cervical ganglion produces a marked
atrophy of the thyroid gland on the side of the extirpation. Wiener
maintained that no comparable effect on the lobe of the thyroid gland
on the side of the operation could be produced by vagotomy or by
removal of the superior cervical ganglion. More recently Rahe et al. (2)
announced that they were able to produce a quite marked diminution
in the iodine concentration of the lobe of the thyroid gland on one side
by stimulating the thyroid nerves in several different ways. They
stimulated the nerves of the superior thyroid artery, the intact vago-
sympathetic nerve as well as the vago-sympathetic nerve near the level
of the superior cervical ganglion after ligating the nerve low in the neck
and cutting the nerve central to the point of stimulation. They found
that the most marked loss was brought about by the stimulation of the
intact vago-sympathetic nerve.
Watts (3) undertook to find out whether or not the results obtained
by Rahe, Rogers, Fawcett and Beebe might be due to vasomotor
168
DISTRIBUTION OF IODINE IN THYROID GLAND 169
changes in the gland on the stimulated side. Watts likewise found
that he could reduce the iodine content of the right or left lobe of the
thyroid gland of the dog by stimulating the "cervical sympathetic iso-
lated from the vagus sheath" and the "nerve filaments accompanying
the superior thyroid vessels." However Watts maintained that he
could cause some diminution in the iodine content simply by periodi-
cally reducing the blood flow through the gland by "occluding the main
thyroid artery" the nerves of which had been dissected away. Hence
he concluded that all of the effects of stimulation on the iodine content
can be accounted for by the coincident vasomotor changes which he
showed to be present.
Positive evidence of the secretory effect of sympathetic stimulation
has been reported by Cannon and his co-workers (4) in several communi-
cations. Working with cats they sutured the phrenic nerve with the
cervical sympathetic nerve and observed following the operation in-
creased basal metabolism, respiratory hippus and falling hair which
they interpreted as the results of hypersecretion of the thyroid gland
caused by the periodic bombardment of the gland by impulses carried
from the respiratory center to the gland's secretory nerves. Cannon
and Cattell (5) adduced additional evidence as to the role of the sympa-
thetic nerves in experiments dealing with the electrical condition of the
gland. Following the stimulation of the upper thoracic sympathetic
nerves or the injection of epinephrin they were able to show a definite
action current in the thyroid gland after a latent period of five to seven
seconds. Recently Cannon and Smith (6) maintained that gentle mas-
sage of the thyroid gland or stimulation of the cervical sympathetic
nerve increases the rate of the denervated heart. The denervated
heart is said not to be affected when the cervical sympathetic nerve is
stimulated after removal of the thyroid gland. Moreover Levy (7) ob-
served that the pressor effect of epinephrin after a variable latent period
is increased by the stimulation of the cervical sympathetic nerve. He
declared that stimulation of the cervical sympathetic nerve has no such
effect after thyroidectomy.
However the conclusions based on the experiments mentioned above
have not been universally accepted. Burget (8) was unable to alter the
thyroid gland noticeably either by uniting the phrenic and cervical
sympathetic nerves or by removing a section of the cervical sympathetic
nerve. Marine, Rogoff and Stewart (9) sutured together the phrenic
and cervical sympathetic nerves in several cats. They demonstrated a
functional union between the phrenic and .cervical sympathetic nerves
170 HARRY BENJAMIN VAN DYKE
but observed no exophthalmos, tachycardia or respiratory hippus in
their animals. There was no apparent difference either grossly or his-
tologically in the lobes of the thyroid gland on the operated and the non-
operated sides. Troell (10) reported that he was unable to produce
either exophthalmos or respiratory hippus by suturing the proximal end
of the phrenic nerve to the cervical sympathetic nerve. Employing
cocaine as a sensitizer for sympathetic nerve endings, Mills (11) did
not observe, following the repeated injection of cocaine, any alteration
in the amount or nature of the thyroid secretion as judged by what is
known of the gland's histology. Finally Rogoff (12) records one experi-
ment in which he drew blood from a vein of the left lobe of the thyroid
gland and at the same time stimulated the cervical sympathetic nerve
on that side in the hope of increasing the secretory activity of the stimu-
lated lobe. From the right lobe he also collected blood by way of a
vein. While drawing the blood he massaged the right lobe to some
extent but did not stimulate the right cervical sympathetic nerve. He
found that specimens of dried blood from each lobe were potent when
fed to tadpoles. But he could detect iodine chemically only in the
blood from the non-stimulated lobe. Moreover the non-stimulated lobe
had a lower iodine content than the stimulated lobe.
In connection with some studies on the distribution of iodine in cells
and colloid in the thyroid gland I attempted to alter acutely the total
iodine content of the gland by stimulating the vago-sympathetic nerve
of the dog. Some inconsistencies in the results in the early part of
the work forced me to investigate the matter more carefully and to
repeat the work of Rahe et al. (2) and of Watts (3).
Methods. Dogs were used in all of the experiments. All except
those whose numbers are above that of no. 78 were given daily feedings
of iodine over a period of one to eleven days. The daily feeding con-
sisted of a capsule containing two drops of tincture of iodine in starch.
In the animals fed iodine the stimulation of the vago-sympathetic nerve
was undertaken from two to ten days after the last feeding of iodine.
Throughout the experiments the animals were lightly anesthetized with
ether. Platinum wire electrodes were applied to opposite sides of the
carefully isolated vago-sympathetic nerve and shielded from all sur-
rounding tissues by glass. In all of the experiments except those re-
corded in table 5, a tetanizing current from three to six times as strong
as that sufficient to bring about a pupillary dilatation and apparent
protrusion of the bulbus oculi was employed. The regulation of the
strength of the current was made possible by a rheostat inserted in the
DISTRIBUTION OF IODINE IN THYROID GLAND
171
circuit. Again in all of the experiments except those to be found in
table 5 the stimulating current throughout the period of stimulation
was made for about 0.8 second at intervals of 1.6 seconds by means of
a clock and ratchet device. The strength of the current used and the
rate at which the current was made in the experiments of table 5 are
described below.
TABLE i
Variations in the iodine content of neighboring specimens of the same lobe of the
thyroid gland
ANIMAL NUMBER
NUMBER OF
FEEDINGS OF
IODINE
LOBE OF
THYROID GLAND
WEIGHT OF SAM-
PLE OF DRIED
GLAND ANALYZED
IODINE IN DRIED
GLAND ANALYZED
DIFFERENCES IN
THE CONCENTRA-
TION OF IODINE
IN NEIGHBORING
SPECIMENS OF
THE SAME LOBE
gram
per cent
per cent
Left
0.1192
0.1378
0.331
0.424
0.093
68
11
Right I
0.1525
0.1668
0.418
0.455
0.037
/
/
0.6931
0.366
0.005
Left
0.6420
0.371
0.034
I
0.6846
0.405
0.039
74
6
(
0.4549
0.370
0.006
Right \
0.4931
0.364
0.019
1
0.5533
0.383
0.013
Left
0.4454
0.5298
0.121
0.132
0.011
88
0
•
Right |
0.7857
0.4528
0.119
0.129
0.010
Left
0.7241
0.7138
0.158
0.176
0.018
105
0
Right |
0.4489
0.4516
0.163
0.148
0.015
After a number of experiments had been performed it was found that
there is considerable variation in the iodine content of adjoining pieces
of the same gland. In table 1, for example, are given a few analyses
of neighboring specimens of the same gland. The differences in neigh-
boring portions of the glands are somewhat greater in iodine-fed animals.
172
HARRY BENJAMIN VAN DYKE
TABLE 2
Quantitative determination of iodine in whole gland and in cells free from colloid
material of thyroid glands of dogs whose isolated vago-sympathetic nerve had
previously been stimulated for approximately three hours
ANIMAL NUMBER
LOBE
STIMU-
LATED
LENGTH OF PERIOD
OF STIMULATION
WEIGHT OF
WHOLE GLAND
USED
IODINE IN WHOLE GLAND
WEIGHT OF CELL MASS
USED
IODINE IN CELL MASS
RATIO OF PER CENT OF
IODINE IN CELLS TO
PER CENT OF IODINE
IN WHOLE GLAND
APPARENT GAIN (+) OK
LOSS ( — ) IN CONCEN-
TRATION OF IODINE IN
WHOLE GLANU OF STIM-
ULATED LOBE
gram
per cent
gram
per cent
per cent
42
Left
2 hrs. 15 min. <
L 0.1792
R 0.1693
0.531
0.560
0.2533
0.1360
0.371
0.303
0.699
0.541
-0.029
45
Left
3 hrs. 0 min. <
L 0.4602
R 0.5336
0.148
0.102
0.5117
0.4750
0.011
0.014
0.074
0.137
+0.046
46
"Left
3 hrs. 30 min. <J
L 0.4982
R 0.5252
0.098
0.104
0.3445
0.2284
0.014
0.017
0.143
0.163
-O.OC6
54
Right
2 hrs. 30 min. <
L 0.3964
R 0.4241
0.177
0.147
0.2713
0.3883
O.C36
0.019
0.203
0.129
-0.030
55
Right
2 hrs. 40 min. <
L 0.6880
R 0.5509
0.232
0.186
0.2700
0.2280
0.031
0.046
0.134
0.247
-0.046
56
Right
2 hrs. 10 min. <
L 0.4536
R 0.4795
0.121
0.109
0.3683
0.3059
0.016
0.014
0.132
0.128
-0.012
62
Left
3 hrs. 5 min. <
L 0.0836
R 0.0558
0.494
0.523
0.0480
O.C612
0.078
0.082
0.158
0.157
-0.029
64
Right
3 hrs. 50 min. <
L 0.0892
R 0.0692
0.110
0.127
0.1019
0.1190
0.008
0.010
0.073
0.079
+0.017
65
Right
3 hrs. 0 min. <
L 0.1179
R 0.1419
0.436
0.466
0.1086
0.1064
0.037
0.052
0.085
0.112
+0.030
66
Right
3 hrs. 0 min. <
L 0.0709
R 0.0593
0.485
0.493
0.0733
0.0783
0.041
0.038
0.085
0.077
+0.008
67
Left
3 hrs. 0 min. <
L 0.1373
R 0.1237
0.464
0.451
0.1842
0.1613
0.041
0.040
0.088
0.089
+0.013
68
Left
2 hrs. 45 min. <
L 0.2570
R 0.3193
0.377
0.437
0.1703
0.1379
0.031
0.028
0.082
0.064
-0.060
69
Left
3 hrs. 0 min. <
L 0.5360
R 0.3957
0.605
0.569
0.2627
0.1476
0.083
0.092
0.137
0.162
+0.036
71
Right
3 hrs. 0 min. <
L 0.3651
R 0.2990
0.346
0.305
0.1718
0.1310
0.026
0.029
0.075
O.C95
-0.041
DISTRIBUTION OF IODINE IN THYROID GLAND 173
Hence it was thought desirable to analyze not single blocks or sam-
ples of dried powdered mixtures of whole gland but to analyze the whole
gland in each case. In experiments in which this last mentioned tech-
nique was employed the whole gland was carefully cleaned of connec-
tive tissue and blood vessels, and thoroughly dried first over an electric
hot plate and then in an electric oven. After two to three hours' dry-
ing in the electric oven the gland was broken into several pieces whose
weight was about 0.5 gram each and whose number therefore depended
on the size of the gland. The iodine determinations were made accord-
ing to the method earlier described by Kendall (13). By analysis of
powdered thyroid of known iodine content the accuracy of the method
(to about 0.008 mgm. of iodine) and the purity of the reagents used
were frequently examined and found to be satisfactory.
The strength of the current employed in all of the experiments re-
corded in table 2 was three times that sufficent to cause dilatation of
the pupil and apparent protrusion of the bulbus oculi. Both the stimu-
lated and the non-stimulated vago-sympathetic nerves were cut in
two places: at a point in the neighborhood of the eighteenth tracheal
ring and also at a level a little above that of the hyoid bone. The reason
for cutting the non-stimulated nerve in such a manner was to eliminate
the possible effect of tonic secretory impulses on the non-stimulated
lobe. The vago-sympathetic nerve was stimulated a little above the
point at which it was cut low in the neck. In the above experiments
the ratio of the percentage of iodine in cells to the percentage of iodine
in whole gland was determined by a method previously described (14).
From the data given in table 2 it may be seen that stimulation of the
vago-sympathetic nerve under the conditions described is without ap-
preciable effect on either the ratio value or the concentration of iodine
in the whole gland.
When it was found that there was no consistent diminution in the
concentration of iodine in the stimulated lobe only the stimulated
vago-sympathetic nerve was sectioned in the manner described above.
In experiments 76, 79 and 80 a strength of current six times that neces-
sary to cause ocular changes characteristic of sympathetic stimulation
was used; in all of the other experiments to be found in table 3 the cur-
rent was of the same strength as that used in the experiments recorded
in table 2. From the standpoint of the iodine concentration in whole
gland the results given in tables 2 and 3 are very similar. Stimulation
apparently has no effect on the concentration of iodine in the stimulated
lobe.
174
HARRY BENJAMIN VAN DYKE
By stimulating the intact vago-sympathetic nerve Rahe et al. (2) de-
clare that they were able to produce the most marked diminution in the
iodine content of a given lobe of the thyroid gland. In the three experi-
ments of table 4 of my series, the intact vago-sympathetic nerve was
TABLE 3
The concentration of iodine in the lobes of the thyroid gland of the dog after the
stimulation of the isolated vago-sympathetic nerve on one side for a period of
approximately three hours
ANIMAL
NUMBER
LOBE STIMU-
LATED
LENGTH OF PERIOD OF
STIMULATION
WEIGHT OF
DRIED WHOLE
GLAND
IODINE IN
DRIED WHOLE
GLAND
APPARENT GAIN
(+)ORLOSS(-)
IN CONCENTRA-
TION OF IODINE
IN STIMULATED
LOBE
gram
per cent
per cent
72
Right
2 hrs. 50 min. <
L 1.2883
R 1.4258
0.186
0.189
+0.003
74
Right
3 hrs. 10 min. <
L 2.0197
R 1.5013
0.381
0.373
-0.008
75*
Left
3 hrs. 10 min.
L 1.4975
R 1.8796
0.334
0.360
-0.026
76
Right
3 hrs. 10 min. |
L 0.4653
R 0.3509
0.770
0.821
+0.051
77
Right
3 hrs. 30 min. <
L 0.7055
R 0.7914
0.471
0.508
+0.037
79
Right
3 hrs. 15 min. <
L 0.2421
R 0.1995
0.474
0.498
+0.024
80
Left
4 hrs. 0 min. <
L 0.6342
R 0.4850
0.465
0.477
-0.012
82
Left
3 hrs. 10 min. <
L 0.8483
R 0.7516
0.203
0.206
-0.003
83
Left
3 hrs. 15 min. \
L 1.2435
R 0.9163
0.105
0.103
+0.002
84
Left
3 hrs. 10 min. <
L 0.7582
R 0.6022
0.129
0.145
-0.016
Entire right lobe not analyzed.
DISTRIBUTION OF IODINE IN THYROID GLAND
175
stimulated in the neck at about the level of the fifteenth tracheal ring.
The current was of the same strength as that employed in the experi-
ments of table 2; each time the current was made there ensued a respira-
tory arrest and the ocular changes typical of sympathetic stimulation.
Again there was no consistent change in the concentration of iodine in
the stimulated lobe.
TABLE 4
The concentration of iodine in the lobes of the thyroid gland of the dog after the
stimulation of the intact v ago-sympathetic nerve on one side for a period of approx-
imately three hours
ANIMAL
NUMBER
LOBE STIMU-
LATED
LENGTH OP PERIOD OP
STIMULATION
WEIGHT OF
DRIED WHOLE
GLAND
IODINE IN
DRIED WHOLE
GLAND
APPARENT GAIN
(,+) OR LOSS (— )
IN CONCENTRA-
TION OF IODINE
IN STIMULATED
LOBE
gram
per cent
per cent
78
Left
3 hrs. 40 min. <
L 0.4628
R 0.3706
0.528
0.501
+0.027
86
Right
3 hrs. 20 min. <
L 0.7039
R 0.6919
0.138
0.125
-0.013
88
Left
3 hrs. 15 min. <
L 0:9752
R 1.2385
0.127
0.123
+0.004
Effect of vasomotor activity on the concentration of iodine in the thyroid
gland. It will be recalled that Watts (3) concluded from his experi-
ments that vascular changes will account for the diminution in the con-
centration of iodine which he brought about by stimulation of the cer-
vical sympathetic nerve. The experiments of table 5 were undertaken
to find out whether or not a slightly different type of stimulus sent into
the nerve at an interval more nearly like that employed by Watts had
an effect comparable to that found in the experiments previously per-
formed. It was also thought desirable to determine whether or not
the characteristic vascular changes were present throughout the
experiments.
The technique except for certain features of the stimulation was the
same as that used in all of the preceding experiments. Usually the
stimulated vago-sympathetic nerve was ligated and cut at about the
level of the eighteenth to twentieth tracheal ring; near the ganglion
nodosum only the vagus nerve was cut. In all of the experiments the
sympathetic chain was intact above the eighteenth tracheal ring. The
TABLE 5
The concentration of iodine in the lobes of the thryoid gland of the dog after the stim-
ulation of the v ago-sympathetic nerve, the sympathetic portion of which was left
intact above the point of stimulation
ANIMAL
NUM-
BER
LOBE STIMU-
LATED
LENGTH OF PERIOD OF
STIMULATION
PRESENCE
(+) OR AB-
SENCE ( — )
OF VA8O-
MOTOR AC-
TIVITY AT
END OF EX-
PERIMENT
WEIGHT OF
DRIED WHOLE
GLAND
IODINE IN
DRIED
WHOLE
GLAND
APPARENT GAIN
(+)ORLO8S ( — )
IN CONCENTRA-
TION OF IODINE
IN STIMULATED
LOBE
grams
per cent
per cent
101
Right
3 hrs. 10 min.
- {
L 1.8213
R 1.5890
0.177
0.125
-0.052
102
Left
3 hrs. 0 min.
+ {
L 0.1397
R 0.1727
0.141
0.120
+0.021
103
Right
3 hrs. 0 min.
+ {
L 0.6335
R 0.5749
0.277
0.274
-0.003
104
Right
3 hrs. 0 min.
- {
L 0.1891
R 0.1844
0.190
0.164
-0.026
105
Right
3 hrs. 25 min.
+ {
L 1.4379
R 0.9005
0.167
0.156
-0.011
106
Left
3 hrs. 25 min.
+ {
L 0.6161
R 0.6624
0.043
0.041
+0.002
107
Right
3 hrs. 0 min.
- {
L 3.1065
R 2.9642
0.103
0.109
+0.006
108
Left
3 hrs. 35 min.
+ {
L 0.3462
R 0.2754
0.021
0.027
-0.006
109
Right
3 hrs. 25 min.
* {
L 1.0407
R 1.0116
0.014
0.016
+0.002
111
Left
3 hrs. 50 min.
+ {
L 0.2875
R 0.2638
0.076
0.075
+0.001
113
Right
3 hrs. 30 min.
- {
L 1.5649
R 2.6585
0.034
0.033
-0.001
114
Right
3 hrs. 30 min.
+ {
L 0.3210
R 0.3690
0.143
0.132
-0.011
115
Left
3 hrs. 45 min.
+ {
L 1.0381
R 0.8196
0.088
0.090
-0.002
116
Left .
3 hrs. 30 min.
- {
L 0.6650
R 0.6753
0.016
0.017
-0.001
117
Left
4 hrs. 20 min.
+ {
L 1.1050
R 1.1701
0.140
0.134
+0.006
118
Right
5 hrs. 0 min.
- {
L 0.6568
R 0.5036
0.456
0.438
-0.018
119
Right
3 hrs. 10 min.
{
L 0.6866
R 0.5695
0.168
0.173
+0.005
176
DISTRIBUTION OF IODINE IN THYROID GLAND 177
vago-sympathetic nerve was stimulated just above the point at which
it was cut near the eighteenth tracheal ring. It was easily possible to
vary the strength of the electrical stimulus so that a current of such a
strength was employed as just to bring about the ocular changes char-
acteristically associated with stimulation of the cervical sympathetic
nerve. Mendenhall (15) has emphasized the markedly toxic effects of
ether on the sympathetic nervous system. In a preparation like that
last described the sensitivity of the cervical sympathetic nerve to the
depressant action of ether could easily be demonstrated. A tetanizing
current of rather low frequency from a Stoelting inductorium was made
for 5.5 seconds at intervals of 11.8 seconds throughout the period of
stimulation. At the end of most of these experiments a vein of the
stimulated lobe, in most cases without the ligation of its companion
veins, was cannulated and the effect on blood flow of stimulation of the
gland under the same conditions as those employed in the experiment
was observed. In ten out of sixteen experiments no difficulty was en-
countered in demonstrating a vasoconstriction in the gland on stim-
ulating the vago-sympathetic nerve with an electric current of the same
strength and delivered at the same rate as that used in the previous
stimulation period. The threshold of excitability for the vasoconstrictor
nerves of the thyroid gland appears to be considerably lower than that
for the submaxillary gland as reported by Gruber (16). The relatively
low threshold of excitability of the vasoconstrictor nerves of the thyroid
to epinephrin stimulation has been observed by Gunning (17).
From table 5 it may be seen that stimulation of the vago-sympathetic
nerve with the sympathetic trunk intact above the eighteenth tracheal
ring has no appreciable effect on the concentration "of iodine in the stim-
ulated lobe. In a number of experiments it was unequivocally demon-
strated that vasoconstrictor fibers to the gland were being stimulated;
yet such stimulation did not alter detectably the iodine content of the
lobe subjected to stimulation.
Effect of stimulation of the vago-sympathetic nerve on the concentration
of water in the thyroid gland. In table 6 may be found data relative to
the effect of stimulation of the vago-sympathetic nerve on the concen-
tration of water in the thyroid gland. I was unable to find that stimu-
lation had any effect on the concentration of water in the stimulated
lobe even in experiments in which vasomotor changes were definitely
proved to result from stimulation.
178
HARRY BENJAMIN VAN DYKE
TABLE 6
The concentration of water in the lobes of the thyroid gland of the dog after the stimu-
lation of the vago-sympathetic nerve for a period of from three to four
hours
ANIMAL
NUMBER
LOBE
STIMULATED
LENGTH OF PERIOD OP
STIMULATION
CONCENTRATION OF
WATER
APPARENT GAIN (+)
OR LOSS ( — ) IN CON-
CENTRATION OF
WATER IN STIMU-
LATED LOBE
per cent
per cent
77
Right
3 hrs. 30 min. <
L 69.25
R 69.64
+0.39 .
79
Right
3 hrs. 15 min. <
L 73.40
R 74.26
+0.86
82
Left
3 hrs. 10 min. <
L 74.06
R 74.66
-0.60
83
Left
3 hrs. 15 min. <
L 77.57
R 78.02
-0.45
84
Left
3 hrs. 10 min. <
L 74.38
R 74.24
+0.14
88
Left
3 hrs. 15 min. <
L 73.65
R 72.81
+0.84
108*
Left
3 hrs. 35 min. <
L 80.14
R 80.71
-0.57
109*
Right
3 hrs. 25 min. <
L 78.54
R 80.64
+2.10
110*
Right
3 hrs. 45 min. <
L 82.69
R 82.13
-0.56
111*
Left
3 hrs. 50 min. <
L 78.03
R 77.33
+0.70
113
Right
3 hrs. 30 min. <
L 75.98
R 73.73
-2.25
115*
Left
3 hrs. 45 min. \
L 75.76
R 75.63
+0.13
116
Left
3 hrs. 30 min. <
L 79.92
R 78.34
+1.58
117*
Left
4 hrs. 20 min. <
L 76.53
R 75.65
+0.88
119
Right
3 hrs. 10 min.
L 75.90
R 76.81
+0.91
* Vasomotor effect of the stimulation demonstrated at the end of the experi-
ment.
DISTRIBUTION OF IODINE IN THYROID GLAND
179
DISCUSSION
If we consider the data described above we find that following the
stimulation of a given lobe of the thyroid gland there was no consistent
change in the concentration of iodine in that lobe compared with the
TABLE 7
A comparison of the effect of the different methods of stimulation of the vago-sympa-
thetic nerve on the concentration of iodine in the thyroid gland of the dog
SERIES
NON-
IGNIFI-
CANT
APPARENT LOSS IN CON-
CENTRATION OF IODINE
APPARENT GAIN IN CON-
CENTRATION OF IODINE
REMARKS
Num-
ber
Maxi-
mum
Aver-
age
Num-
ber
Maxi-
mum
Aver-
age
per cent
per cent
per cent
per cent
I
2
7
0.060
0.035
5
0.046
0.028
Stimulated vago-sympa-
thetic nerve cut above
hyoid bone and low in
the neck. Non-stimu-
lated vago-sympathetic
nerve cut similarly in
most cases
II
4
3
0.026
0.018
3
0.051
0.037
Only stimulated vago-
sympathetic nerve cut
as in series I
III
1
1
0.013
0.013
1
0.027
0.027
Intact vago-sympathetic
nerve stimulated at
about level of fifteenth
tracheal ring
IV
11
5
0.052
0.024
1
0.021
0.021
Vago-sympathetic nerve
stimulated at about
level of eighteenth tra-
cheal ring. Sympathet-
ic nerve entirely intact
above point of stimula-
tion
V
7
2
0.011
0.011
1
0.021
0.021
Animals of series IV in
which vasomotor effect
•
of stimulation could be
shown plainly at end of
experiment
nonstimulated lobe. Following stimulation there was an apparent
diminution in the concentration of iodine in 24 or 54.5 per cent of the
stimulated glands, and an apparent gain in 20 or 45.5 per cent of glands
stimulated similarly. Watts (3) reported that the average difference
180 HARRY BENJAMIN VAN DYKE
in the percentage of iodine in the lobes of dried thyroid gland of the dog
is 0.015 per cent in this vicinity. If we consider as non-significant all
differences in the percentage of iodine in dried thyroid of 0.010 per cent
or less we see that stimulation, while causing an apparent diminution
in the concentration of iodine in 16 or 36.4 per cent of the glands, and
an apparent increase in the concentration of iodine in 10 or 22.7 per
cent of the glands, had no effect on the iodine concentration of 18 or
40.9 per cent of the stimulated glands. The greater part of the experi-
mental data presented may be briefly summarized in table 7.
The differences which I found above appear to be due to normal
variations in the concentration of iodine in the two lobes of the dog's
thyroid gland. The average percentage difference in the concentration
of iodine in the two lobes of the dog's thyroid gland depends on a num-
ber of factors such as the type of gland, the time of year and the feed-
ing of iodine. Hence I am forced to conclude that if stimulation of the
vago-sympathetic nerve in the dog has any effect on the concentration
of iodine in the thyroid gland, that effect is considerably less than the
normal variation in the iodine content of the stimulated and control
lobes. The presence of normal variations relatively so much greater
than the variations which may follow stimulation renders valueless the
application of more refined methods of iodine determination in the study
of the effect of stimulation of the cervical sympathetic nerve on the
concentration of iodine in the thyroid gland.
SUMMARY
1. Periodic stimulation of the isolated vago-sympathetic nerve by
an induced current of a moderate to strong intensity over a period of
from three to three and a half hours does not appreciably alter the
distribution ratio of iodine between cells and colloid.
2. The findings of Rahe, Rogers, Fawcett and Beebe (2) and of Watts
(3) that stimulation of the cervical sympathetic nerve for a comparable
period of time reduces the concentration of iodine in the stimulated
lobe were not confirmed. Watts' assertion that such stimulation reduces
the water content of the stimulated lobe was not confirmed.
3. Conclusions as to the direct secretory control which the cervical
sympathetic nerves exercise on the thyroid gland are based in no small
measure on the alleged effect of stimulation of the cervical sympathetic
nerve on the iodine content of the gland. Conclusions having such a
basis apparently are untenable.
DISTRIBUTION OF IODINE IN THYROID GLAND 181
It is a pleasure to acknowledge the many suggestions and constant
aid given me by Dr. A. L. Tatum during the progress of this work.
BIBLIOGRAPHY
(1) WIENER: Arch, exper. Path. u. Pharm., 1909, Ixi, 297.
(2) RAHE, ROGERS, FAWCETT AND BEEBE: This Journal, 1914, xxxiv, 72.
(3) WATTS: This Journal, 1915, xxxviii, 356.
(4) CANNON, BINGER AND FITZ: This Journal, 1915, xxxvi, 363.
CANNON AND FITZ: This Journal, 1916, xl, 126.
(5) CANNON AND CATTELL: This Journal, 1916, xli, 58, 74.
(6) CANNON AND SMITH: Endocrin., 1920, iv, 386.
(7) LEVY: This Journal, 1916, xli, 492.
(8) BURGET: This Journal, 1917, xliv, 492.
(9) MARINE, ROGOFF AND STEWART: This Journal, 1918, xlv, 268.
(10) TROELL: Arch. Int. Med., 1916, xvii, 382.
(11) MILLS: This Journal, 1919, 1, 174.
(12) ROGOFF: Journ. Pharm. Exper. Therap., 1918, xii, 193.
(13) KENDALL: Journ. Biol. Chem., 1914, xix, 251.
(14) TATUM: Journ. Biol. Chem., 1920, xlii, 47.
(15) MENDENHALL: This Journal, 1914, xxvi, 57.
(16) GRUBER: This Journal, 1915, xxxvii, 259.
(17) GUNNING: This Journal, 1917, xliv, 215.
Reprinted from THE JOURNAL OF BIOLOGICAL CHEMISTRY
Vol. LIV, No. 1, September, 1922
A STUDY OF THE DISTRIBUTION OF IODINE BETWEEN
CELLS AND COLLOID IN THE THYROID GLAND.
IV. THE DISTRIBUTION OF IODINE IN THE HYPERPLASTIC THY-
ROID GLAND OF THE DOG AFTER THE INTRAVENOUS
INJECTION OF IODINE COMPOUNDS.
BY HARRY BENJAMIN VAN DYKE.
(From the Laboratories of Pharmacology, University of Chicago, Chicago.)
(Received for publication, July 20, 1922.)
Some years ago Marine and Feiss (1) and Marine and Hogoff
(2, 3) first performed experiments which leave little doubt as to the
ability of the dog's thyroid gland, especially when hyperplastic,
to bind iodine almost instantaneously. Marine and Feiss (1)
carefully perfused the surviving thyroid gland with fluid con-
taining iodine as KI. They found that after 1 hour a consider-
able amount of iodine was taken up only by a surviving gland.
Any evidence of death of the perfused organ was accompanied by
a loss of some of the gland's stored iodine rather than by an
absorption of iodine from the circulating medium. However, even
a surviving gland, rich in iodine, lost some iodine to a perfusing
fluid free of the element. In surviving glands they discovered
that the amounts of iodine absorbed were relatively independent
of the amounts of iodine in the perfusing fluids. They also pointed
out that a similarly rapid absorption of iodine by the intact gland
follows the intravenous administration of a solution of KI. From
the results of perfusions of spleen and kidney under similar condi-
tions they concluded that these organs were not capable of taking
up a significant amount of iodine.
Marine and Rogoff (2) on the basis of experiments in which
they injected a solution of KI intravenously came to the con-
clusion that the absorption of iodine by the gland is almost as great
1 hour after the injection as it is 30 hours after the injection.
They again found that no significant amount of iodine was taken up
by the spleen and liver. The amount of iodine absorbed by the
11
12 Iodine in the Thyroid Gland
thyroid gland appeared to be directly proportional to the degree
of hyperplasia exhibited by the gland. In a second communica-
tion (3) they discussed the time of appearance of the changes in the
histology and physiological activity of the gland following the
intravenous administration of KI solution.
Having found (4) that there was relatively little difference in
the ratio of the percentage of iodine in cells to the percentage of
iodine in whole gland in dog thyroid glands exhibiting great
variations in histological appearance and iodine content, I under-
took the present study to determine what effect acute iodization of
hyper plastic thyroid glands has on the ratio value.
Methods.
As in the work of Marine and Feiss, and Marine and Rogoff,
dogs with thyroid glands usually definitely hyperplastic were
used in all of the experiments. Light ether narcosis was always
employed. All solutions of KI and thyroid colloid were injected
into the femoral vein. Every effort was made to section the glands
as rapidly as possible after their removal from the animal. In
the making of the determinations of the ratio of the percentage
of iodine in dried cells to that in dried whole gland the method
first described by Tatum (5) was somewhat modified. To lessen
autolysis as much as possible the glands on removal were plunged
into Ringer's solution cooled to 1^°C. The glands were then
carefully cut into blocks of a size appropriate for the floor of the
freezing microtome and frozen sections were made of a considerable
amount of tissue. The blocks of tissue cut for the microtome's
floor as well as the cut sections were suspended in Ringer's solution
cooled to 1-4°C. As in the preceding work of this series the frozen
sections were cut sufficiently thin so that practically all of the
colloid or intercellular fluid was dissolved out of the acini as soon
as the sections were suspended in cooled Ringer's solution. By
centrifugalization the cells were separated from the colloid dissolved
in the Ringer's solution and carefully dried over an electric hot-
plate and then in an electric drying oven. Control pieces of
uncut whole gland were also dried in the same manner. Through-
out this paper colloid iodine solutions refer to cell-free solutions of
thyroid intercellular fluids obtained by the method just described
and dissolved in Ringer's solution. In all of the experiments the
H. B. van Dyke 13
quantitative determinations of iodine were made by the method
of Kendall (6).
From the data of Table I it can be seen that large amounts of
iodine as KI were taken up by hyperplastic thyroid glands despite
big variations in the dose of KI (50 to 150 mg.) and duration of time
between injection and removal of gland (90 seconds to 22 hours
and 25 minutes). The cooling of the blocks of thyroid tissue dur-
ing the process of cutting had little effect. Either little of the
injected iodine was held by the cells, or it diffused rapidly from the
cells in the process of cutting; for the iodine content of the cells
and hence the ratio of the precentage of iodine in cells to the per-
centage of iodine in whole gland is considerably less than that
found in resting glands. The average ratio value of 0.15 for the
dog (4) was approached only in Experiment 11 in which nearly
24 hours elapsed between the injection of KI and the removal of
the experimental lobe. In this last case there are alternative
means of explanation: more iodine may have been bound by the
cells by removal from the thyroid's intercellular spaces, or the iodine
in the cells was in a less diffusible form than in the other experi-
ments of the series. These changes in the distribution of iodine
after about 24 hours can be definitely correlated with Marine and
Rogoffs (3) discovery that 20 hours after the injection of iodine as
KI, markedly hyperplastic glands exhibit more stainable colloid
with some increase in the size of the follicular spaces and shrinkage
in the height of the columnar epithelium.
In the experiments the data of which are given in Table II,
solutions of colloid iodine from normal dog thyroid glands were
injected intravenously to determine to what extent iodine in
organic combination is bound by hyperplastic thyroid glands.
The iodine of thyroid colloid of normal animals is practically
not at all taken up by hyperplastic glands after 42 to 86 minutes.
A comparable amount of iodine in the form of KI is rapidly
taken up.
Blum and Griitzner (7) found some hours after the injection
into the circulatory system of fluid pressed from thyroid glands
that the iodine-containing thyroid protein is split largely by
hepatic action into simpler products with the formation even
of iodides. In Experiments 12 and 13 (42 and 86 minutes after
intravenous injection) there is little evidence that much iodide-
TABLE I.
Ratio of the Percentage of Iodine in Cells to the Percentage of Iodine in Whole
Glands in Hyperplastic Thyroid Glands of Dogs Receiving an
Intravenous Injection of KI Solution for Varying
Lengths of Time before the Removal of the Glands.
Animal No.
p
H
*o
11
I.I
<! "
if8
m
|
°c -6
till
H
1
r
«*-T)
°8
I]
jg*
rS
13
^3
*
a
si
p
9
1
"8
11
3 3
£
Iodine in cell mass.
Ratio of per cent of
iodine in cells to
per cent of iodine
in whole gland.
mg.
sec.
gm.
per cent
gm.
per cent
1
50
90
Control.
0.6786
0.021
0.5242
0
Iodized.
0.5553
0.042
OT7845
0.002
0.048
2
50
150
Control.
0.6548
0.010
0.6373
0
Iodized.
0.5695
0.072
0.8437
0.003
0.042
tm'n.
3*
135
57
Control.
0.6797
0
0.3585
0
Iodized.
0.4808
0.086
0.5449
0
4
120
60
Control.
0.4512
0.042
0.3775
0
Iodized.
0.7760
0.079
0.4702
0
5
150
60
Control.
0.6853
0
0.6105
0
Iodized.
0.6302
0.078
0.7473
0
6
50
60
Control.
0.8779
0.002
0.5694
0
Iodized.
0.8314
0.059
0.8021
0.004
0.068
7
50
60
Control.
0.8130
0
Iodized.
0.652
0.085
0.6994
0.002
0.025
0.731
0.070
8
50
60
Control.
0.408
0.007
0.649
0.004
,
Iodized.
0.461
0.081
0.8222
0.004
0.045
0.245
0.098
9
150
64
Control.
0.515
0.002
0.7066
0
Iodized.
0.595
0.085
0.8282
0.003
0.035
10*
120
70
Control.
0.345
0.010
0.245S
0
Iodized.
0.250
0.091
0.2627
0
hrs. min
•
11
50
22 2
Control.
0.465
0.017
0.5357
0
0.571
0.020
Iodized.
0.719
0.213
0.5422
0.022
0.103
* Glands and cells not suspended in ice-cooled Ringer's solution.
14
H. B. van Dyke
15
iodine has been split from colloid iodine; for the hyperplastic
thyroid glands present exhibit no significant change in iodine con-
tent and yet are able quickly to bind any iodine available as iodide.
In the experiments reported in Table III colloid iodine solution
of animals with hyperplastic glands, each of which had received an
TABLE II.
Ratio of the Percentage of Iodine in Cells to the Percentage of Iodine in
Whole Gland in Hyperplastic Thyroid Glands of Dogs after the Intrav-
enous Injection of Thyroid Colloid Iodine from Normal Dog
Thyroid Glands. The Iodine Content of a Hyperplastic
Gland of a Dog after the Injection of a Comparable
Amount of Inorganic Iodine is also Recorded.
a
c
III
1
1
I**
1
o>
-^S^
•i
I
S
i
o||
|
1
0
11
t
1
•8
nt of iodin
elapsing
pletion of
removal
•8
s in whole t
t of cell m:
•
a
1
a
rtl
g
a
g
® a^Ja
0
'S'o
a
s
c
i 1
'3
<J ~"
. i
'4
H003"2
1
'Is
1
1
1
I'5"3
cc.
m0.
min.
am.
per
cent
am.
per
cent
12
200.0
Colloid
1.690
42
Control.
0.6565
0.002
0.5478
0.001
iodine.
Iodized.
0.6248
0.002
0.4819
0
13
259.6
Colloid
2.829
86
Control.
0.7971
0
0.7698
0
iodine.
0.8557
0
•
Iodized.
0.4742
0.005
0.9208
0
0.4245
0
14
241.5
KI
2.087
46
Control.
0.7939
0.003
0.6024
0.004
,.
Iodized.
0.4513
0.017
0.5907
0.019
intravenous injection of 50 mg. of KI 60 minutes before the removal
of the thyroid lobe for section, was injected intravenously as soon
as possible into other animals with hyperplastic glands. The
sequence was as follows: as a control, part of one lobe was removed;
colloid iodine dissolved in Ringer's solution was then injected and
all but half of one lobe resected; a comparable amount of iodine
as KI dissolved in a similar amount of Ringer's solution was then
16
Iodine in the Thyroid Gland
TABLE III.
Ratio of the Percentage of Iodine in Cells to the Percentage of Iodine in Whole
Gland in Hyperplastic Thyroid Glands of Dogs after the Intravenous
Injection of Thyroid Colloid Iodine from Hyperplastic Thyroid
Glands Iodized but a Short Time before Being Sectioned.
i
6*0 £
TJ
J.o
•**
—i £
•
•d
•s
§1 *.
•3
•d
3I'ST3
q
ii
_o
c
j
"o'c §
"3
'ui
•
•^ o "^b
6
ii
•gi
iil
ja
0
J
S
1
S
1
§ £.2
Sol
Animal K
it
ii
f
J*s * •
^u *&
I'll!
1
11
jjjj
'i
1
"8
f
Iodine in
Ratio of ]
in cells
£Ldine in
min.
gm.
per cenf
gm.
per
cent
15
0.820 mg. of col-
60
Control.
0.6022
Trace.
loid iodine of
Colloid
0.6986
0.009
1.1376
0
Animal 7* dis-
iodine.
0.7186
0.009
solved in 233 cc.
of Ringer's
solution.
0.836 mg. of
25
KI
0.8328
0.017
KI dissolved in
0.6935
0.022
253.5 cc. of
Ringer's solu-
tion.
16
0.908 mg. of col-
40
Control.
0.4700
0
loid iodine of
Colloid.
0.4015
0.006
1.1110
0
Animal 8* dis-
iodine.
0.3695
0.007
solved in 318 cc.
,
of Ringer's
solution.
0.827 mg. of KI
6
KI
0.4581
0.009
dissolved in
0.4437
0.010
318 cc. of Ring-
er's solution.
* See Table I.
injected and the remaining half of one lobe taken out. Much
more of the iodine from the colloid iodine of these animals was
taken up than from the colloid iodine of normal animals (Table II).
H. B. van Dyke 17
Even here considerable iodine as KI was still bound by the glands
which had taken up some colloid iodine. These results suggest
that the colloid iodine, bound but incompletely synthesized into
active principle, is in a more diffusible form in the acutely iodized
gland than in the more normal resting gland. Moreover, the
incompletely synthesized active principle may be more readily
split by hepatic action (7) into simpler products (e.g., iodides)
which are then bound by the hyperplastic thyroid gland.
Marine and Rogoff (3) declare that even 30 hours after intra-
venous administration of a solution of KI very little of the thyroid's
active principle, as measured by the effects of thyroid on tadpole
growth and metamorphosis, has been elaborated. The less
diffusible and less readily split normal colloid iodine compound
(Table II) probably represents the more fully elaborated active
principle.
SUMMARY.
The findings of Marine and Feiss and Marine and Rogoff that
the hyperplastic thyroid gland of the dog rapidly bincls iodine
intravenously introduced as a solution of KI were confirmed.
By a method already described (5) the ratio value of iodine in
cells to iodine in whole gland was determined and found to be very
low after the intravenous injection of KI solution into dogs with
hyperplastic glands when those glands were removed 1.5 to 60
minutes after the injection. The ratio value more nearly ap-
proached the normal if the interval elapsing between injection
and removal of gland was made about 24 hours instead of 1 hour
or less as in most of the experiments. This finding is in keeping
with the histological changes described by Marine and Rogoff in
such glands 20 hours after the intravenous injection of KI solution.
When iodine as colloid iodine solution of normal animals was
administered intravenously practically none of the colloid iodine
was taken up by hyperplastic glands during the periods of time
used in these experiments; yet from an injection of a comparable
amount of iodine in the form of KI the ready binding of iodine by
similarly hyperplastic glands was proved. Colloid iodine of
hyperplastic glan'ds removed 1 hour after the intravenous injection
of KI solution was taken up to some extent by hyperplastic glands;
but these last named glands bound additional iodine as KI intro-
18 Iodine in the Thyroid Gland
duced after the colloid iodine injection. The incompletely
synthesized active principle is probably more diffusible and more
readily split into simpler products than active principle fully
synthesized.
BIBLIOGRAPHY.
1. Marine, D., and Feiss, H. O., /. Pharmacol. and Exp. Therap., 1915,.
vii, 557.
2. Marine, D., and Rogoff, J. M., J. Pharmacol. and Exp. Therap. , 1916,
viii, 439.
3. Marine, D., and Rogoff, J. M., /. Pharmacol. and Exp. Therap., 1916-17,
ix, 1.
4. van Dyke, H. B., J. Biol. Chem., 1920-21, xlv, 325.
5. Tatum, A. L., J. Biol. Chem., 1920, xlii, 47.
6. Kendall, E. C., J. Biol. Chem., 1914, xix, 251.
7. Blum, F., and Griitzner, R., Z. physiol. Chem., 1920, ex, 277.
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AUGll 1941
rtrT i f 4OvM
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/FCB i;? 1974
v
LD 21-100m-8,'34
52JM2J
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