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Full text of "The Collecting net"

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Vol. XVI, No. 1 



SATURDAY, JUNE 28, 1941 



THE SPRING CRUISES OF THE 
ATLANTIS TO GEORGES BANK 

Dean F. Bl'mpus 
ll'oods Hole OccanogvapJiic Institution 

The .Atlantis made four cruises to Georges 
Uank this spring, one in March, one in April, and 
two in May, in order to continue the studies of 
the productivity of Georges 
Bank, that very productive 
shoal water region extending 
180 miles to the east of Cape 
Cod. This series of cruises 
during the spring months when 
productive rate of plants and 
animals is very great is a con- 
tinuation of a program com- 
menced in the spring of 1940 
under the direction of Dr. 
George L. Clarke, of Harvard 
University, aided by Dr. Gor- 
don Riley of the Bingham 
Oceanographic Foundation of 
Yale University, Dr. Mary 
Sears of Wellesle}' College, 
Mr. George C. Whiteley, Jr., 
of the Hill School, and Mr. 
Dean F. Bumpus of the Woods 

Hole Oceanographic Institu- 

tion. 

As in 1940 a network of about 34 stations was 
made during each cruise over the bank in order 
to measure the tem- (Continued on page 9) 



M' 1^' % €akniet 



The first weekly seminar of the 
season will be held on Tuesday, 
July 8. 



THE PHYSIOLOGY COURSE AT THE 
M. B. L. IN 1941 

Report of the Staff. 
Pliysiology Course 

The Physiology Course is centering its work 
around topics in cellular and comparative physiol- 
ogy of marine organisms. Two two-week periods 
of laboratory are planned. The 
first of these is already pro- 
ceeding under the direction of 
Drs. K. C. Fisher, C. L. Pros- 
ser and F. J. M. Sichel. They 
have designed their individual 
or group student research 
along the lines of Cellular 
Respiration, Comparative 
Physiology of Nervous Sys- 
tems and the Physiology of 
Muscle and Peripheral Nerve, 
respectively. 

- The second two-week peri- 
od of the course will be de- 
voted to studies in Cytochem- 
istry ; Water Balance in Ma- 
rine Organisms ; Properties of 
Cell Memljranes and Blood 
Gas Studies. This will be 
conducted Ijy the new mem- 
bers of the staff, Drs. R. Bal- 
lentine, R. T. Kempton and A. K. Parpart. 

In the above program the wealth of marine 
material available is being exploited to its fullest 



THURSDAY, July 3, 1941 

8:00 P. M. 
M. B. L. Auditorium 

Lecture : 

'Current approaches to the Plant 

Hormone Problem." 

Dr. George S. Avery, Jr., 

Professor of Botany, 

Connecticut College for Women, 

New London, Connecticut 



TABLE OF 

The Physiology Course at the M. B. L. in 1941 1 

The Spring Cruises of the "Atlantis" to 
Georges Bank, Dean F. Bumpus 1 

Relation of Macronuclear Regeneration in 
Paramecium aurelia to Macronuclear Struc- 
ture, Amitosis and Genetic Determination, 
Dr. T. M. Sonneborn 3 

Notes on Eulima oleacea Embryology, George 
M. Gray 4 

National Education Association at Woods Hole 7 



CONTENTS 

Hans Driesch, Philosopher, Ralph C. Busser.... 8 
M. B. L. Department of Chemical Supplies and 

Scientific Apparatus 8 

Physiology Class Notes 10 

Botany Class Notes 11 

Embryology Class Notes 11 

Introducing Dr. Olga Janowitz 12 

Items of Interest 13 

Cold Spring Harbor Symposium 14 

Directory for 1941 15 



June 28, 1941 ] 



THE COLLECTING NET 



extent. It is hoped that by a well-rounded lab- 
oratory plan the interest and enthusiasm of the 
student will be kept at a high pitch. The labora- 
tory work is supplemented by daily lectures by 
the staff and as many as possible of the research 



specialists working at the laboratory. 

Following the first four weeks, the final period 
of the course will entail individual work by the 
students on a problem of his or her choice under 
the guidance of a member of the staff. 



RELATION OF MACRONUCLEAR REGENERATION IN PARAMECIUM AURELIA 
TO MACRONUCLEAR STRUCTURE, AMITOSIS AND GENETIC DETERMINATION 

Dr. T. M. Sonneborn 
Department of Zoology, Indiana University 



In recent years the study of unusual types of 
nuclei has been remarkably fruitful in the field of 
cytogenetics. The present paper is the first step 
in a study of a long known but still almost com- 
pletely enigmatic type of nucleus. 

The nuclei in Parauicciuni and other ciliate 
Protozoa appear in two forms, as micronuclei and 
macronuclei. Every normal individual has at 
least one nucleus of each of these two kinds. The 
micronuclei are in all important respects typical 
nuclei with chromosomes that behave as do chro- 
mosomes in higher organisms. But the macro- 
nucleus, though it develops from a micronucleus, 
shows no chromosomes and no indication of the 
precise mitotic and meiotic nuclear behavior typi- 
cal of other nuclei at the time of cell division and 
gamete formation. Moreover, at times of fertili- 
zation it unravels into a complex skein which 
breaks up into many pieces ; and the latter are 
absorbed in the cell. The lost macronuclei are 
then replaced by new ones formed from products 
of the micronucleus. In spite of its peculiar or- 
ganization and behavior, the macronucleus is 
clearly essential for the life of the cell, while the 
micronucleus, though a typical nucleus, can be 
dispensed with. Of the many perplexing ques- 
tions raised by such a strange situation, the pres- 
ent paper deals primarily with the following : 
( 1 ) How does it happen that the macronucleus 
appears to divide directly without going through 
the complex processes of mitosis? (2) How do 
the micronuclei and macronuclei share and inter- 
act in the determination of hereditary characters? 
As will appear, the answers to these questions, in 
so far as they can now lie given, depend upon the 
discovery here reported of a new and unique 
method of nuclear reorganization. 

Paraniecium aurelia undergoes fertilization dur- 
ing the processes of conjugation and autogamy. 
For present purposes the only important aspect of 
these processes is the behavior of the macronu- 
cleus already mentioned : its unravelling into 



skeins, fragmentation of the skein into pieces, 
absorption of the pieces, and development of a 
new macronucleus from the micronucleus. I have 
found that, at times of such reorganizations, oc- 
casionally the micronuclei fail to produce new 
macronuclei. This of course is inevitable when 
micronuclei are absent, as they sometimes are ; 
Init it also happens sometimes even when micro- 
nuclei are present. In the latter case, there is 
some evidence to indicate that the micronuclei did 
not fuse in fertilization, but remained and multi- 
plied in the reduced condition. In either case, the 
cell commonly solves the problem of getting a 
new macronucleus, so essential for its life, by an 
alteration in the behavior of the many fragments 
of the old macronucleus. Instead of becoming 
al)sorbed in the cell, they grow up into new ma- 
cronuclei. Thus, as many as 35 or more new 
macronuclei may arise from the 35 or more frag- 
ments of the old disintegrated macronucleus. 
These are segregated out to the various cells 
formed at successive fissions, until only one re- 
mains per cell and they then begin to divide at 
fissions like regular macronuclei. This behavior 
is repeated at subsequent reorganizations : the re- 
generated macronucleus or its descendant unravels 
into a skein and breaks up into fragments which 
again regenerate new macronuclei. By inducing 
another reorganization early in the process of 
macronuclear regeneration, before the fragments 
have reached full growth and, indeed, l^efore their 
number has been reduced to one per cell, one can 
observe that the disintegration of these incom- 
pletely developed macronuclei varies with their 
degree of development : when quite small, they 
form no skein at all ; when larger, they form a 
very simple skein ; when still larger the skein be- 
comes more and more complex until at full 
growth it closely resembles the skein of normal 
macronuclei. For comparison, one may observe 
the behavior of still unabsoi"1)ed macronuclear 
fragments in individuals that have in addition a 



The Collecting Net was entered as second-class matter July 11, 193.5, at the Post Office at Woods Hole, Mass., 
under the Act of March 3, 1879, and was re-entered on July 23, 1938. It is devoted to the scientific work at 
marine biological laboratories. It is published weekly for ten weeks between July 1 and September 15 from Woods 
Hole, and is printed at The Darwin Press, New Bedford, Mass. Its editorial offices are situated in Woods Hole, 
Mass. Single copies, 30c by mail; subscription, $2.00. 



THE COLLECTING NET 



[ Vol. XVI, No. 138 



macronucleus formed in the normal way. When 
such animals are induced to conjugate, the old 
fragments never form skeins, but behave just as 
do the regenerating fragments before much 
growth has occurred. These observations dem- 
onstrate therefore, that the complexity of the ma- 
cronuclear disintegration skein reflects a corre- 
sponding complexity in the organization of the 
macronucleus; and, what is more important, they 
prove that the macronucleus is a compound 
structure with each of its component parts (cor- 
responding to the more than 35 fragments it 
forms) containing all that is required for the de- 
velopment of a complete and functional macro- 
nucleus, that is, a complete genome. 

This is the clue to the answer of our first ques- 
tion. The macronucleus appears to divide ami- 
totically because it is a compound nucleus, not a 
simple one; it contains more than 35 sub-nuclei 
and at division all that is required to yield ge- 
netically equivalent functional macronuclei is to 
segregate these multiple subnuclei into two 
groups. Of course multi]>lication of the subnu- 
clear components must take place at some stage 
of the nuclear cycle and this must now be sought 
for, not in the behavior of the nucleus as a whole, 
but in the behavior of the parts within it. Per- 
haps it occurs during the profound and little un- 
derstood changes known to precede the gross 
amitotic division. Further study should now he 
directed on this point. 

Animals in which macronuclear regeneration 
has taken place often die or produce some non- 
viable progeny ; but many live and produce a line 
of descent in which all sulisequent reorganizations 
are invariably by the same method. Such cul- 
tures are always of the same mating type as the 
original parent cell, thus further supporting the 
view I have previously maintained, that the ma- 
cronucleus directly determines mating type. On 
the other hand, a whole complex of new charac- 
ters invariably appears in these cultures : reduced 



size, fission rate and viability ; very short inter- 
reorganization periods ; and extremely intense sex 
reactivity. The direct cause of these characters 
is not fully known, but it is of interest that the 
size and viability appear to be reduced more 
when micronuclei are lacking than when they are 
present, thus implying a direct effect of the mi- 
cronuclei on these characters. Further, the ma- 
cronuclei in such animals are also reduced in size, 
suggesting a direct correlation between macronu- 
clear size and cell size, and an inability of the 
single macronuclear fragment to grow into a 
macronucleus of full size. The latter fact ma}' 
also play a part in determining the low vigor of 
such animals, their strong mating reactivity and 
their short interreorganizational periods. Regen- 
erated macronuclei, though adequate to support 
life and reproduction, may still lack something re- 
quired for perfectly normal functioning. 

The results here set forth show (1) that the 
macronucleus is compound, (2) that the skein 
into which it disintegrates is a reflection of the 
complexity of its organization, (3) that macro- 
nuclei regenerated from fragments are nearly, but 
not quite, perfect and so determine corresponding 
imperfections in the cell, and (4) that inicronu- 
clei, though not indispensible, nevertheless have 
certain direct effects on cell characters. These 
are first steps towards a solution of problems of 
cytogenetics in Parauiccium anrclia ; further steps 
in the same direction should follow from the op- 
portunities here provided for studying separately 
and in various combinations the physiological and 
genetic roles played by micronucleus, normal ma- 
cronuclei and imperfect regenerated macronuclei, 
and from the methods here reported for attacking 
problems of macronuclear organization. 

(Summary prepared for the press of a paper pre- 
sented before the American Society of Zoologists at 
the meeting of the American Association for the 
Advancement of Science on January 1, 1941.) 



INFORMAL NOTES ON EULIMA EMBRYOLOGY BY THE EMERITUS CURATOR 
OF THE M. B. L. MUSEUM 

George M. Gray 



[Note: Mr. Gray has been associated with the 
Marine Biological Laboratory for fifty years as 
collector, curator of the Supply Department and 
lastly as Museum curator.] 



In August 1932 the author published in The 
Collect Tng Net an article on the habits and 
habitat of this dainty little mollusc. There was 
nothing whatever in the article about its breeding 
however. The following embryology comprises 
just a few notes taken during the last two years. 

Eulima olcacca, of Kurtz and Stimpson, or 



Mclanella olcacca of other authors, is a beautiful 
small gastropod mollusc of a creamy-white color, 
with jet black eyes. The shell is finely polished. 
It tapers to a fine point, and the largest one I 
have is barely 10 mm. long or 6/16 of an inch, , 
about 3 mm., 1/8 of an inch, at broadest part 
with 12 whorls, finely and delicately separated by 
lines. It is certainly a dude among our small 
molluscs. It can sometimes be taken by dredging 
but as it is a "quasi parasite or commensal", on 
the black sea cucumber (Thyojic hriarcus), it is 
easier to get them by collecting the sea cucum- 



June 28. 1941 ] 



THE COLLECTING NET 



bers, though one time I handled something like 
250 Thyone and got seven Eiiliiiia, and two of 
these were on one sea cucumber. I have had 
better luck at other times. 

Recent Observations 

July 14. 1939. On the morning of July 8th 
just before noon, one of our collectors brought 
me among other small molluscs, ten Eul'una olea- 
cea, alive and some of them attached to Thyone 
briareus. I took the snails and put them in a 
separate finger-bowl of sea water and let them 
stand. 

The next morning there were in the bowl sev- 
eral small white specks or spots, somewhat small- 
er than the head of an ordinary pin. They did 
not seem to mean much, but on investigation 
proved to be the egg capsules of the Euluna. 
There were evidently a number of individuals 
packed closely together and surrounded by a sort 
of jelly-like substance — something like that which 
surrounds the eggs of the frog. There were only 
a few of these capsules at first. 

On my next looking, which was the following 
morning, I found the number had increased to 
perhaps 18 to 20, w'hich means that a single snail 
laid more than one capsule. About this time I 
took six of the snails and placed them in a sep- 
arate finger-bowl of sea water, leaving four snails 
in the bowl with the eggs. I examined the eggs 
for two or three days. I observed no sign of life, 
but each individual was evidently larger and be- 
ginning to separate from the others. Though no 
movement could be observed with the microscope 
I was using, the eggs were undoubtedly in the 
cleavage stages. (My microscope was not high 
powered enough to show.) On Friday morning 
on looking at them, I found that each individual 
had separated from its companions and all were 
whirling around or moving about more or less 
rapidly. They looked as though they thought 
they were going places, but all this action was 
taking place inside of the svirrounding jelly. These 
individuals were rather perplexing to count as 
they did not stay put. We judged that each cap- 
sule contained from fifty to sixty separate indi- 
viduals and were in what is known as the veliger, 
or larval, stage. 

The eggs were laid on the bottom and sides of 
the finger-bowl below the surface of the water. 
Up to date (July 14) have noticed no eggs from 
the six snails which I had separated from the ori- 
ginal lot. I doubt if any more eggs have been 
laid by any of the ten original snails. Friday af- 
ternoon of the 14th I placed a Thyone in the bowl 
which had the six Eulima. In about a half hour 
or perhaps longer, two of the snails had settled 
on the Thyone, and in less than four hours, all 
but one were on the same sea cucumber. The last 
three to go on may have gone on in less than the 



four hours as I was busy at other work, and did 
not keep too close a watch. 

July 15. The next morning all six were on the 
Thyone. I then took the Thyone out, removing 
the snails, and put all the snails in a bowl of clean 
sea water with no Thyone to see if by chance 
their night's repast had put them a better humor 
for laying or if they had laid out their supply. 

I washed the Thyone to see if there were any 
eggs laid on it, but oljserved none in the wash 
water. They would be difficult to find on the 
Thyone. In the meantime there seemed to be 
two clusters of fresh laid eggs. The earlier eggs 
were in the veliger stage, while those that seemed 
fresh laid were very quiescent, proliably in the 
cleavage stage. ( These two capsules of fresh eggs 
were in the bowl with the original eggs and not 
in the Thyone lot.) 

The eggs which I first found in the veliger 
stage were seemingly more energetic than ever 
the morning of the 15th, still in the jelly but the 
whole capsule, or capsules were enlarged over 
what they were at first (or larger than they were 
on the 14th). The individual veligers seemed 
larger also. It would seem that the walls of sur- 
rounding jelly had been inflated or pushed out all 
around, something like a toy balloon is made 
larger. This of course would create a greater 
space for the veligers to perform their gyrations, 
though they still liumped elbows. 

July 16 — past 9 A. M. Veligers larger, still 
in the capsule, and still think they are going 
places very rapidly. This morning I discovered 
six egg capsules in the bowl with the six Eulima 
which I had taken away from the Thyone yester- 
day. I think one was laid yesterday before 6 
P. M. These were evidently in early cleavage 
stages. There was no individual movement. 
These capsules were on the sides of the bowl. At 
8:30 P. M. there were ten egg capsules in the 
finger-bowl, three of them on the bottom of the 
bowl. None of the first laying had left the jelly 
but had increased in size especially as individuals. 
Very active. On examination none of the fresh, 
or last laid eggs, had shown any separate individ- 
ual motion or swimming, but were all adhering 
together. 

July 17 — 8:30-9:30 A. M. None of the veli- 
gers have left the mother capsule as yet. Still 
very lively and seemingly larger. Does not seem 
as though they could get along in their cramped 
home much longer without serious friction. In 
the finger-bowl containing the six Eulima which 
had been on the Thyone there were twenty-one 
egg capsules as against the ten of last night. Of 
the eleven additional, most of them were on the 
sides of the bowl, one or two barely covered with 
water, one at the very surface. Some of the last 
laid were very fresh, evidently laid early this 
morning. All the capsules were still holding the 



THE COLLECTING NET 



[ Vol. XVL No. 138 



individuals closely packed. No veligers yet 
(10:15 A. M.). 

At 6 P. M. one more capsule of eggs with the 
six Euliuia still closely packed, but evidently be- 
ginning to separate more, but none free-swim- 
ming. No veligers, though one had clearly sep- 
arated from the others in one of the capsules. 

July 18 — 9 A. M. There were twenty-eight 
capsules of eggs in the finger-bowl with the above 
six Euliiiia. Beginning to show clearer individual 
outlines. Still growing, but not veligers. The 
Thyove feed was evidently an inspiration to the 
Eulhna. 

Twelve fresh Eiiliiiia were collectly yesterdav 
on Thyonc. separated from the Tliyonc and placed 
in a finger-bowl of clean sea water, liut there were 
no eggs this A. M. None of the earliest laid eggs 
have left the home brooder as yet. Still in the 
veliger stage and going strong. 

At 5 ;45 P. M. evidently not much change in 
the above eggs. No veligers have come forth 
from the capsules. The snails collected yesterday 
have not laid. Have been partially changing water 
daily on all eggs. 

July 19 — S A. M. \'eligers in first lot of eggs 
.still going strong l)ut still in the doghouse. Seems 
to be no indication of coming out. A few more 
&gg capsules were in the bowl with the same six 
EuHiiia as mentioned yesterday. As I disposed of 
a half dozen yesterday, it would mean that about 
ten were laid last night and late P. M. yesterday. 
No eggs as yet from the Eulima collected on the 
17th. This morning I put the four Eiiliiua 
(which had been a part of the original ten) into 
a finger-bowl of fresh sea water, into which I had 
first placed a small Thyonc. These four had been 
in the same bowl since first collected and in the 
last two or three days had been quite negligent 
about laying. It may Ije that a good feed on the 
Thyonc will pep them up to laying eggs again, 
like some humans who flourish at others' expense. 
It is only about a half hour ago that I put them 
in with the Thyonc and three have already at- 
tached to the Thyonc. At 5 :45 still three on the 
Thyonc, one off. May have been on and gone off 
again, but think not. Of the twelve collected the 
17th I gave away two so that I have ten left to 
care for. At 5 :45 P. M. one egg cluster or cap- 
sule was found with these ten. At 5 :45 P. M. 
oldest ones still in veliger stage and inside the 
original ca])sule. Of the capsules from the six 
Eulima, the individual eggs have separated more, 
but none free-swimming, i.e., no veligers ; perhaps 
two or three additional capsules. 

July 20 — 9:30-10 A. M. Evidently the veli- 
ger stage with some of the egg capsules mentioned 
last took place last night or early this A. M. as 
some individuals of these eggs were freely swim- 
ming, while others were just barely showing per- 



ceptible movement. (These are from the six 
Eulima which had been separated originally.) 
The Eulima themselves had possibly not laid any 
more eggs since last night. Of the first lot of 
veligers some seem dead while others are very 
slow in their movements. The ten snails collected 
the 17th had laid six capsules of eggs as I found 
this A. M. I took these ten Eulima, put them 
and a small Thyonc into another finger-bowl of 
sea water and now await developments (10:15 
A. M.). 

About noon today noticed the first coming out, 
or beginning to come out, of the veligers from the 
capsule, only a few. At 5 :40 P. M. there were 
some with quite lively veligers. Some just mov- 
ing and .some not, the livelier ones are older. 
These were from the six Eulima which had been 
separated from the first ten of the 8th, but no 
Eulima on the Thyonc of the ten put in this 
A. M. (This would mean that twelve days at 
least from egg to coming out of the capsule.) One 
went on but is off tonight. 

July 21 — 9 A. M. This morning I found the 
wall on one side of one of the oldest egg capsules 
was open and a number of veligers or well-ad- 
vanced larvae were swimming around freely in 
the water. I think the veligers had pushed out 
this wall and were taking advantage of their es- 
cape from the old homestead and enjoying a 
swim, though not in forbidden waters. Four out 
of the ten Eulima were on the Thyonc, the other 
six were sulking on the sides of the finger-bowl, 
fi\-e were under water and one barely above the 
surface. 

July 22 — S:30 A. M. This A. M. four of the 
ten last collected Eulima were on the Thyonc. 
The other six were mostly on the sides of the 
bowl. But no eggs were found either on the 
Thyonc or on the sides of the bowl. Water was 
changed and the animals left as before, and of the 
four Eulima of the first lot, no eggs were found. 
The Thyonc had eviscerated. I threw out this 
Thyonc. washed the inside of bowl, put in another 
Thyonc with the snails, added fresh sea water and 
left them to their fate. To lay or not to lay — 
that is the burning question. 

No more coming out parties, no more debu- 
tantes, no more veligers coming through the ma- 
ternal wall of the egg capsule. Those which were 
in the veliger stage were going lively. These are 
the ones from the six Eulima which were of the 
first lot collected on July 8th. Those that left the 
capsules yesterday are about as they were last 
night. 

July 23 — 9:30 A. M. Found no fresh eggs 
from any of the Eulima. The two lots on the 
Thyonc in different bowls had no egg capsules, 
neither on the Thyonc nor on the bowl. These 
lots were : one lot of four from the catch of July 



[line 28, 1941 ] 



THE COLLECTING NET 



8th, one lot of ten EuVnna from the collecting of 
July, 17th. The veligers from the lot of six Eii- 
liiiia are still lively and some have come out of 
the capsule and are swimming ecstatically about 
in the water outside though most of them are in 
the capsule. 

Many of the veligers were out skylarking 
around in the water outside of the capsules, seem- 
ingly having a grand outing. Several, three at 
least, of the capsules were empty with one excep- 
tion, which had only a few. The others will have 
to be examined under a better light than tonight, 
though I did not notice any new eggs. 

July 24. Things seemed to be about as yester- 
day. Some veligers of the first lot appeared dead. 
Of the egg capsules in the bowl containing the six 
Euliiua of the original lot they seem to be in about 
the same condition as yesterday, but of course a 
little more advanced. Some in, some out of the 
capsules. Discovered no eggs in the lot of four 
Euliiua with Thyone, or in the lot of ten. Added 
fresh sea water and left them. 

July 25. Noticed no particular change in any 
lot and found no fresh eggs in any lot changed as 
above. No eggs for several days. 

July 28. The ten Eidima collected the 17th and 
which had been on the Thyone and then taken ofif 
and put in a finger-bowl by themselves lately had 
two egg capsules in with them. I discovered this 



A. M. some of the other veligers are still living. 

August 4. So far as I could judge no new eggs 
of Euliiiia have been laid for several days. On 
July 31st six Euliiua just collected were brought 
to me and put in a separate dish. On August 1st 
and 2nd more just collected Euliiua were brought 
to me and on August 2nd one more freshly col- 
lected Euliiiia was given to me. These were all 
placed in the same di.sh with those of July 31st. 
and could not discover that any of them had laid. 

On August 3rd two additional freshly collected 
Euliiua were left in a three-ounce corked vial of 
sea water overnight ; I found one or two egg cap- 
sules on the side of the vial. I at first thought 
only one but on pipetting them out was surprised 
to find two whole capsules. These two Eulima 
and their eggs I am keeping in a separate finger- 
Ijowl from the others. 

August 6. Two tgg capsules in the bowl of 
six Euliiua. None from last collected Eulima. 

August 11. I think it was on August 7th or 
not later than the 8th that I gave the last collected 
lot of Eulima (those of July 31st and August 1st 
and 2nd) a change to a bowl of sea water con- 
taining a Thyone. Up to this time none of this 
lot had laid eggs. There were very small patches 
or spots of jelly or mucus, but no well authenti- 
cated eggs in them. 

(Concluded Next IVeck) 



THE NATIONAL EDUCATION ASSOCIATION AT WOODS HOLE 



At the suggestion of Mabel Studebaker, for- 
merly a worker at the Marine Biological Labora- 
tory, and now Northeastern Regional Director of 
the Classroom Department of the National Edu- 
cation Association, the executive board of that 
organization decided to hold its business sessions 
at Woods Hole for the two days preceding the 
Boston Convention. 

Dr. Charles Packard of the Marine Biological 
Laboratory very generously extended the facilities 
of his institution to the group. Meetings were 
held in the Committee Room of the Administra- 
tion Building. The ten members of the commit- 
tee representing teachers from the four corners of 
the United States were housed in the new dor- 
mitory. 

On Thursday afternoon, when the last member 
had arrived, the Executive Board seated around 
the table were : 
Mrs. Mary D. Barnes, President of the Department, 

from Elizabeth, New Jersey. 
Miss Margery Alexander, Vice-President, Charlotte, 

N. C. 
Mrs. Eleanor F. Edmiston, Secretary, San Diego, 

Calif. 
Miss Mabel Studebaker, Northeastern Regional Di- 
rector, Erie, Pa. 



Miss Katy V. Anthony, Southeastern Regional Di- 
rector, Richmond, Virginia. 
Harold H. Blanchard, North Central Regional Direc- 
tor, South Bend, Ind. 
Miss Florence B. Reynolds, South Central Regional 

Director, Omaha, Neb. 
Miss Mary E. Bond, Northwestern Regional Direc- 
tor, Bellingham, Wash. 
Wilber W. Raisner, Southwestern Regional Director, 

San Francisco, Calif. 
Miss Elphe K. Smith, former President and Director 
ex officio, Tigard, Oregon. 

The Department of Classroom Teachers is the 
largest of the twenty-seven departments of the 
National Education Association. It is composed 
solely of class room teachers. They comprise 
80% of the National Education Association's total 
membership, which includes over 200,000 educa- 
tors. All teachers, public and private, in the 
United States or in its possessions are eligible to 
membership in the Association and to consequent 
membership in the Department of Classroom 
Teachers. 

The Department works constantly for the ad- 
vancement of the teaching profession. Its service 
to the teachers of this country in the establish- 
ment of adequate salary and retirement benefits 
has been inestimable. — Mary D. Barnes. 



THE COLLECTING NET 



[ Vol. XVL No. 138 



HANS DRIESCH, PHILOSOPHER AND SCIENTIST 

Ralph C. Busser 

American Consul General at Leipsig, Germany, until 1940, 
Philadelphia, Pa. 



On the 16th of April, 1941, Hans Driesch, the 
world renowned German philosopher and scien- 
tist, died in his 74th year at Leipzig, Germany. In 
Octoljer, 1933. he had retired from his position 
as professor ordinarius at the University of Leip- 
zig, receiving from the Saxon Ministry of Edu- 
cation the title of "professor emeritus." Driesch 
stands out as one of the greatest thinkers and 
educators of the world, and very few university 
professors have done as much in promoting in- 
tellectual cooperation lietween nations and a sym- 
pathetic understanding of the mentality, ideals and 
achievements of his own and other countries in 
the realm of science and culture. 

Hans Driesch was born in 1867 at Kreuznach 
(Rhineland). His father was a wholesale mer- 
chant in Hamburg, where he got his early edu- 
cation. Hans Driesch, who became a doctor of 
law, medicine and philosophy, began his career 
as a zoologist and was engaged in research work 
at the zoological station in Naples from 1891 to 
1900. Later he devoted his studies to philosophy, 
teaching it from the viewpoint of natural science 
rather than as a dogmatic metaphysician. His 
most famous work is "The Science and Philoso- 
l)hy of Organism" (1907/8). which was first 
published in Great Britain, as it consisted of the 
"Gififord Lectures" for which he received the 
(jitford Prize at the University of Aberdeen, 
where he taught philosophy from 1907 to 1908. 

In 1909 Driesch became a Privatdozent (lec- 
turer) at the University of Heidelberg, where in 
1911 he was appointed jirofessor extraorclinarius. 
In 1912 he ]3ublished his second principal work 
called "Ornungslehre" (Logic), then in 1917 the 
"Wirklichkeitslehre" (Metaphysics), both of these 
works together forming a complete system of 
philosophy. In 1920 Driesch became professor 
ordinarius of philosophy at the University of 
Cologne. 

In 1921 Driesch was appointed to the same 
])osition at the University of Leipzig, but in 1923 
he was abroad for more than a year giving lec- 
tures in China, Japan and the United States. 
While in China, Driesch was guest professor at 
the Chinese State University in Nanking and 
I'eking, where he succeeded Professor John 
Dewey, the famous American writer on philo- 
sophic and political subjects. 



In recent years Professor Driesch has taken a 
great interest in the problem of psychology and 
wrote "Leib und Seele" (Mind and Body), and 
"The Crisis in Psychology" (Princeton, 1924). 
Driesch is also famous for his studies in psychical 
research. His book on "Para-psychologie" cre- 
ated a sensation in the scientific world. 

In 1926 Driesch was granted a term's leave of 
alxscence from the University of Leipzig, which 
period he spent at the University of Wisconsin in 
Madison, to which he was appointed "Carl 
Schurz Memorial Professor". He has also lec- 
tured at the English universities of London, Man- 
chester, Leeds and Cambridge, at the national 
LTniversity in Buenos Ayres, in Italy, Switzer- 
land, Czecho-Slovakia, and other countries. 

Until his recent retirement Professor Driesch 
was Director of the Philosophical Institute at the 
University of Leipzig. He received the title of 
honorary doctor at a number of universities, was 
formerly president of the Society for Psychical 
Research in London, and up to the time of his 
death was a member of many other scientific and 
learned societies. Driesch was a fine linguist, 
speaking English, French and Italian fluently as 
well as his own mother tongue. 

Driesch was one of the most popular professors 
at the University of Leipzig, students flocking 
there from all parts of the world to listen to his 
lectures. He was an international personage not 
only through his experience as a lecturer at num- 
erous American and foreign Universities, but 
more through his celel)rated works on philosophy 
and psychology, which have challenged old theo- 
ries and opened up new paths of thought. 

During the past ten years I had many conver- 
sations with Dr. Driesch in Leipzig, where I was 
stationed as American Consul General until my 
retirement from the Foreign Service in January, 
1940. As Dr. Driesch never accepted the Nazi 
philosophy but clung to his lifelong belief in lib- 
eral and democratic principles, he was never per- 
mitted, after 1933, to lecture or hold public ad- 
dresses in his country. The loss of academic 
freedom in Nazi Germany, even in non-political 
fields of learning, and other restrictions upon in- 
tellectual and religious activities had a stifling 
effect upon Dr. Driesch, whose literary work be- 



June 28, 1941 ] 



THE COLLECTING NET 



came practically sterile. No philosophy except 
that of despair could thrive in such a prison-like 
atmosphere. 

Meeting Dr. Driesch frequently at our respec- 
tive homes and elsewhere in Leipzig, I greatly 
enjoyed his friendship and learned to appreciate 
his splendid character, broad-minded views, and 
intellectual and social qualities. Dr. Driesch was 
a popular figure in University and other circles 
in Leipzig, and his conversation was always 
sprightly and entertaining. Although one of the 
greatest intellectuals of his time, he never made 



a display ef his knowledge in society, rather talk- 
ing on subjects most interesting to his par- 
ticular listener. Dr. and Mrs. Driesch frequently 
entertained at their home, which was on the floor 
above my home in Leipzig, and their guests were 
always sure of a hearty welcome and the tradi- 
tional German hospitality. Their children, Kurt 
and Ingeborg Driesch, have already achieved con- 
siderable success in the musical world, the former 
as a conductor and composer, and the latter as a 
violinist ; each of them has held many concerts in 
Germanv during recent vears. 



THE SPRING CRUISES OF THE ATLANTIS TO GEORGES BANK 

(Continued from page 1) 



perature and salinity of the water at regular in- 
tervals. From these same levels samples of water 
were taken in order to measure the concentration 
of Oo, Nitrate, Phosphate and chlorophyll as an 
indicator of the abundance of plant pigment which 
in turn is an indicator of the abundance of phy- 
toplankton. Samples of sea water were also taken 
from these same levels for numerical and taxo- 
nomic studies of the phytoplankton and nanno- 
plankton species, the latter in which Dr. James B. 
Lackey, of the U. S. Public Health Service, is 
interested. 

Oblique tows for zoo-plankton, copepods, 
schizopods and decapod larvae, other small crus- 
tacean forms, sagittae. Haddock eggs and larvae, 
were made with the recently developed quantita- 
tive Plankton Samplers from two or three strata 
depending on the depths in order to sample the 
whole water column. Other tows were made from 
the bottom to the surface with meter and a half 



stramin nets in order to sample the larger mem- 
bers of the plankton population. 

It is hoped that not only will a good picture of 
the productivity of the region be develo])ed, but 
that factors influencing the survival of young 
haddock be obtained. At present it is not at all 
clear whether biological or physical factors or 
Ijoth are responsible for the large loss of young 
haddock in this peculiar marine aquarium. 

We hope soon in the future to continue the 
collection not only of the plankton on Georges 
but also a study of the fauna living on and in the 
first few centimeters of the bottom. Photographs 
made last spring by Dr. Maurice Ewing of the 
Ijottom of Georges Bank have whetted our inter- 
est and enthusiasm for a complete survey of this 
phase of the problem which is so important in the 
economy of the sea. We plan this summer to 
develop a clam shell dredge for just such a quan- 
titative study. 



DEPARTMENT OF CHEMICAL SUPPLIES AND SCIENTIFIC APPARATUS 



Apparatus, Room 3; Chemicals, Room 8 
Office, Room 1, Marine Biological Laboratory 

This department loans without charge to prop- 
erly qualified research workers at the Marine 
Biological Laboratory, chemicals and ordinary 
glassware in reasonable amounts. Certain types 
of special apparatus, required by investigators for 
their research are also available for short periods 
of time, depending on the prior allocation and ad- 
vance requests for such a])])aratus. 

Members of classes are not entitled to supplies 
other than those provided in their regular class 
work. Beginning investigators will receive sup- 
plies only on the authorization of the person un- 
der whom they are working for the season. 

Expensive and fugitive sui^plies such as, liquid 
air, dry ice, and gas mixtures, are paid for by the 
investigator. Chemicals in large quantities and 
those not generally carried in stock, if expensive, 
are also charged. 

In order to carry out the usual services, the co- 



operation of investigators is urged in the three 
following requests : That 

1. Supplies and apparatus no longer in use be 
retvn-ned. In emergencies these may be recalled 
for redistribution. 

2. Glassware, including aquaria and museum 
jars, be cleaned thoroughly before returning to 
the Chemical or the Api^aratus Room. Failure to 
cooperate in this regard means added costs and 
subsequent loss of efficiency and service. 

3. If certain supplies from the Chemical Room 
are to be used by an investigator during the next 
succeeding summer he may reserve them when 
arrangements can be made to do so. Supplies 
thus required must be packed in boxes or cartons 
and properly labeled. A Kept Out Card, obtain- 
able from the Chemical Room for this purpose, 
must be properly filled in and left with these con- 
tainers. All supplies not thus listed, packed, and 
marked, will be returned to stock. 



10 



THE COLLECTING NET 



[ Vol. XVL No. 138 



PHYSIOLOGY CLASS NOTES 



"This is a course in Physiology," quoth Dr. 
Parpart, as we. on the morning of June 17th, sat 
in the lab patiently awaiting our first day of work. 
After our first week of strenuous labor in said 
laboratory we have come to agree not only with 
Dr. Parpart, but also with the janitor who said, 

quote " that course in slopology;" unquote. 

One can easily understand the janitor's point of 
view when, at the end of the day, we observe the 
tidy lalj, with the remains of starfish, frogs and 
dead dogfish hanging out of the waste liaskets, 
and the rest of the day's rubbish ankle deep on 
the floor. It is without a doul)t the janitor's 
paradise. 

The ]5revaihng atmosphere in our lab is, of 
course, one of peace and calm, — with someone 
quietly yelling out manometer readings, the dog- 
fish splashing water over everyone in protest of 
their tank existence, the embryologists looking 
for things we never heard of. Bill Keezer scratch- 
ing his 1)eard, the centrifuge centrrfuging and the 
aspirator aspirating. 

If Mr. Warlnirg were only here he certainly 
would l)e proud of Dr. Fisher's busy students. 
They have run the Warburg apjjaratus almost to 
the point of exhaustion, as observed liy the mighty 
screech it developed and the more than once 
liroken belt. And the results ! — Enough to make 
Mr. Warburg sit up and take notice. To quote 
the cellular respirationists : — "The cells respire 
while we perspire." 

And on the other side of the lalj, trying to con- 
centrate between the ten minute readings of the 



Warburg enthusiasts, we find the quieter type of 
student daintily gaining entrance to a limulus 
with a can-opener, eagerly trying his surgical 
skill on an unsuspecting dogfish, or solemnly 
studying his waves on a smoked drum. These 
are Dr. Prosser's students. 

Back in a little room all by themselves we find 
half of Dr. Sichel's students — all tangled up in 
electrical wirings and muscles. The other half 
are in the air-conditioned dark room, avoiding 
the summer heat, measuring tensions developed 
by frog muscles, and developing a few of their 
summer snapshots on the side. 

This year the laboratory work has been organ- 
ized into seven sections. For the first two weeks, 
three sections have been running parallel, and 
for the last two weeks, there will be four sections. 
A wide choice is therefore given to each student 
as he may select two sections for each two week 
period. 

The stafif of instruction has been changed con- 
siderably this year. With the departure of Drs. 
Hober, Irving, and Shannon, have come Dr. A. 
K. Parpart, Associate Professor of Biology, 
Princeton, to take charge of the course ; Dr. Rob- 
ert Ballentine, Procter Fellow, Princeton ; and 
Dr. R. T. Kempton, Professor of Zoology, Vas- 
sar. 

One thing it didn't take us long to learn is that 
to be a physiologist one must be not only a biolo- 
gist, init also an electrician, a mechanic, a chem- 
ist, a physicist, and, aljove all, an optimist. 

— /. E. H. 



BOTANY CLASS NOTES 



Skepticism, instilled liy certain memliers, is fast 
becoming the outstanding characteristic of this 
year's botany class. Accusations of overacting 
imaginations are many and severe, with the result 
that "Are you sure you saw that?" and such dis- 
paraging remarks are fast becoming botany by- 
words. Though they began quite locally and were 
only sporadic, the skepticism is increasing in di- 
rect proportion to the difficulty of the algae. 

Tuesday. June 17, was the first day of a course 
that wasted no time in going full-steam ahead into 
work. Prefaced in the morning by a short intro- 
duction from Dr. Taylor, not without the inter- 
polation of some of his inimitable quips, the study 
of the "Morphology and Taxonomy of the Algae" 
started with observation of gametes and zoospores 
of Chlamydoinonas feverishly dashing across the 
slide like army men heading for town on a day 
off, and those who had never before met Chlamv- 



doiiioiias can now recognize that primitive algal 
cell form with its typical cup-shaped chloroplast, 
red eye-spot and two flagella. It is fast being 
discovered that a so-called "day's schedule" really 
includes half the night as well, and yet, Bill Gil- 
Ijert, the assistant, promises that "the worst is 
yet to come." 

Field trips, it seems, are the dessert of the 
course, and the first one, held last Friday, was 
all that the algologists had been warned it would 
be. So the class was eagerly anticipating the first 
one via boat to have been held Wednesday, the 
day after this report was due. However, the an- 
ticipation was slightly flavored with fervent pray- 
ers for a sunny day, after the build-up Dr. Taylor 
gave about the lunch question. The Mess, it 
seems, is quite willing to pack a lunch for its pa- 
trons, but it dislikes having excess food on hand 
because of rain, so that once the lunch is made. 



June 28, 1941 ] 



THE COLLECTING NET 



11 



it's made, and the multitude either eats it that da}' 
or holds it for the first decent day thereafter. If 
that day doesn't arrive for several moons or so, a 
stale Mess lunch is just another of those things 
one meets in a good botany course at the M.B.L. 
C'cst la vie, c'est la vie. 

The first field trip, already mentioned, luckily 
fell on a beautiful day, and the initiation was 
spoiled by neither rain nor cold, or even by chilly 
winds. Following the speedy pace set by Dr. 
Runk and Dr. Tseng was quite invigorating and 
seemed second nature to a few people, but it was 
soon evident that most of the class was of the rear 
guard variety. 

The general collecting technique, one soon dis- 
covered, is to wade into the water or muck of the 
])articular habitat, snatch up a few samples, get a 
"squeezing" or two, scribble a label, and then 
dash off on a dog-trot to catch up again with the 
leaders. It was when the first Rhus toxicoden- 
dron was encountered that the preliminaries of 
thoroughly covering one's exposed anatomy with 
the special poison ivy preventative, which gives 
such a lovely yellow jaundice or golden tan effect 
— depending on the individual's aesthetic sense — 
seemed less humorous and more of a pretty good 
insurance. 

Highlight of the trip was the visit to Cedar 
Swamp — a real swamp, by gar, through which 
one carefully kept well within the limits of the 
knee-deep, brown-water trail, or else sank his er- 
rant foot into the depths of nothingness. 



Friday being a special day for the Ijotanists, 
what with the field trip and all, ended up with a 
bang by the first seminar meeting, at which Dr. 
Taylor showed some fine movies — he claims to 
have been an amateur when he took them, but 
that tale is hard to believe — and he kept up a 
running commentary of informative and funny re- 
marks so that several hours were spent as none, in 
looking at algae, tropical flowers in gorgeous 
colors, and details of the Allan Hancock Explora- 
tion Trip ?^2 to Central and South America. 
(General note: If there's a chance for everyone 
to see these films, no one should miss it.) And 
so a wonderful time was had by all. substantially 
augmented afterwards by tea, Ritz cracker sand- 
wiches, and cookies, an elaboration of the regular 
daily 10 p.m. custom which offers time out to late 
lab workers. 

Of all the interesting things viewed through a 
lens to date, it must be admitted that one of the 
best was a submarine suddenly seen by Dr. Runk. 
Indeed, its rare appearance in the waters of 
Woods Hole was sufficient reason to give those 
who were truly scientific-minded an extra field 
trip to Juniper Point for closer observation. 
Rumor has it that a car loaded with ten people — 
only girls visible — caused family difficulties when 
viewed unexpectedly by the owner's wife. But 
it's these unexpected events that give the class 
good subject matter for dinner conversation while 
they're waiting for the tenth refill on the beans. 
— /. W. and C. S. 



EMBRYOLOGY CLASS NOTES 



Dr. Goodrich opened the course in Embryology 
with a bang bright and early on the morning of 
June 17, 1941. After recalling to our so-called 
minds the history and terms with which some of 
us had been vaguely familiar in the past. Dr. Cos- 
tello was introduced and we were plunged into 
the entangling alliances of sperm and egg. The 
Panzer divisions of the sperm being eventually 
victorious we moved on to cleavage and the prob- 
lems of the Germinal Vesicle. Following recent 
work in Amphibia certain talented members of 
our group removed these vesicles very success- 
fully, at first with trepidation and later with 
greater facility. The animal under observation 
was Nereis limhata which we observed with 
mixed emotions in its natural nightly activities at 
the dark of the moon. 

The main part of the week was concerned with 
the highly interesting Teleost fish, species Fun- 
diiliis heteroclitus. The struggle was long and 
drawn out especially when there was a traffic jam 
in the vitelline veins, which only Dr. Goodrich 
could straighten. 



Dr. Schotte ended the first week with a pre- 
cedent-shattering lecture for those brought up 
upon the theory of concrescence and was nobly 
aided by one battered chapeau which in the course 
of the lecture was caused to gastrulate, envagin- 
ate, delaminate, and enfiltrate. The chapeau re- 
covered but the class is still considering. 

Social life which started with a few hesitant 
dips into a cold sea and tennis by George 
and Neil (without a net) received a new lease on 
life at the mixer Saturday night. It really was 
a mixer! "Cutting in," an American custom at 
dances, was strongly objected to by some of our 
more distinguished members because of variances 
from the international norm. 

The class wishes to express its gratitude to its 
two superior assistants — to the fatherly benevo- 
lence of Ray the success of our observations is 
largely due. As for Trink, it has been encourag- 
ing to see such a brilliant mind in all phases of 
rest and play. 

— Patricia Hollistcr and EUzabetli Kirkpatrick 



12 



THE COLLECTING NET 



[ Vol. XVI, No. 138 



The Collecting Net 

A weekly publication devoted to the scientific work 
at marine biological laboratories. 

Edited by Ware Cattell with the assistance of 
Boris I. Gorokhoff and Judy Woodring. 

Entered as second-class matter, July 11, 1935, at 
the U. S. Post Office at Woods Hole, Massachusetts, 
under the Act of March 3, 1879, and re-entered, 
July 23, 1938. 

Jlnti-ohiictng 

Dr. Olga J.\nowitz. formerly Secretary of Uni- 
versity Extension Courses in the University of 
Vienna and Lecturer in Biology in a Realgymna- 
sium in Vienna ; now Instructor of mathematics 
and Latin, Potomac School, Washington, D. C. 

Dr. Janowitz is a native of \'ienna, Austria, 
where she studied at the University of Vienna 
and received her doctorate of jihilosophy in both 
zoology and botany. 

After she received her degree, she taught in a 
Realgymnasium in \'ienna, which was attended 
by girls eleven to nineteen years old. and was also 
appointed by the school department in charge of 
training teachers in biology for Austrian second- 
ary schools. While she was also interested in 
conducting research, she found that her inclina- 
tion was for adult education. She felt that the 
]irinciples of biology would be extremely useful 
in providing a ll'cltaiiscliauiiiu/ for the disillu- 
sioned and bewildered people of post-war Aus- 
tria. In this connection she became secretary of 
the University Extension Courses and there she 
gave a series of lectures each year on biology, 
genetics, and heredity. Prominent in her lectures 
were the arguments that there was no dictator- 
ship in nature and that there was no biological 
basis for racial prejudice. 

Stich lectures naturally lirought her into con- 
flict with the Nazis after they effected the Ansch- 
luss in 1938, and she left Austria a year later for 
England. There, tmder a grant from the British 
Federation of University Women, she condticted 
research with Dr. H. Hardy at the Hull LTniver- 
sity College. He had devised apparatus to col- 
lect jilankton off the shores of England. This 
work had hardly started when it was brought to 
a standstill by the outbreak of war. 

In March of last year she arrived in the United 
States, and in a few weeks she obtained a posi- 
tion in the Potomac School in Washington. She 
is interested not only in teaching, but in various 
other types of work. For instance, this summer, 
in addition to condticting research at Woods Hole, 
she is organizing a group to practice German 
conversation based on biological topics, which is 
meeting weekly in the Old Lecture Hall. Through 
this type of work she hopes to find persons who 
will be able to visit Europe after the war to 
s]3read democratic ideals there. 



M.B.L. CLUB SPONSORS MIXER 

The social season of the M. B. L. Club was 
opened last Saturday night with a mixer, to which 
everyone was invited. Miss Lucille "Sunny" 
Nason, the general chairman, saw to it that the 
long-anticipated event lived up to all expectations. 

Upon entering the club, each person was given 
a name card to wear during the evening. The 
drawings of whales, plant cells, etc., to designate 
the various classes were done by Mary Chamber- 
lin, Roger Cole, Arthur Woodward, and Mrs. T. 
H. Bullock. 

Immediately catching the eye of newcomers 
were the unique decorations of fish nets suspend- 
ed from the ceiling, holding sea urchins, horse- 
shoe crabs, and other sea animals. The Supply 
Department of the Laboratory furnished the ma- 
terials and those who arranged them were Dick 
Lee, Dr. W. \\'. Ballard, Dr. C. Hyman, Warren 
Healy, Dr. Lauren Gilman, Barbs Schraff, and 
Mary Donovan. 

While chatting with variotis groups of young 
and old, the guests were served refreshing punch 
and cookies. Those in charge of the refreshments 
were Mrs. H. B. Goodrich, Mrs. T. H. Bullock, 
Mrs. Edward Warner, H. Duncan Rollason, 
Theodore Genther, Airs. Karl Wilbur, Jane 
Henry, and Jacqueline Waldron. 

Dr. E. R. Brill. Dr. K. M. Wilbur, and Mr. 
Richard Henry had arranged for the latest rec- 
ords to be played on the phonograph and before 
long the gathering turned into an informal dance. 

As a result of the mixer, everyone met a host 
of new people and newcomers began to feel more 
a part of the Laboratory colony. 

Mrs. K. M. Wilbur has been appointed the 
new hostess for the M.B.L. Cltib this year. 



CURRENTS IN THE HOLE 

At the following hours (Daylight Saving 
Time) the current in the Hole turns to run 
from Buzzards Bay to Vineyard Sound: 
Date A. M. P. M. 

Tune 28 7:04 7:20 

June 29 7:50 8:09 

June 30 8:37 9:01 

July 1 9:29 9:56 

Tuly 2 10:23 10:56 

julv 3 11:18 11:59 

July 4 12:19 

Julv 5 12:59 1:16 

Julv 6 1 :56 2:12 

July 7 2:54 3:08 

In each case the current changes approxi- 
mately six hours later and runs from the 
Sound to the Bay. 



June 28, 1941 ] 



THE COLLECTING NET 



13 



Dr. Arthur K. Partart, in charge of the 
Physiology Course of the Marine Biological Lab- 
oratory, has been promoted from assistant to as- 
sociate professor of biology at Princeton Univer- 
sity. 

Dr. Rita Guttman was promoted from tutor 
in physiology to instructor in physiology at 
Brooklyn College on the first of this year. 

Dr. Carl G. Hartman, of the department of 
embryology of the Carnegie Institution of Wash- 
ington at Johns Hopkins University, has been ap- 
pointed professor of zoology and head of the de- 
partments of zoology and physiology at the LTni- 
versity of Illinois beginning September 1. 

Professor Robert Chambers left for New 
York on Thursday. At Washington on Saturday 
he will take part in a conference on shock under 
the auspices of the medical division of the Na- 
tional Research Defense Committee. 

Fourteen persons attended the first meeting of 
the course in conversational German conducted 
by Dr. Olga Janowitz on June 20. The class will 
continue to meet weekly on Fridays between 
seven and eight in the evening. 

Miss Margaret Henson is organizing a group 
of people interested in taking a Red Cross first 
aid course. Fifteen or twenty people are required 
to make a complete class, which will meet two 
hours a week for ten weeks. Anyone interested 
in taking the course should see Miss Henson in 
room 217 of the Brick Building, or Mrs. D. Mor- 
ris as soon as possible. 

The course in protozoology at the Marine Bio- 
logical Laboratory has been discontinued. 

Choral Club Rehearsals Scheduled 

Persons interested in singing good music are 
cordially invited to join the Woods Hole Choral 
Club, which will hold its first rehearsal at the 
Canteen Building of the United States Bureau of 
Fisheries on Tuesday. July 1. It is sixteen years 
since the Club held its first meeting, and during 
its existence it has provided an opportunity for 
persons connected with the laljoratory to sing 
worthwhile music under competent direction. This 
summer, as usual, the Club will rehearse for a 
concert which will be presented in the latter part 
of August. 

Prof. Ivan T. Gorokhoff, Director of Choral 
Music at Smith College, will again direct the Club 
this year. There are no dues. Rehearsals are 
held twice a week, on Tuesdays at nine o'clock 
and on Thursdays at eight. While the first re- 
hearsal will be held Tuesday, the one on Thurs- 
day this week will not be held because of the holi- 
day. 



ITEMS OF INTEREST 

RuFus H. Thomp.son, teaching assistant in 
botany at Stanford University, was injured in an 
automobile accident while he was a passenger 
en route to Woods Hole. He is convalescing in 
a hospital in Laramie, Wyoming, from a broken 
leg and a cracked shoulder blade. 



The absence of Mr. Thompson would have 
seriously interfered with the routine of the work 
of the Algae course, except for the fact that As- 
sistant Professor C.-K. Tseng of Lingnan Uni- 
versity and Mr. W. J. Gilbert of the University 
of Michigan had come to Woods Hole to continue 
their doctorate studies under the direction of Pro- 
fessor Taylor. They are devoting part of their 
time to substituting for Mr. Thompson on the 
laboratory staff. 

On July 3rd Dr. S. C. Brooks will be a guest 
lecturer before the physiology class. He will talk 
on "Some Problems in Permealjility". Dr. L. V. 
Heilbrunn lectured to the class this morning. 

The first botany seminar was held last Thurs- 
day evening; Robert H. Williams of Cornell Lhii- 
versity spoke on the "Carbohydrate Metabolism 
of the Large Brown Algae." 

Mr. Edward Chambers, son of Dr. Robert 
Chambers, married Miss Zoya Zarudnaya on 
June 3rd in New York. Miss Zarudnaya has 
been studying sculpturing in New York. They 
are expected to return to Woods Hole from New 
York in July. Mr. Chamljers is a medical student 
of New York University. 

Miss Edith Billings, who has been secretary 
in the Administration Office of the Marine Bio- 
logical Laboratory for a number of years, was 
married on June 21 to Mr. James J. Reilly, local 
contractor, at St. Joseph's Church in Woods 
Hole. 

Tennis Courts Reopen 

Both tlie mess courts and the Colas courts of 
the M.B.L. Tennis Club are now open, and it is 
expected that the beach courts will be ready for 
play in about a week. The first court was put 
into condition last Saturday, two weeks earlier 
than last year. 

Membership rates this year are $6.00 for full 
membership, and $4.00 for persons wishing to 
play only on the Colas courts. A special rate of 
$3.00 is being quoted to students taking the 
courses at the Marine Biological Laboratory. 
Guests may play on the courts for twenty-five 
cents an hour. 

The President of the Club this year is Dr. D. 
E. Lancefteld ; Dr. T. K. Ruebush is Secretary- 
Treasurer. Albert Stunkard continues as grounds- 
keeper. 



14 



THE COLLECTING NET 



[ Vol. XVI, No. 138 



THE SYMPOSIUM ON GENES AND CHROMOSOMES AT THE COLD SPRING 
HARBOR BIOLOGICAL LABORATORY 



As part of its policy of fostering a closer rela- 
tion between biology and the basic sciences, the 
Laljoratory invites each summer a group actively 
interested in a specific aspect of quantitative biol- 
ogy, or in methods and theories applicable to it. 
to carry on their work and to take part in a 
Symposium at the Lalwratory. The aim is that 
every important aspect of a given subject should 
he adequately represented, from the physical and 
chemical, as well as from the biological point of 
view. 

The Symposium this year. June 18 -July 2. 
will deal with genes and chromosomes. As a rule 
the participants will be in residence at Cold 
Spring Harbor during all of the two weeks' per- 
iod. 

F.ach (lay's program liegins at 9:30 A. M. 
When three pajiers are scheduled for the same 
dav. the third one will 1)e read at 2:15 P. M. 

STRTCTURK "I" CHROMOSOMES .XS REVEALED BY 
OPTIC.XL METHODS 

Wednesday, June 18th 

Wannke, H. E. External morphology of chromo- 
somes. 

Nebel, B. R. Structure of plant chromosomes, with 
particular emphasis on the number of chromo- 
nemata. 

Huskins, C. Leonard. The coiling of chromonemata. 

Thursday. June 19th 

Berger, C. A. Multiple chromosome complexes in 
animals and polysomaty in plants. 

S.\LIVARY GLAND CHROMOSOMES 
Metz, C. W. structure of salivary gland chromo- 
somes. 
Mazia, Daniel. Enzymatic studies of chromosomes. 

Friday, June 20th 

Painter, T. S. Chemical studies of chromosomes. 
Schultz, Jack. The evidence of the nucleoprotein 

nature of the genes. 
Cole, P., and Sutton, E. Variation in the absorption 

of ultraviolet irradiation by the bands of salivary 

gland chromosomes. 

SPONTANEOUS AND INDUCED CHANGES IN 

CHROMOSOME STRUCTURE 

Saturday, June 21st 

McClintock, Barbara. Spontaneous alterations in 

chromosome size and form in Zea. 
Kaufmann, B. P. Induced chromosomal breaks in 
Drosophila. 

Monday, June 23rd 

Sax, Karl. Effect of irradiation on plant chromo- 
somes. 

Carlson, J. Gordon. Effects of irradiation on grass- 
hopper chi-omosomes. 

Fano, U. On the analysis and interpretation of 
chromosomal changes in Drosophila. 



Tuesday, June 24th 

Delbruck, M. Biophysical analysis of chromosome 
behavior. 

MUTATIONS 

Plough, Harold H. Spontaneous mutability in Dro- 
sophila. 

Rhoades, M. M. The genetic control of mutability 
in maize. 

Wednesday, June 25th 

Demerec, M. Unstable genes in Drosophila. 
Muller, H. J. Induced mutations in Drosophila. 

Thursday, June 26th 

Stadler. L. J. Comparative studies of the genetic 
effects of X-rays and ultraviolet radiation. 

Hollaendar, Alexander. Wavelength dependence for 
mutation production with special emphasis on 
fungi. 

Gowen, John. Mutation in Drosophila, bacteria and 



rllVSICAL ASPECTS .-^ND TOOLS 

Friday, June 27th 

Schmitt, F. O. Birefringence and viscosity as a tool 
for the study of submicroscopical structures. 

Zworykin, V. K. Electi-on microscope. 

Fankuchen, I. X-ray diffraction studies of biological 
substances. 

PROPERTIES OF GIANT MOLECULES AS RELATED TO 
CHROMOSOME PROBLEMS 

Saturday, June 28th 

Mark, H. Structure and reactivity of long -chain 
molecules. 

Fruton, Joseph S. Proteolytic enzymes as specific 
agents in the formation and breakdown of pro- 
teins. 

Monday, June 30th 

Wrinch, Dorothy M. The native protein as a mega- 
molecule. 

Greenstein, Jesse P. Physical changes in thymonu- 
cleic acid induced by proteins, salts and ultravio- 
let irradiation. 

Stanley, W. M., and Knight, C. A. The chemical 
composition of different strains of tobacco mosaic 
virus. 

Tuesday, July 1st 

Claude, Albert. Particulate components of cyto- 
plasm. 

Rothen, Alexandre. Specific properties o^ proteins 
in films. 

Mirsky, A. E. Some observations on protein folding 
and unfolding. 

Wednesday, July 2nd 

Rittenberg, D. The state of the proteins in animals 
as revealed by the use of isotopes. 

CONCLUSION 
Muller, H. J. Resume and prospectives. 



June 28, 1941 ] 



THE COLLECTING NET 



15 



DIRECTORY FOR 1941 



KEY 



Laboratories 



Residence 



Botany Building Bot 

Brick Building Br 

Lecture Hall L 

Main Room in Fisheries 

Laboratory M 

Old Main Building OM 

Rockefeller Bldg Rock 

Supply Dept S 



Apartment A 

Dormitory D 

Drew House Dr 

Fisheries Residence F 

Howes Ho 

Hubbard H 

Kahler Ka 

Kidder K 

Whitman W 



MARINE BIOLOGICAL LABORATORY 



THE STAFF 

Packard, C. director, asst. prof. zool. Dept. Cancer 
Research, Columbia. 



ZOOLOGY 

Investigation 

Calkins, G. N. prof, proto. Columbia. 

Conklin, E. G. prof. zool. Princeton. 

Grave, C. prof. zool. Washington (St. Louis). 

Jennings, H. S. prof. zool. California. 

Lillie, F. R. prof. emb. Chicago. 

McCIung, C. E. prof. zool. Pennsylvania. 

Mast, S. O. prof. zool. Hopkins. 

Morgan, T. H. dir. biol. lab. California Tech. 

Parker, G. H. prof. zool. Harvard. 

Woodruff, L. L. prof, proto. Yale. 



Instruction 

Bissonnette, T. H. prof. biol. Trinity, in charge. 
Crowell, P. S., Jr. asst. prof. zool. Miami. 
Jones, E. R., Jr. prof. biol. William and Mary. 
Lucas, A. M. assoc. prof. zool. Iowa State. 
Martin, W. E. asst. prof. zool. DePauw. 
Matthews, S. A. asst. prof. biol. Williams. 
Mattox, N. T. instr. zool. Miami. 
Rankin, J. S., Jr. instr. biol. Amherst. 
Waterman, A. J. asst. prof. biol. Williams. 



EMBRYOLOGY 

Investigation (See Zoology) 

Instruction 

Ballard, W. W. asst. prof. biol. & anat. Dartmouth. 
Costello, D. P. asst. prof. zool. North Carolina. 
Goodrich, H. B. prof. biol. Wesleyan. in charge. 
Hamburger, V. assoc. prof. zool. Washington. 
Schotte, O. assoc. prof. biol. Amherst. 



PHYSIOLOGY 

Investigation 

Amberson, W. R. prof. phys. Maryland Med. 
Bradley, H. C. prof. phys. chem. Wisconsin. 
Garrey, W. E. prof. phys. Vanderbilt Med. 
Jacobs, M. H. prof. phys. Pennsylvania. 
Lillie, R. S. prof. gen. phys. Chicago. 
Mathews, A. P. prof, biochem. Cincinnati. 

Instruction 

Ballentine, R. fel. biol. Princeton. 

Fisher, K. C. asst. prof, exper. biol. Toronto. 

Kempton, R. T. prof. zool. Vassar. 

Parpart, A. K. assoc. prof. biol. Princeton, in charge. 

Prosser, C. L. asst. prof. zool. Illinois. 

Sichel, F. J. M. asst. prof. phys. Vermont Med. 

BOTANY 

Investigation 

Brooks, S. C. prof. zool. California. 
Duggar, B. M. prof. phys. & econ. bot. Wisconsin. 
Goddard, D. R. asst. prof. bot. Rochester. 
Sinnott, E. W. prof. bot. Yale. 

Instruction 

Gilbert, W. J. grad. bot. Michigan. 
Runk, B. F. D. instr. bot. Virginia. 
Taylor, W. R. prof. bot. Michigan, in charge. 
Tseng, C.-K. asst. prof. biol. Lingnan. 

INVESTIGATORS 

Allee, W. C. prof. zool. Chicago. Br 332. A 101. 
Allen, Joan OM Base. 

Alsup, F. W. grad. zool. Pennsylvania. Dr Attic. 
Amberson, W. R. prof. phys. Maryland Med. Br 109. 
Baker, Gladys E. instr. plant sci. Vassar. lib. 
Baker, H. B. prof. zool. Pennsylvania. Br 221. 
Baker, L. A. res. asst. phys. chem. Lilly Res. Labs. 

Br 319. 
Ball, E. G. asst. prof, biochem. Harvard Med. lib. 

D 314. 
Ballard, W. W. asst. prof. biol. & anat. Dartmouth. 

OM 40. 
Ballentine, R. fel. phys. Princeton. OM 2. 
Barron, E. S. G. asst. prof, biochem. Chicago. Br 

110. 
Bartlett. J. H., Jr. assoc. prof, theoretical physics. 

Illinois. OM 43. 
Benedict, D. Milton Acad. (Mass.). Br 309. 
Berger, C. A. prof, cytol. Fordham. Br 225. 
Bernheimer, A. W. grad. bact. Pennsylvania. Br 234. 

D 209. 
Bissonnette, T. H. prof. biol. Trinity. OM 28. (July 

15). 
Bliss, A. F. lect. biophysics. Columbia. Br 314. Ho 2. 
Bowen, W. J. asst. prof. zool. North Carolina. Br 

329. 
Bradford, Audrey A. Vassar. Br 227. 
Bradley, H. C. prof, physiol. chem. Wisconsin. Br 

122-A. 



16 



THE COLLECTING NET 



[ Vol. XVI, No. 138 



Brill, E. R. grad. biol. Harvard. Br 217-M. 
Brooks, Matilda M. res. assoc. zool. California. Br 

322. 
Brooks, S. C. prof. zool. California. Br 322. D 107. 
Brownell, Katherine A. res. asst. phys. Ohio State. 

Br 111. 
Budington, R. A. prof. zool. Oberlin. Br 218. (Sept. 

1). 
Bullock, T. H. fel. zool. Yale. Br 217-n. 
Cable, R. M. assoc. prof, parasit. Purdue. Br 223. 
Carothers, Eleanor res. assoc. zool. Iowa. Br 340. 
Chase, A. M. instr. biol. Princeton. Br 231. 
Claff, C. L. res. assoc. biol. Brown. Br 312. A 208. 
Clark, E. R. prof. anat. Pennsylvania Med. Br 117. 
Clark, L. B. asst. prof. biol. Union. Br 315. D 109. 
Clowes, G. H. A. dir. res. Lilly Res. Labs. Br 328. 
Cohen, L res. asst. biol. New York. Br 310. 
Cole, K. S. assoc. prof. phys. Columbia Med. Br 114. 
Colwin, A. L. instr. biol. Queens (New York). OM 

4.5. D 210. 
Colwin, Laura H. instr. biol. Vassar. OM 45. D 210. 
Commoner, B. tutor biol. Queens (New York). Br 

121. 
Compton, A. D., Jr. grad. zool. Yale. lib. 
Conger, Martha techn. N. Y. State Dept. Pub. 

Health. Br 122-B. 
Conklin, E. G. prof. biol. Princeton. Br 321. 
Copeland, M. prof. biol. Bowdoin. Br 334. 
Cornman, I. asst. zool. Michigan. K 7. Br 228. 
Costello, D. P. asst. prof. zool. North Carolina. Br 

123. D 202. 
Crawford, J. D. Milton Acad. (Mass.). Br 309. 
Crowell, S. asst. prof. zool. Miami. OM 25. 
Curtis, W. C. prof. zool. Missouri. Br 335. (Aug. 1). 
Dewey, Virginia C. res. fel. proto. Brown. OM 22. 

D 308. 
Donnellon, J. A. asst. prof. biol. Villanova. Rock 3. 
Donovan, Mary K. grad. biol. Villanova. Rock 2. 
DuBois, A. B. Milton Acad. (Mass.). Br 309. 
DuBois, E. F. prof. med. Cornell Med. Br 317. 
Duggar, B. M. prof, plant phys. & applied bot. Wis- 
consin. Br 304. (Aug. 1). 
Dumm, Mary E. grad. biol. Bryn Mawr. Br 118. D 

311. 
Dytche, Maryon grad. phys. Pittsburgh. Rock 6. 

WA. 
Dziemian, A. J. fel. zool. Pennsylvania Med. Br 204. 
Edwards. G. A. grad. asst. biol. Tufts. Br 217-0. 
Evans, T. C. asst. prof, radiol. Iowa. Br 340. D 302. 
Ferguson, F. P. asst. zool. Minnesota. Br 210. 
Fisher. K. C. asst. prof. expt. biol. Toronto. OM 

Phys. D 214. 
Forbes, J. asst. prof. biol. Fordham. Br 225. 
Frasier, Doris A. instr. biol. Russell Sage. OM 44. 
Frisch, J. A. prof. biol. Canisius (Buffalo). OM 39. 
Gabriel, M. L. asst. zool. Columbia. Br 314. 
Garrey, W. E. prof. phys. Vanderbilt Med. Br 215. 
Gates. R. R. prof. bot. London (Ont.). lib. 
Gilbert, W. J. grad. bot. Michigan. Bot. Dr 6. 
Gilman, L. C. grad. zool. Hopkins. OM 36-b. 
Glancy, Ethel A. tutor biol. Queens (New York). 

OM 36. 
Goldin, A. grad. zool. Columbia. Br 313. 
Goodrich, H. B. prof. biol. Wesleyan. Br 210. D 204. 
Goulding, Helen J. gi-ad. phys. Toronto. 
Grave, C. prof. zool. Washington (St. Louis). Br 327. 
Groupe, V. grad. immun. Pennsylvania. Br 234. 
Guttman, Rita instr. phys. Brooklyn. Br 110. 
Haas, W. J. M.I.T. Br 116. 

Hager, R. P. res. asst. zool. Pennsylvania. Rock 2. 
Hamburger, V. assoc. prof. zool. Washington (St. 

Louis). Br 303. 
Hamilton, Pauline G. grad. res. asst. zool. Pennsyl- 
vania. Br 220. 
Harris, D. L. instr. zool. Pennsylvania. Br 217-e. 



Hartman, F. A. prof. phys. Ohio State. Br 111. D 

218. 
Harvey, E. N. prof. phys. Princeton. Br 116. 
Harvev, Ethel B. indep. invest, zool. Princeton. Br 

116. 
Hassett, C. grad. asst. zool. Hopkins. Br 315-B. 
Hayashi, T. grad. asst. zool. Missouri. Br 110. K 21. 
Hayes, E. R. grad. asst. anat. Ohio State. OM 46. 
Haywood, Charlotte assoc. prof. phys. Mt. Holyoke. 

Br 335. A 207. 
Heilbrunn, L. V. assoc. prof. zool. Pennsylvania. Br 

220. 
Hendley, C. D. grad. asst. biophysics. Columbia. Br 

314. 
Henry, R. J. Pennsylvania Med. OM Phys. Ho 6. 
Henson, Margaret fel. biol. New York. Br 217-E. 

H 9. 
Hibbard, Hope prof. zool. Oberlin. Br 218. 
Hickson, Anna K. res. chemist. Lilly Res. Labs. Br 

319. 
Hill, S.E. prof. biol. Russell Sage. OM 44. 
Hinchey, M. Catherine grad. biol. Temple. Br 214. 

(Aug. 9). 
Hohwieler, H. J. grad. asst. zool. Washington (St. 

Louis). Br 207. 
Hopkins, D. L. prof. biol. Mundelein (Chicago). Bot 

5. 
Houck, C. R. fel. phys. Princeton. Br 231. 
Howe, H. E. ed. Indus. & Engineering Chem. Br 203. 
Hunninen, A. V. prof. biol. Oklahoma City. Br 217-k. 
Hutchens, J. O. grad. zool. Hopkins. Br 325. 
Hyman, C. res. asst. biol. New York. Br 343. 
lUick, J. T. assoc. prof. zool. Syracuse. OM 36-h. 

Dr 2. 
Jacobs, M. H. prof. gen. phys. Pennsylvania. Br 205. 
Jandorf, B. J. res. asst. biochem. Lilly Res. Labs. 

Br 333. 
Janowitz, Olga instr. Potomac School (Washington, 

D. C). L 24. D 318. 
Jenkins, G. B. prof. anat. George Washington. Br 

122-D. 
Jones, E. R., Jr. prof. biol. William & Mary. OM 33. 

D 206. 
Katzin, L. I. jr. biologist. U. S. Pub. Health (Cin- 
cinnati). Br 217-g. 
Kempton, R. T. prof. zool. Vassar. OM 3. 
Knowlton, F. P. prof. phys. Syracuse Med. Br 226. 
Krahl, M. E. res. chemist Lilly Res. Labs. Br 333. 

A 301. 
Lansing, A. I. asst. zool. Indiana. Rock 6. Dr Attic. 
Lerner, E. M., II. Harvard. Br 122. 
Levin, E. grad. phys. Ohio State. Br 111. 
Lewis. Lena A. res. asst. phys. Ohio State. Br 111. 
Liebmann, E. res. fel. anat. Tulane Med. Bot. 1. 
Lillie, F. R. prof. emb. Chicago. Br 101. 
Lillie, R. S. prof. phys. Chicago. Br 326. 
MacHaffie, R. grad. biol. Columbia. Br 110. 
Markell, E. K. grad. zool. California. Br 217-m. 
Marmont, G. res. assoc. phys. Columbia Med. Br 114. 
Marsland, D. A. asst. prof. biol. New York. Br 344. 
Martin, Rosemary D. C. dem. phys. Toronto. OM 

Phvs. 
Martin, W. E. asst. prof. zool. DePauw. OM 31. 
Mast, S. O. prof. zool. Hopkins. Br 329. 
Mathews, A. P. prof, biochem. Cincinnati. Br 341. 
Mattox, N. T. instr. zool. Miami (Ohio). OM 32. 

(July 15). 
Mavor, J. W. prof. biol. Union. Br 315. 
McClung, C. E. prof. zool. Illinois. Br 219. 
McNutt, W. S., Jr. asst. biol. Brown. Br 331. 
Melkon, B. grad. zool. Pennsylvania. OM 36-e. Ho 1. 
Menkin, V. asst. prof. path. Harvard Med. L 27. 
Messer, Anne C. techn. Harvard Sch. Pub. Health. 

Br 110. 



June 28, 1941 ] 



THE COLLECTING NET 



17 



Metz, C. B. fel. emb. California Tech. 

Meyerhof, O. H. res. prof, biochem. Pennsylvania 

Med. Br 108. 
Miller, J. A. instr. anat. Michigan. Br 228. 
Milne, L. J. assoc. prof. biol. Randolph-Macon. Br 

233. D 205. 
Miriam Elizabeth (Sister) assoc. prof. biol. Chest- 
nut Hill (Phila.). Rock 3. 
Mitchell. P. H. prof. biol. Brown. Br 331. 
Molnar, G. W. instr. zool. Miami (Ohio). OM 36-f. 
Molter, J. A. grad. zool. Pennsylvania. OM Base. 
Moog, Florence grad. zool. Columbia. Br 314. H 9. 
Moore, W. G. instr. biol. Loyola. Br 110. 
Morgan, T. H. prof. biol. California Tech. Br 320. 
Morrill, C. V. assoc. prof. anat. Cornell Med. Br 317. 
Mullins, C. P. instr. phys. Rochester. Br 322. 
Nachmansohn, D. res. fel. phys. Yale Med. Br 110. 

D 112B. 
Navez, A. E. tutor. Milton Academy (Mass.). Br 309. 
Nelson, G. H. Milton Acad. (Mass.). Br 309. 
O'Brien, J. P. grad. zool. Hopkins. Bot 1. 
Oesterle, Paul D. (Sister) instr. biol. Chestnut Hill 

(Phila.). Rock 3. 
Olson, M. instr. zool. Minnesota. Br 217-n. (Aug. 1). 
Osbom, C. M. instr. anat. Ohio State. OM 46. 
Osterhout, W. J. V. mem. Rockefeller Inst. (New 

York). Br 207. A 203. 
Osterud, Dorothy W. res. asst. zool. New York. Br 

232. 
Osterud, K. L. grad. asst. zool. New York. L 21. 
Oxford, A. E. fel. biochem. Rockefeller Found. Br 

342. D 309. 
Parker, G. H. prof. zool. Harvard. Br 213. A 308-9. 
Parmenter, C. L. prof. zool. Pennsylvania. Br 221. 

D 201. 
Parpart, A. K. assoc. prof. phys. Princeton. OM 2. 
Phelps, Lillian A. asst. prof. biol. Washburn. OM 

Base. 
Philips, F. S. fel. biol. Yale. Br 110. (July 15). 
Plough, H. H. prof. biol. Amherst. Br 330. 
Pollister, A. W. prof. zool. Columbia. Br 313. (Sept. 

1). 
Prosser, C. L. asst. prof. zool. Illinois. OM 7. 
Rankin, J. S.. Jr. instr. biol. Amherst. OM 24. 
Recknagel, R. grad. zool. Pennsylvania. OM Base. 

Ho 3 
Ris. H. asst. zool. Columbia. Br 314. Dr 5. 
Rona, Elizabeth fel. geophysics. Carnegie Inst. 

(Washington). Br 108. 
Ronkin, R. R. grad. zool. California. Br 322. K 24. 
Rothstein, A. grad. asst. biol. Rochester. Br 322. 
Ruebush, T. K. instr. zool. Yale. L 26. Dr 7. 
Runk, B. F. D. instr. bot. Virginia. Bot 26. D 110. 
Sayles, L. P. asst. prof. biol. C.C.N.Y. Rock 6. D 304. 
Schaeffer, A. A. prof. biol. Temple. Br 214. D 313. 
Schaffel, M. res. asst. biol. Pittsburgh. Rock 7. 
Schechter, V. asst. prof. biol. C.C.N.Y. Br 315-a. D 

102. 
Scott, A. asst. prof. biol. Union. 
Scott, Florence M. (Sister) prof. biol. Seton Hill 

(Greensburg, Pa.). Br 225. 
Shapiro, H. instr. phys. Vassar. Br 227. 
Shaw, Myrtle senior bact. N. Y. State Dept. Health. 

Br 122-B. D 303. 
I Shelanski, L. grad. zool. Pennsylvania. OM Base. Dr 
j 15. 

Shelden, F. F. instr. phys. Ohio State. Br 111. K 24. 
Sichel, El.sa K. head sci. dept. State Normal Sch. 

(Johnson, Vt.). OM 4. K 8. 
I Sichel, F. J. M. asst. prof. phys. Vermont Med. OM 

1 ''■ 

j Slaughter, J. C. grad. asst. zool. Iowa. Br 340. 

\ Slifer, Eleanor H. asst. prof. zool. Iowa. Br 217-a. 

1 D 203. 



Smith, J. A. instr. phys. & Pharmacol. Chicago Med. 

Br 6. 
Solberg, A. N. asst. prof. biol. Toledo, lib. (Aug. 3). 
Speidel, C. C. prof. anat. Virginia. Br 106. D 315. 
Steinbach, H. B. asst. prof. zool. Columbia. Br 313. 
Stern, K. G. res. asst. prof, physiol. chem. Yale Med. 

Br 110. (Aug. 1). 
Stokey, Alma G. prof. bot. Mt. Holyoke. Bot 1. 
Stowell, R. E. res. asst. Barnard Hospital (St. 

Louis), lib. 
Stunkard, H. W. prof. biol. New York. Br 232. 
Sturtevant, A. H. prof. biol. California Tech. Br 126. 
Tashiro, S. prof, biochem. Cincinnati. Br 341. 
Taylor, W. R. prof. bot. Michigan. Bot 25. 
TeAVinkel, Lois E. asst. prof. zool. Smith. Br 217-b. 

A 205. 
Thivy, Francesca grad. bot. Michigan. Bot. 
Trinkaus, J. P. grad. zool. Columbia. OM 41. 
Troedsson, Pauline H. grad. zool. Columbia. Br 314. 
Trombetta, Vivian V. instr. bot. Smith. Bot 1. 
Tseng, C.-K. asst. prof. bot. Lingnan (China). Bot 1. 

Dr 6. 
Wald, G. instr. biol. Harvard. Br 318. 
Walker, R. instr. biol. Rensselaer. OM 38. 
Warner, E. N. instr. zool. Ohio State. OM 36. 
Waterman, A. J. asst. prof. biol. Williams. OM 26. 

(July 20). 
Watterson, Ray asst. biol. Hopkins. OM 41. 
Weisiger, J. R. grad. fel. physiol. chem. Hopkins. 

Br 224. D 111. 
Wenrich, D. H. prof. zool. Pennsylvania. Br 219. 

(Aug. 1). 
Whiting, Anna R. guest invest, zool. Pennsylvania. 

Rock 2. 
Whiting, P. W. assoc. prof. zool. Pennsylvania. Rock 

2. 
Wichterman, R. asst. prof. biol. Temple. Br 217-h. 
Wiercinski, F. J. res. asst. zool. Pennsylvania. Br 

220. K 7. 
Wilbur, K. M. res. assoc. biol. New York. Br 342. 

K 12. 
Willier, B. H. prof. zool. Hopkins. Br 324. A 302. 
Wilson, W. L. grad. zool. Pennsylvania. OM Base. 
Wolf, E. A. assoc. prof. biol. Pittsburgh. Rock 7. 
Wolf, Opal M. asst. prof. zool. Goucher. Br 122-C. 

A 206. 
Woodruff, L. L. prof, proto. Yale. Br 323. 
Woodward, A. A. grad. asst. biol. New York. L 21. 

Dr 3. 
Woodward, A. E. asst. prof. zool. Michigan. (Aug. 

20). 
Wrinch, Dorothy lect. chem. Hopkins, lib. 
Yntema, C. L. asst. prof. anat. Cornell Med. Br 317. 

D 310. 
Young, W. C. assoc. prof, primate biol. Yale. lib. 

(Aug.). 
Zarudnaya, Katya Radcliffe. OM 36. H 6. 
Zinn. D. J. grad. zool. Yale. OM 38. 
Zorzoli, Anita asst. zool. Columbia. OM 36. H 7. 
Zweifach, B. W. res. assoc. biol. New York. Br 310. 



STUDENTS 

Abbott, C. C. Haverford. bot. 

Algire, G. H. gi-ad. anat. Maryland Med. phys. 

Bayard, Ellen B. U. of Connecticut, bot. H 8. 

Bergstrora, W. H. Amherst, emb. 

Birmingham, L. W. Rochester, emb. Dr 3. 

Bull, Nancy B. Wellesley. bot. H 2. 

Burns, J. E., Jr. grad. asst. biol. Wesleyan. phys. 

Coe, F. W. Ohio Wesleyan. phys. Ho 6. 

Cole, R. M. teach, fel. biol. Harvard, emb. Ka 3. 



18 



THE COLLECTING NET 



[ Vol. XVL No. 138 



Covalla, Miriam J. instr. biol. Seton Hill (Greens- 
burg, Pa.), emb. H 1. 

Crumb, Cretyl Wellesley. emb. H 1. 

Deyrup, Ingrith J. Barnard, phys. 

Egan, R. W. res. asst. biol. Canisius (Buflfalo, N. Y.). 
phys. Dr 2. 

Eldred, E. asst. biol. Northwestern, emb. Dr 3. 

Enzenbacher, Jean A. Smith, bot. W B. 

Frantz, Ruth E. Elmira. bot. D 307. 

Gibbs, Elizabeth Wheaton. emb. W C. 

Gordon, H. T. teach, fel. biol. Harvard, phys. Ho 1. 

Green, J. W. Davis-Elkins. phys. 

Gregg, J. R. asst. biol. Alabama, phys. 

Gross, J. B. De Pauw. emb. 

Harrison, R. W. grad. asst. biol. Wesleyan. phys. 
K 7. 

Hartmann, J. F. asst. zool. Cornell, phys. Dr 1. 

Hasse. G. W. grad. asst. zool. Illinois, emb. 

Hendricks, Anne L. Cincinnati, emb. 

Henry, Jane E. Gettysburg, phys. H 3. 

Hollister, Patricia Sarah Lawrence, emb. 

Hopkins, Marjorie G. grad. asst. biol. Mt. Holyoke. 
emb. H 7. 

Jacobs, J. teach, fel. biol. New York. phys. Dr 1. 

Josephson, N. Wesleyan. emb. K 5. 

Karczmar, A. G. grad. biol. Columbia, emb. 

Katz, Elaine J. Goucher. bot. D 307. 

Keezer, W. S. Indiana, phys. K 15. 

Kezer, L. J. instr. biol. State Teachers (Newark, N. 
J.), emb. K 9. 

Kieffer, R. F., Jr. Franklin and Marshall, emb. Ho 2. 

Kirkpatrick, Elizabeth M. Connecticut College, emb. 

Lambie, M. W. Harvard, emb. 

Lein, J. C.C.N.Y. phys. Ka 22. 

Lerner, Eleanor D. grad. biol. Brooklyn, emb. W D. 

Leuchs, Augusta V. H. A. grad. biol. Radcliffe. bot. 
H 2. 

Linthicum, Anne H. Goucher. emb. D 106. 

Lorenz, P. B. Swarthmore. phys. Ho 3. 

Martin, R. G. Harvard, emb. Dr 15. 

Martin, T. S. Oberlin. emb. K 15. 

Medlicott, Mary grad. asst. biol. Mt. Holyoke. emb. 
H 7. 

Morgan, T. W. Washington and Jefferson, emb. Dr 
10. 

Mothes, Arlene M. Massachusetts State, emb. W E. 

Muchmore, W. B. Oberlin. emb. K 7. 

Muir. R. M. grad. bot. Michigan, bot. Dr 6. 

Perkins, Patricia J. grad. chem. Cincinnati, phys. 
W F. 

Plough, I. C. Amherst, emb. 

Power, M. E. grad. asst. zool. Yale. emb. Ka 2. 

Regnerv, D. C. lab. instr. biol. Stanford, emb. K 9. 

Reiner, E. R. lab. asst. biol. Alabama, emb. 

RoUason, H. D., Jr. grad. asst. biol. Williams, phys. 
Dr 7. 

Rosenblum, E. D. Brooklyn, phys. K 10. 

Rovle. Jane G. grad. biol. Bryn Mawr. emb. 

Salvin, S. B. grad. asst. biol. Harvard, bot. Ka 3. 

Saunders, Grace S. teach, fel. biol. New York. emb. 
H 9. 

Schepartz, B. Ohio Wesleyan. phys. Dr 5. 
Schlosser, Adele P. Vassar. phys. 
Smith. F. B. prof. biol. Florida, bot. 
Smithcors, J. F. Rutgers, emb. Ka 2. 
Stanton. Constance L. Bryn Mawr. bot. W G. 
Stenger, F. R. prof. biol. St. Mary of the Lake Sem- 
inary (Mundelein, 111.), bot. 
Stern, J. R. res. asst. med. Toronto, phys. K 6. 
Svihla, G. res. asst. zool. Illinois, emb. Ho 2. 
Thorne, R. F. Dartmouth, bot. Ka 21. 
Timanus, Alice L. Converse, emb. D 106. 
Tuttle, L. Constance grad. asst. biol. Mt. Holyoke. 
phys. W H. 



Waldron, Jacqueline M. American, bot. H 3. 
Wasserman, E. asst. biol. Wesleyan. emb. K 5. 
Weiner, H. M. Harvard, phys. Dr 1. 
Williams, R. H. instr. bot. Cornell, bot. D 208. 
Zimmerman, G. L. Swarthmore. phys. Ho 3. 
Zingher, J. M. C.C.N.Y. emb. K 10. 

OFFICE OF ADMINISTRATION 

Crowell, Polly L. asst. to bus. mgr. 
MacNaught, F. M. bus. mgr. 
Packard, C. director. 
Reilly, Edith Billings sec. 
Whitcomb, Mary sec. W I. 



LIBRARY 

Lawrence, Deborah sec. 
Montgomery, Priscilla B. librarian. 
Rohan, Mary A. asst. 
Thombs, S. Mabell asst. WF. 



MUSEUM 
Gray, G. M. curator emer. 



DEPARTMENT OF CHEMICAL SUPPLIES 
AND SCIENTIFIC APPARATUS 

Chemical Room 

Ballard, K. C. teach, sci. Lawrence H.S. (Falmouth). 
Beazley, Dorothea B. Falmouth. 
Cherry, Bettv Tufts Med. WD. 
Hatch, Milford H. Brown. 
Heimberg, Felix Harvard Med. Dr 3. 
Smith, J. A. prof. biol. & Pharmacol. Chicago Med. 
Thompson, John Lawrence H. S. 

Weisiger, J. R. fel. physiol. chem. Hopkins. Br 224. 
D 111. 

Apparatus and Technical Service 

Boss, L. F. techn. Br 6. 

Bridgman, Jane grad. biol. Harvai'd. photographer. 

H. 
Graham, J. D. Pennsylvania, glass blower. Br 22. 
Lefevre. Dorothy E. sec. Br 1. 
Liljestrand, Robert S. Br 3. 
Pond, S. E. tech. mgr. Br 1-3. 

EXPERIMENTAL RADIOLOGY 

Failla, G. Memorial Hosp. Br 307-8. 

Little, E. P. instr. Phillips Exeter. Br 307-8. 



MAINTENANCE 

Alper, C. janitor. Dr Attic. 

Bacchus, S. janitor. Ka 1. 

Blanchard, L. A. janitor. 

Cannon, F. janitor. 

Fink, F. janitor. Ka 4. 

Genther, T. janitor. Ka 4. 

Hemenway, W. C. carpenter. 

Kahler, R. S. asst. 

Larkin, R. janitor. 

Larkin. T. E. supt. Br 7. 

Roth, P. janitor. Ka 1. 

Spier, R. janitor. Ka 4. 

Tawell, T. E. head janitor. 

Taylor, R. night mechanic. Dr Attic. 



June 28, 1941 ] 



THE COLLECTING NET 



19 



Travis, R. F. mail. 
Trinkaus, W. janitor. Ka 1. 
Wynn, J. fireman. 

SUPPLY DEPARTMENT 

Bissonnette, J. animal house. 

Bulmer, Gladys teacher H.S. (Philadelphia), collec- 
tor. 

Carlson, B. C. Phillips Exeter, collector. 

Crowell, Ruth S. sec. 

Egloif, F. collector. 

Gilbert, W. J. Michigan, hot. collector. Dr 6. 

Glass, B. A & M College (Texas), collector. Ho 9. 

Goodrich, A. animal house. Ho 5. 

Gray, M. B. collector. 

Harman, Grace sec. WH. 

Hilton, A. M. collector. 

Kahler, W. E. collector. 

Kyllonen, A. Harvard, collector. Ho 4. 

Kyllonen, D. collector. Ho 4. 

Leathers, A. W. head shipper. 

Lehy, G. collector. 

Leonard, E. collector. Ho 4. 

Melnnis, J. mgr. 

Metcalf, W. G. Oberlin. collector. Ho 5. 

Parker, J. collector. 

Poole, Margery hot. collector. 

Ross, F. collector. 

Sheldon, D. collector. Ho 4. 

Talbert, J. D. Columbia (Mo.), collector. Ho 5. 

Wamsley, F. W. supervisor of schools (Charleston), 
preparator. 

Young, E. Worcester Acad, collector. 

THE BIOLOGICAL BULLETIN 

Boyden, Louise E. ed. asst. Br 120. 
Redfield, A. C. managing ed. Br 120. 

THE JOURNAL OF INDUSTRIAL AND 
ENGINEERING CHEMISTRY 

Anderson, Stella B. sec. Br 203. 
Bruff, Eleanor G. sec. Br 203. 
Gordon, Gladys sec. Br 203. 
Howe, H. E. editor. Br 203. 
Martenet, Dorothy sec. Br 203. 
Newton, Helen K. ms. ed. Br 203. 
Parkinson, Nellie A. sec. Br 203. 



THE COLLECTING NET 

Cattell, W. managing ed. Scientific Monthly. 
Gorokhofif, B. I. Yale. 
Woodring, Judy Maryland. 

WOODS HOLE OCEANOGRAPHIC 
INSTITUTION 

Barnes, C. H. physical oceanographer, U. S. Coast 

Guard. 302. 
Bumpus, D. F. biol. techn. 108. 
Clarke, G. L. marine biologist. 107. 
Dooley, D. asst. submarine geol. 206. 
Ewing, W. M. assoc. submarine geol. 102. 
Hagelbarger, D. fellowship holder. 103. 
Hamilton, W. asst. submarine geol. 212. 
Irving, L. physiol. investigator. 315. 
Iselin, C. O'D. director. 114. 
Ketchum, B. H. assoc. marine biol. 203. 



Lee, R. fel. 310. 

Montgomery, R. B. physical oceanographer. 208. 

Myers, Dr. & Mrs. E. H. visiting invest. 308. 

Orr, E. chemical techn. 207. 

Osborn, C. M. biol. invest. 111. 

Parker, G. H. biol. invest. 111. 

Phleger, F. geol. invest. 212. 

Rakestraw, N. W. chem. oceanographer. 109. 

Redfield, A. C. assoc. marine biol. 309. 

Renn, C. E. assoc. marine bact. 211. 

Schaliek, W. visiting invest. 306. 

Sears, Mary planktonologist. 305. 

Seiwell, H. R. physical oceanographer. 301. 

Soule, F. M. principal physical oceanographer, U. S. 

Coast Guard 303. 
Stetson, H. C. submarine geol. 212. 
Von Brand, Th. chem. invest. 110. 
Waksman, S. A. marine bact. 211. 
Weiss, C. M. bacteriol. techn. 201. 
Whiteley. G. fel. 305. 
Whitney, J. chem. asst. 109. 
Woodcock, A. J. oceanographic techn. 207. 
Zabor, J. W. fel. 109. 

Olfice of Administration 

Bird, Ethelyn sec. to bus. mgr. 113. 
Phillips, H. F. sec. to the dir. 112. 
Schroeder, W. C. bus. mgr. 113. 

"Atlantis" 

Backus, H. first engineer. 
Cook, H. second engineer. 
Kelley, T. N. first officer. 
Mandly, H. second officer. 
McMurray, F. S. captain. 



"Anton Dohrn" 



Atheran, E. captain. 
Daggett, R. engineer. 
Poole, S. mate. 



Buildings and Grounds 

Condon, W. asst. to supt. 
Schroeder, W. supt. 

U. S. BUREAU OF FISHERIES 

SCIENTIFIC STAFF 

Algire, Dorothy H. asst. biol. U.S.B.F. 122. F 27. 

Bliss, C. I. indep. invest. 119. F 43. 

Galtsoff, Eugenia assoc. zool. George Washington. 

123. F 23-24. 
Galtsoff, P. S. biol. U.S.B.F. acting director. 118. F 

23-24. 
Pupchick, Anna sec. 118. F 30. 

BUILDINGS AND GROUNDS 

Armstrong, J. apprentice fish culturist. 
Bellinger, H. H. fireman. 
Collins, E. J. apprentice fish culturist. 135. 
Conklin, P. fireman. Hatchery 137. 
Goffin, R. A. superintendent. 117. F. 
Hamblin, R. P. apprentice fish culturist. 
Howes, E. S. coxswain. 116. 
Jackson, R. fish culturist. 
Lowey, J. engineer. 



20 



THE COLLECTING NET 



[ Vol. XVI, No. 138 



Immunity against 
Animal Parasites 



This new book by James T. Culbertson. 
Assistant Professor of Bacteriology in 
Columbia University, is designed to pro- 
\\t\ij Ijoth the fundamental principles of 
immunity to parasitic infection and a re- 
view of the literature of the last twelve 
years. It will be published in the early 
fall. Price, $3.50 



Protozoa in 
Biological Research 



Written by twenty leading specialists, 
under the editorship of Gary N. Calkins 
and Francis M. Summers, this book 
summarizes the latest developments and 
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together the most recent information 
about the structure and bodily functions 
of the protozoa and their relations with 
their environment and analyzes it for its 
significance in general Ijiological re- 
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Genetics and the 
Origin of Species 



In ils original form this work by Theo- 
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new. up-to-date edition takes into con- 
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June 28, 1941 ] 



THE COLLECTING NET 



21 




GOLD SEAL 



.over 



WE ARE PROUD to be able to offer you 
cover glasses entirely made in the United 
States. Formerly GOLD SEAL cover glasses 
were cut, gauged, sorted, and packed in our 
ow^n workshop from imported glass. Now, 
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are proud of this accomplishment. 

American made GOLD SEAL Cover 
Glasses are at least equal in NON-CORRO- 
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July 14th to 26th. 



U. S. A. 



careful laboratory testing and control . . . 
Physically, the glass is practically free 
of stria, bubbles, and curvature. 

Prices have been kept low v/hen it 
is considered that the cost of domestic 
production is many times the import prices. 
See page 22 of our new Catalog No. 1Q2 
CN for prices — if you haven't a copy 
please write for one on your letterhead. 



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CO., 44 East 23rd Sr. ,.„.„, 
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The above Centrifuges will accommodate six 15 ml. 
round bottom or taper bottom test tubes. Micro and 
semimicro tubes (5 ml. to 0.5 ml.) can be accommodated 
by purchasing extra shields, prices from 50c to 75c 
each. No adapters are required. 

Other ADAMS CENTRIFUGES and laboratory 
supplies are described in cur new Catalog No. 
102CN. If you do not already have a copy write 
for one on your letterhead please. 

See our Exhibit in the Old Lecture Hall, 
July 14th to 26th. 




5000 R. p. M. on D. C. 

with underguard No. CT-1055 
with six 15 ml. tubes loaded. 
4200 R.P.M. on A. C. with same load. 

ADAMS ANGLE CENTRIFUGE 

These centrifuges offer important advantages over 
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22 



THE COLLECTING NET 



[ Vol. XVL No. 138 




New! JUSTRITE Replaceable Blade Scalpel 



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BIOLOGICAL SLIDES 

Credit for the leadership of Turto.x microscope slides in American schools goes to the 
highly trained individuals who make these slides in the Turtox Slide Laboratory. These men 
and w I mien lia\ e s])ecialized in their particular fields and devote their entire efforts to just 
one end — the pre])aratiun of the best possilile biological slides. 

MITOSIS SLIDES 

111 addition to the usual mitosis material such as onion, Tradpscantia, Ascaris, crayfish, Amby- 
stonia and others commonly used for the study of mitosis, we offer our slides of whitefish blastula. 
Ercnj slide shows every stage of mitosis. Many teachers now usiug them consider these preparations 
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June 28, 1941 ] 



THE COLLECTING NET 



23 




Shown above is Spencer Stereoscopic Microscope No. 26 with a new Spencer lamp attached. 

Spencer has improved 
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Your very first experience with the 
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A wide range of magnification is avail- 
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Notable mechanical improvements have 
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Sec exhibit oj eoinplele Spencer eqitipiiiciit in Canlecii IJidUliiig, 
Bureau oj Pixlteries. June 30 to Ju!y 11 . 



THE COLLECTING NET 



[ Vol. XVI, No. 138 




Sand— Symbol of Optical Independence 



BY itself, only a handful of sand — fine, pure, 
white crystals of quartz from a Pennsylvania 
hillside. But, blended with boron, sodium, barium, 
lead, phosphorus and other elements — fused and 
fined at white heat — cooled, sorted, annealed and 
selected — it becomes optical glass, one of the basic 
indispensable materials of national defense — and 
of modern civilization. 

Thirty years ago America was wholly dependent 
on Europe for a supply of glass for optical instru-_ 
ments. But before the first World War had cut off 
that source, Bausch & Lomb scientists, at Roch- 
ester, New York, were at work on the development 
of a glass-making technique. By 1918, glass to fill 
the vital needs of optical manufacturing in the 



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Vol. XVI, No. 2 



SATURDAY, JULY 5, 1941 



Annual Subscription, $2.00 
Sinprle Copies. 30 Cents. 



PROTOPLASMIC STREAMING 

Dr. Douglas A. Marsland 

Assisfaiif Professor of Biology, 
irashiiigton Square College, A^. Y. Unkfersity 

High hydrostatic pressure, the type of pressure 
which reaches great intensity in the oceanic 
depths, has been applied artificially to various 
living tissues and has proved 
a useful tool for analyzing 
some of the reactions which 
take place in the protoplasma 
of living cells, — reactions 
which provide the energy for 
the characteristic activity of 
the cell. One type of activity, 
namely protoplasmic stream- 
ing, appears to be especially 
sensitive to the effects of pres- 
sure. The streaming of the 
protoplasm of various Amoe- 
boid cells during active loco- 
motion, the protoplasmic cur- 
rents which occur at the time 
that animals are dividing, the 
cyclic flow of protoplasm 
which is characteristic of so 
many plant cells, and the ebb 
and flow of pigment granules 
back and forth in the fine 



% 1. (Calfnacr 



TUESDAY, July 8, 8:00 P. M. 

Seminar: Physiological seminar in 
which the following investiga- 
tors will take part: A. J. Dzie- 
mian, Rita Guttman and Herbert 
Shapiro. 



FRIDAY, July 11, 8:00 P. M. 

Lecture: "The Integration of Neu- 
rology and Psychology," Dr. K. 
S. Lashley, Research Professor 
of Neuropsychology, Harvard 
University. 



HOW FEATHERS ARE MADE 

Dr. Frank R. Lillie 

Professor of Embryology, Emeritus, 

University of Chicago 

I. Introdiictiou 
If this question could be fully answered within 
the limits of present scientific principles and 
_ methods we would have much 
deeper insight into problems 
of physiology, genetics and de- 
velopment. But our knowl- 
edge is still very incomplete ; 
and of the httle that we know 
I shall be able to tell you only 
a part of my own work, done 
originally with the aid of Dr. 
Alary Juhn and recently of 
Air. Hsi Wang. 

Each feather is an autobio- 
graphical record written in the 
course of its life history — a 
sort of diary, in fact. To read 
it one needs a Rosetta Stone, 
and this is furnished by study 
of its growth and development. 
By plotting rates of axial 
growth with reference to the 
location of simultaneous reac- 
tions in the germ (isochrones) 



branches which radiate out from the pigment cells it is possilale to relate any definitive part of the 
of the skin of many fishes, — all of these forms feather to its place and time of origin, and hence 
of streaming are re- {Continued on page 31) to the conditions of determination when these are 



TABLE OF CONTENTS 



How Feathers are Made, Dr. Frank R. Lillie.... 25 

Protoplasmic Streaming, Dr. Douglas A. 

Marsland 25 

Notes on Eulima oleacea Embryology, George 

M. Gray (cont.) 29 

Grafting of Limbs in Place of the Eye in 

Amblystoma, Dr. Jean Piatt 32 



M.B.L. Department of Chemical Supplies and 

Scientific Apparatus 33 

Children's School of Science 34 

Items of Interest 35 

Physiology Class Notes 36 

Botany Class Notes 36 

Embryology Class Notes 37 

A.B.C. of Woods Hole 38 




^ o 

<; us 

K X! 

<; a 

o £ 



5 tm 



July 5, 1941 ] 



THE COLLECTING NET 



27 



known. By virtue of these properties, for in- 
stance, relations between rates of growth and 
thresholds of reaction have been established, and 
also between gradients of threshold and structure 
and pattern in the individual feathers. 

Regenerating feathers of the fowl are excellent 
material for study. They can be made available 
at any time by plucking, and during most of the 
period of regeneration they are extremely sensi- 
tive indicators to alterations of physiological con- 
dition, both natural and such as may be induced 
by controlled experiments. 

Experiments with reactions of saddle and neck 
hackle feathers of Brown Leghorn males to injec- 
tions of thyroxin illustrate sensitiveness of reaction 
of growing feathers. (Lantern slides). A single in- 
jection of 0.5 mg. in a bird weighing 1800 grams 
produces a spindle-shaped black mark adjacent to 
the rhachis, the length of which indicates the du- 
ration of reaction, and the form of -which indi- 
cates absorption to the center of the spindle and 
excretion thereafter of the excess thyroxin. The 
threshold of reaction is least immediately next to 
the rhachis and becomes increasingly higher to- 
wards the apex of the barbs. Single injections 
of 1.0 mg, 1.5 mg, 5.0 mg, etc., produce succes- 
sively broader areas until the margin of the 
feather is reached with 5.0 mg. There is thus 
a gradient of threshold along the axis of each 
barb to thyroxin. 

The stated action of thyroxin is on the melano- 
blasts, which rapidly grow into melanophores, 
producing pigmentation otherwise absent. 

This method may be used also to produce in- 
teresting patterns, as illustrated by neck hackle 
feathers in which successive injections of 1.0 mg. 
of thyroxin every seventh day produces a repeated 
pattern, and 1.5 mg. every sixth day produces a 
similar repeated pattern but broader and closer. 

//. General 

In order to give an account of the experiments 
which form the principal subject of this lecture 
it is necessary to introduce a short account of the 
anatomy and development of the feather. Feathers 
from different tracts of one bird differ from each 
other much as animals of two different species do. 
The account therefore specifies the feathers of the 
breast and saddle tracts of the Brown Leghorn 
male. Study of particulars is necessary in order 
to reach sound general conclusions. 



1. Anatomy 

The feather is not formed l)y budding and 
liranching as are, for instance, "feathery" hydroids 
(such as Pennaria), but is composed of parts 
separately formed and secondarily assembled. It 
can therefore be taken apart like a machine. These 
parts are the shaft (rhachis), the barb stem, and 
the barbules. The barbules do not grow out of 
the Imrb stem. The barbs do not grow out of 
the rhachis. 

The rhachis is formed of a central and two 
lateral components. The lateral components carry 
the barbs and are, indeed, composed of the union 
of modifications of the bases of barbs called the 
"barb petioles" which are bound together by long 
keratin fibers originating within the petioles. (Il- 
lustrated b}' lantern slides.) 

2. Sketch of Normal Development 

The feather develops from a "papilla" situated 
at the bottom of a follicle six to eight mm. deep 
in the case of the breast feather opening on the 
surface of the skin. It emerges from the follicle 
in the form of a cylinder covered by a stiff sheath 
of keratin. As the feather grows the barbs and 
rhachis Ijurst through the sheath and extend far 
beyond it. The base of the growing feather is 
always in the form of a cylinder. 

A longitudinal section of the feather cylinder 
(illustrated by lantern slides) shows the relation- 
ships of the growing parts of the feather, all of 
which arise from a thick ring of embryonic cells, 
the collar, the central opening of which (umbili- 
cus ) is occupied by the papilla. 

When the feather is plucked the papilla remains 
behind in the bottom of the follicle, consisting of 
a massive mesodermal core and a thin covering 
of a single layer of ectodermal cells from which 
the new feather is to be formed. 

///. Operations on the Papilla 

The purpose of this lecture is not a general re- 
view, but to present an account of experiments in 
which old methods of experimental embryolog)' 
have been applied for the first time to the develop- 
ment of feathers. 

The method of operating is to open the follicle 
so as to expose the papilla, and operate on the 
papilla with the finest iridectomy scissors. The 
follicle heals promptly. 

There are various kinds of operations that can 
be performed : ( 1 ) transplantation experiments, 
in which the papilla is completely removed from 



The Collecting Net was entered as second-class matter July 11, 1935, at the Post Office at Woods Hole, Mass., 
under the Act of March 3, 1879, and was re-entered on July 23, 1938. It is devoted to the scientific work at 
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28 



THE COLLECTING NET 



[ Vol. XVI, No. 139 



the follicle and transferred to some other site on 
the host; (2) defect experiments, in which parts 
of the papilla are removed, leaving the remainder 
within the follicle to develop; (3) isolation ex- 
periments, in which the papilla is divided longi- 
tudinally in determinate planes, and both parts 
left to develop within the same follicle; and (4) 
recombination experiments, in which part of the 
papilla from one follicle is removed and replaced 
with a supplementary part removed from another 
papilla. 

1. Transplantation Experiments 

The follicle from which a papilla has been re- 
moved does not regenerate a new papilla or 
feather. The papilla is the indispensable requisite 
for the formation of a feather. The feather papil- 
lae are tract-specific : if one takes a saddle papilla 
and implants it within a lireast follicle from which 
its own i:>a]3illa has Ijeen removed, the saddle 
papilla will produce a saddle feather in the breast. 
These feather germs or papillae behave just as 
specifically as different kinds of eggs. Wherever 
papillae are transplanted within the organism, 
there they produce the kind of feather peculiar to 
the site of origin. 

The papilla has a definite bilateral organization. 
If one dissect a jmpilla entirely free from all its 
attachments in the follicle, and rotate it 180° 
within the follicle, an upside-down feather results, 
i.e., ventral surface up and dorsal surface down. 
The papilla has no land marks on it like the 
frog's egg and so the operations must be made 
with reference to the position of the papilla with- 
in the follicle. It lies obliquely inclined with dor- 
sal surface up, \entral surface down. If one re- 
moves the ventral half by frontal bisection, the 
dorsal half will form a normal feather ; and if one 
divides the feather longitudinally and sagittally, 
a normal feather will also develop from a lateral 
half. Dorsal and lateral halves are thus complete- 
ly regulable. 

The situation is entirely different in the case of 
ventral halves. When the dorsal half of the pa- 
pilla is removed the remaining ventral half pro- 
duces one of two kinds of feathers, i.e., either 
"tuft" feathers in which there is no rhachis and 
the feather consists of a series of independent 
barbs, or "half-vane" feathers in which one lateral 
half is formed in a fairly normal fashion and the 
other lateral half is similar to tuft feathers except 
that the liarbs adjoining the half- rhachis are at- 
tached by their apexes and the bases are free. The 
production of the tuft feather is explained by the 
complete removal of the dorsal half of the papilla 
which is necessary for formation of a rhachis ; 
and the half-vane feather is explained by slight 
deviations right and left that leave narrow mar- 
gins of the dorsal half of the papilla, each induc- 
ing a half-rhachis. Correspondingly, it is found 



that about half of the half-vane feathers are right- 
handed and half left-handed. 

There is a clear analogy with the amphibian 
egg in respect to the behavior of dorsal and ven- 
tral halves respectively. 

Feathers with defects confined to the apex may 
be produced by amputation of the apical half of 
the papilla. These are to be interpreted as pro- 
duced not by loss of a prospectively limited seg- 
ment of the papilla, but by the delay in formation 
of the rhachis caused by the experiment, which 
produces in the apex of the feather conditions 
similar to the tuft and half-vane feathers. 

2. Isolation Experiments 

Twin feathers may be produced by dividing the 
papilla sagittally by a single cut and leaving the 
lateral halves in place. If each half develops in- 
dependently two normal and complete feathers 
are produced in a single follicle. But if the halves 
fuse together to a greater or less extent, as they 
usually do, twinning is confined to the apex of the 
feather. Successive generations of feathers from 
the same follicle produce types of separate or con- 
joint twin feathers similar to the first generation. 

3. Recombination Experiments 

Interesting chiiucra feathers may be produced 
by combining right and left lateral halves of pa- 
pillae within the follicle of one of them. This was 
illustrated specifically by chimerae between breast 
and saddle feathers in which one half-vane was 
saddle type and the other half-vane breast type. 

IV. Discussion 

In all of the experiments described above the 
defective papillae continue to produce the same 
types of defective feathers in successive genera- 
tions. In some cases as many as five successive 
generations have been recorded. There is a very 
pronounced lack of regulative ability on the part 
of operated papillae. 

From the experiments it follows that the mas- 
sive mesodermal core of the papilla acts as in- 
ductor on the ectoderm and determines the estab- 
lishment of a dorsal field in the ectoderm, within 
which the rhachis develops. Without this the de- 
velopment is always abnormal. 

From various lines of evidence it was concluded 
that the period of induction by the mesoderm is 
brief, and that thereafter the ectoderm constitutes 
a self-regulating system. The effects of the oper- 
ations within the definitive feather are due to dis- 
turbances of the inductive action on the ectoderm. 
This inductive action is general, not specific or 
mosaic. 

Out of hundreds of abnormal feathers produced 
experimentally no two are alike. However, all 
individual feathers produced in the defect experi- 
ments fall into the three main classes, "tuft" 



July 5, 1941 ] 



THE COLLECTING NET 



29 



feathers, "half-vane" feathers, and feathers with 
apical defects only. These are characterized by 
defects of the rhachis. 

Three kinds of barb defects occur in all three 
classes: branched, united and bundled. In the 
half-vane feathers, in addition to these l)arb ab- 
normalities, there occur also on the defective side 
of the half-rhachis reversed barbs which are at- 
tached by their apexes with their bases free. 

Branched barbs are due to fusion of the grow- 
ing barbs to form a single base which continues 
to grow as one, owing to disturbance of the regu- 
lar tangential movement. United barbs, i.e., with 
single apex and two or more bases, are due to 
division of the growth center of the base, each 
continuing to grow independently. If the fusion 
of the growth center that produces branching is 
followed after a time by division of the common 
base, "bundling" results. Reversed barbs occur 
only in connection with a half-vane and are due to 
the formation of a new ape.x adjacent to the half- 
rhachis and fusion of this apex with the out-grow- 
ing rhachis. 

From all this it follows that the rhachis is re- 
sponsible for the normal organization of the 
feather. It attracts barbs from both sides and is 
thus responsible for the formation of the ventral 
triangle to which formation of new barbs is con- 
fined in the normal development. It induces the 
formation of petioles on barbs that become at- 



tached to it and thus terminates the growth of the 
Ijarbs. It carries the completed barbs apically by 
its own growth and thus in general preserves the 
sequential order of barbs. It also determines the 
distinction between distal and proximal barbules ; 
and finally, it is to be noticed that each lateral 
half of the rhachis operates independently, as is 
shown by the half-vane feathers. 

The analysis of morphogenesis of the feather is 
then briefly as follows : 

1. The dorsal half of the papilla alone has the 
capacity of inducing the dorsal field in the ecto- 
derm, and this is the main morphogenetic func- 
tion of the papilla. 

2. The ectoderm of the papilla is capable of 
forming barbs which grow independently ; but the 
mesodermal induction is necessary for the deter- 
mination of the rhachis which controls the normal 
behavior of the barbs. 

3. The assembly of barbs on the rhachis oc- 
curs after their independent difi^erentiation. The 
abnormalities produced are in accordance with the 
principles of the special mechanism of develop- 
ment. 

Reference: Lillie, F. R., and Wang, Hsi. 1941. 
Physiology of development of the feather. V. Ex- 
perimental morphogenesis. Physiol. Zool., 14:103- 
134. 

(This article is an abstract of a lecture, illustrated 
with lantern slides, presented at the Marine Biologi- 
cal Laboratory on June 27.) 



INFORMAL NOTES ON EULIMA EMBRYOLOGY BY THE EMERITUS CURATOR 
OF THE M. B. L. MUSEUM 

George M. Gray 

(Continued from last issue) 



On August 10th the Tliyoiie having gone bad 
and no Eiilima eggs that I could discover, I threw 
out the Thyone, gave the Eiiliiua a clean bill of 
fare (sea water) and left them. The dish was 
covered with a glass plate, and they were alone. 

This morning, on looking at these same Eu- 
liina I discovered three fine-looking bunches of 
Eulima eggs. These had been laid between the 
time I changed them and this A. M., probably 
during the night. So far as I can prove this is 
the first real egg-laying of this particular lot of 
Eulima since they were brought to me on July 
31st and August 2nd. This is the second time 
I have had Eulima lay after being on Thyone, 
though I think that not all the Eulima went on 
the Thyone, but it would be interesting to see if 
any more are laid by these same snails. 

At 5 :30 P. M. I found two more egg capsules 
in the dish. The last two capsules laid since 
morning. Now five in all. 



August 12. About 8 o'clock A, M. this morn- 
ing there were four more egg capsules of Eulima 
in this same dish. This makes nine Eulima egg 
capsules in this dish, up to this morning. Still 
keep a glass over them. There are nine Eulima 
in the dish. About all the eggs are laid on the 
sides of the dish, the majority well up. 

August 13 — 9:15 A. M. Thirteen egg cap- 
sules. Some are in veliger stage. These thirteen 
egg capsules are from the nine Eulima last col- 
lected (July 31st and August 2nd). 

August 14 — 8 A. M. Two additional egg 
capsules were in the dish of the Eulima. This 
makes fifteen altogether. A few of the older ones 
of this lot are in the veliger stage. 

August 16. No eggs from the nine Eulima. 

September 1. No real good typical egg cap- 
sules from any Eulima on hand since last record- 
ed date. On August 31st I put a lot of the old 
Eulima in one finger-bowl and put a Thyone in 



30 



THE COLLECTING NET 



[ Vol. XVI, No. 139 



with them, to see if the change would be any en- 
couragement to egg-laying. A number of the old- 
est Eulima have died before now. 

About August 18th, three new freshly collected 
Eulima were brought to me. They were in a four- 
ounce jar. I left them in the jar and changed or 
partly changed the water, but no eggs up to Au- 
gust 31st. That day I changed the water more 
thoroughly, leaving them in the same jar. 

On September 1st about 8 A. M. I found one 
fresh laid egg capsule probably laid late yesterday 
or early this A. M. 

September 2. No more eggs from the three 
Etiliuia collected the 18th of August. Yesterday 
four fresh collected Eulima were brought in, but 
I did not get them until this A. M. Put them in 
a vessel of sea water and will hope for eggs. No 
eggs from Eulima which I put in with a Thyone 
yesterday. Everything else status quo. 

September 3 — about 9:20 A. M. The egg 
capsule from the Eulima of September 1st had a 
few separated individual veligers slowly moving 
about. No fresh eggs. 

September 4 — 8:30 A. M. More and faster 
moving veligers separately on the go this A. M. 
No new eggs and no eggs from the Thyone 
Eulima. Took Thyone out, added new sea water 
and await results. No eggs from any others. 

September 5. The veligers in the single egg 
capsule of the three Eulima of August 18th were 
moving around to some extent within the jelly 
mass, this A. M. At 5 :00 tonight they were much 
livelier but none have come through the surround- 
ing jelly walls. 

September 6. About same conditions prevail 
with Eulima as above. 

September 7 — 9:00 A. M. Veligers moving 
more rapidly, but all are within the surrounding 
walls of the egg capsule. All other Eulima are 
same as yesterday. 

September 16. Eulima veligers came through 
the walls this A. M. This means that it took from 
Sept. 5 to Sept. 16 for the larval Eulima to de- 
velop enough to come out of the capsule and swim 
around in the open sea. Some Eulima which I 
gave Dr. Alice Russell laid some eggs on Thyone. 
This I did not observe at all, though they might 
do so in their natural outdoor haunts. The Eu- 
lima seemed to get more pep and act better after 
a little sojourn on the Thyone. 

So far August has been the month which I 
have found to be ideal for egg laying of Eulima. 
though they may breed in July, but have had no 
time to investigate that early, though I hope to 
do this next year. 

Additional Observations 

February 28, 1941. Since late last Fall I have 
been carrying something like 12 to 15 Eulima 
oleacea alive to date in a finger-bowl. I changed 



the water nearly every day I was at the Labora- 
tory. There were times when they did not get a 
change of new water for 4 or 5 days, owing to a 
cold which kept me home. Also there were Sun- 
days when I did not go to the Laboratory and 
some few other times, but most of the time they 
had a daily change. I usually kept the bowl nearly 
covered with a glass plate. 

I think I have kept with them most of the time 
a live sea cucumber (Thyone briareus) of which 
they seem to be quite fond. I think I am now 
on the third one of these for the winter. The 
Eulima. when a Thyone is placed in a vessel with 
them, will begin to gather on it and in a few hours 
all or nearly all the snails will have attached 
themselves to the Thyone. on which they are par- 
tially parasitical. In changing the water I simply 
carefully poured off the water and either let new 
sea water run in until the dish is full or dip the 
bowl under water and thus fill it, sometimes mak- 
ing a double filling. In this way no true clean- 
ing has been done on the inside of the bowl. 
Whatever was loose in the bowl went out when 
the water was poured off. 

This morning I wondered if perchance there 
could be any eggs laid during their stay in the 
Laboratory during the winter. Much to my sur- 
prise I found something like a dozen egg cap- 
sules, some several days old and some only a day 
or two. 

March 1. I took out the egg capsules, placing 
them in another and smaller glass dish. In this 
way I could tell whether any new eggs were laid 
in the original finger bowl, and also could tell 
when the larval stages came through the walls of 
the egg capsule out into the water and really start 
on their individual careers. 

In the younger of these capsules the individuals 
lay closely packed together within the jelly-like 
walls of the capsule. As they grow older they 
begin to separate and start swimming about, but 
inside the walls of the capsule which seems to en- 
large. It takes several days before they are ready 
to come out. 

This morning I found three egg capsules laid 
near the surface of the water on the side of the 
bowl, so close that they seemed to touch one an- 
other. A second mass was on the bottom of the 
bowl, but I was not sure but what I had over- 
looked it in the previous lot. 

On March 3rd or 4th I found five newly laid 
egg capsules and on March 5th six more of them, 
laid during the night. Nearly all, I believe, are 
laid at night. 

March 6. Four more egg capsules were found 
on the sides of the finger-bowl this morning. 
This makes fifteen fresh capsules since the 3rd. 
Those of February 28 are still going. 



July 5. 1941 ] 



THE COLLECTING NET 



31 



Marcli S. Looked carefuHy tliis inoniinEj and 
decided two more egg capsules had been laid since 
the sixth. There are just 16 Enliuia in the bowl 
with the eggs, proving that one snail at least 
must have laid twfice. The veligers in the lot of 
February 28 are still swimming well, but all in- 
side the jelly. A few of the older ones have no 
movement and seem dead ; possibly the water got 
too warm. I don't think these snails lay eggs in 
winter in their natural habitat. I take it that the 
exceptionally warm temperature of the room 
started them laying, though of course it is possible 
that Eitlima might lay at this season as some of 
the Nudibranchs do. But I have my doubts about 
Eld una doing so. 

March 10. This morning the eg^ capsules had 
increased to 26 at least. This means that at least 
nine have been laid since the 8th. 

The whole number are in the finger-bowl with 
the Etiliina and the Thyone. I cannot say that 
any egg capsules have strictly been laid on Thy- 
one, but nearly all have so far been laid on the 
sides of the bowl, a few on the bottom. 

March 13. I took all of the Euliina and the 
Thyone from the finger-bowl this P. M. and put 
them all in another finger-bowl of clean sea water, 
leaving the egg capsules in the bowl in which they 
were laid. 

I have three egg capsules with veligers alive in 
them. These were of the lot laid in early March. 
Somehow they get about a certain age and die off 
without leaving the egg capsule. The water gets 
quite warm during the night and the water I use 
in changing is cold. First hot and then cold may 
have a pernicious effect, though at first they liven 
up when cold fresh sea water is given them. 
Squeezed some veligers from a capsule this P. M. 
They seemed to want to burrow in the bottom of 
the bowl. It must be that in their outdoor habi- 
tat they would naturally burrow in the sand or 
mud. 

Since putting the Eulima and Thyone in a clean 
bowl they have laid nine capsules of eggs. (This 
is up to March 24.) Four more laid two or three 
days after the change, and the others since. Have 
still a few of the veligers from the older lot living 
on March 24 (inside the capsule). 



jAfter this there seems (o be a blank for I did 
nothing further with the lot except to put them 
in one finger-bowl and set them in my salt water 
sink, and now and then letting a little salt water 
run on them. Thus time passed on until this 
A. M., June 23rd. On looking at the Eiilima to- 
day which I had put in a finger-bowl in the salt 
water sink some weeks ago I found eleven egg 
capsules in different stages, some quite young, 
some in the veliger stage. Evidently these Eu- 
lima had been in the sink since last March, and 
were not in running water. The only change they 
had was turning a small sea water hose into the 
bowl for a few seconds every other day or so, but 
I did not examine the snails or even look in the 
dish in all the time they were left in this condi- 
tion. So I was surprised to find the capsules. 
There were seven dead shells of Eitlima, and five 
live snails. I cleaned the dish, leaving the eggs 
and took out the dead snails' shells and put liack 
the live snails with the egg capsules in the clean 
sea water, and they are still there. 

This ends for a time the history of this little 
gastropod mollusc. There is much more to learn 
about its later larval development, how the shell 
is formed, etc. This we hope to learn later. 
* * * 

Since the above article went to press I have 
made the following observations. A collection of 
10 Euliina was brought to me on June 25 th. I 
put them in a clean finger-bowl of sea water. On 
the 26th there were no eggs laid. 

On June 27th there were 8 egg capsules. 

On June 28th there were 10 egg capsules. 

On June 29th there were 21 egg capsules. 

On June 30th there were 39 egg capsules. 

On July 1st there were 52 egg capsules and 
one dead Eulima. I think the eggs were laid by 
the 9 as the dead one had probably been dead 
when brought in with the other nine. On July 
2nd there were 86 egg capsules with the nine liv- 
ing Eulima. 

This extends the Ijreeding range for practically 
another month and perhaps Eulima may even 
breed in May. This will have to be ascertained 
another year. There is conclusive proof that 
each snail lays several egg capsules, for here are 
86 egg capsules, and only nine snails for the job. 



PROTOPLASMIC STREAMING 

(Continued from page 25) 



tarded to an equal extent and are finally abolished 
as the pressure is gradually raised to about 5,000 
lbs. per square inch. 

The least common denominator in these ex- 
periments appears to be that pressure has a 
liquefying action upon gelated parts of protoplasm 



generally. Or, to state things conversely, — high 
pressure inhibits gelation processes which normal- 
ly must occur in cells when the streaming of the 
more fluid protoplasm is being motivated. In 
any event, it has been found that the extent to 
which the streaming is inhibited corresponds ex- 



Z2 



THE COLLECTING NET 



[ Vol. XVI, No. 139 



actly to quantitative measurements of the curtail- 
ment of the gelation processes. 

An interesting assemblage of apparatus was 
necessary for the experiments. The pressure 
pump was constructed from a hydraulic jack of 
the type ordinarily used for lifting of heavy 
trucks. The thick-walled pressure chamber was 
made of stainless steel and provided with heavy 
glass windows. Since an ordinary microscopic 
oljjective would not he suitable for viewing tis- 
sues through so thick an intervening layer of 
glass, a special objective, with a working distance 



of more than half an inch, was necessary. This 
objective, used with an inverted type of micro- 
scope, gave good images at a magnification of 600 
diameters. And lastly, the measurements oh pro- 
toplasmic fluidity required a special centrifuge- 
pressure chamber. A suitable valve permitted no 
loss of pressure, even after the chamber, having 
received its charge from the pump, was discon- 
nected and placed in the centrifuging equipment. 

(Summary of paper presented at "The Symposium 
of Protoplasmic Structui-e" at the meetings of the 
American Association for the Advancement of 
-Science, December 30, 1940.) 



GRAFTING OF LIMBS IN PLACE OF THE EYE IN AMBLYSTOMA 

Dr. Jean Piatt 

Departments of Anatomy, University of Vermont, Burlington, Vermont and College of Physicians 
and Surgeons, Columbia University, NeT(.< York City 



When a forelimb rudiment is grafted to the 
head region of salamander emlDryos, the resultant 
Hmlj is usually well developed and capable of 
movement. The range and vigor of movement in 
the transplant vary considerably from case to 
case, dependent upon the specific anatomical rela- 
tions existing between graft and host and upon 
the particular region of the head involved. A 
correlation of movement in the transplant with 
that of other muscle groups usually occurs, but 
the reason for this phenomenon is not entirely 
clear. Particularly is this the case when limbs 
have lieen transplanted in place of an eye. 

Nicholas ('29, '30, '33) made homoplastic and 
heteroplastic forelimb grafts to the eye region in 
Amhlystouta. In general, the movements of such 
limbs were found to be rather poor but more dis- 
tinct in some cases. Some of the transplants ex- 
hiliited individuated movements of the various 
])arts. Nicholas states that when movement of a 
limb occurred, it was always in coordination with 
ocular movements of the contralateral eye. 

In order to prevent the normal appearance or 
regeneration of the eye muscles, Nicholas cleaned 
the wound of all mesenchyme cells clown to the 
wall of the brain. .Although he does not actually 
state that the eye muscles were eliminated in the 
specific animals in question, it is assumed from 
a supplementary e.xperiment that such was the 
case. The transplant in a number of cases was 
innervated partly by eye muscle nerves, chiefly 
the oculomotor. Nicholas himself states that in 
the event eye muscles should regenerate and in- 
sert on the base of the limb, the experiment would 
be vitiated, since the eye muscles themselves, not 
the eye muscle nerves, would be causing the co- 
ordinate movements. Nicholas interprets his re- 
sults as an argument against Weiss' Resonance 
Theory, or theory of Homologous Response. 

Weiss ('36) in an analysis of Nicholas' work 



makes the suggestion that in those cases which 
demonstrated movement synchronous with the eye 
some of the eye muscles might still be present and 
insert on the limb, thus accounting for the co- 
ordinate movements observed by Nicholas. In 
order to test this Weiss transplanted larval limbs 
in place of the eye and found that when the al- 
ready developed eye muscles were not cut away 
in the operation they found new insertions on the 
transplant. The movement of these limbs was 
extremely weak and stereotyped, and the limb 
moved only as a whole. Weiss concluded that 
such an arrangement of the eye muscles might 
]M)Ssibly explain the seeming discrepancy between 
his theory of Homologous Response and that of 
central coordination postulated by Nicholas. 

It has been the purpose of this investigation to 
attack this problem anew, repeating the experi- 
ments of Nicholas but in a slightly altered form 
and to make a careful examination of both limb 
movement and the anatomy of the region in- 
volved. 

Forelimb rudiments were grafted in place of 
the eye in embryos of Amblystoma punctatum. In 
series LHR-1 only the eye and covering ectoderm 
were removed ; in series LHR-2 the same opera- 
tion was performed but in addition the wound 
was cleaned of all mesenchyme cells down to the 
brain wall. Observations were made on the 
amount and type of limb movement and the full 
grown lar\-ae were then sectioned and a study 
was made of the muscles and nerves in the region 
of the transplant. 

No significant difference between the two series 
was revealed, either in movement or anatomy of 
the region involved. Eye muscle tissue was found 
to be present in every case studied, and individual 
muscles could often be identified. The eye muscles 
either inserted directly on the cartilage of the 
transplant or in its immediate vicinity. It is 



July S, 1941 ] 



THE COLLECTING NET 



33 



thought that contraction of these muscles would 
account for the feeble, stereotyped movements oh- 
ser^'ed in the grafted limbs and for the fact that 
the limb movements were nearly always corre- 
lated with movements in the opposite, intact eye. 
Limbs were always innervated by the opthalmic 
Ijranches of the V-VII complex, and never by the 
eye muscle nerves directly, if at all. 

My experiments sustantiate the contention first 
expressed by Weiss, that when limbs are put in 
place of an eye, their movement is extremely weak 
and stereotyped ; further, that it is diiTicult to, re- 
move all the eye muscles in such an operation and 



that the coordinate movement of the hmb with 
the opposite eye is caused by the remaining eye 
muscles gaining an insertion onto the liase of the 
trans]ilant. If this is so, then the coordination of 
limlj with eye in such cases does not mean that 
non-homologous muscles are functioning in a 
homologous response because of innervation by 
nerves of an identical nature. 

(This article is a summary prepared for the press 
of a paper presented before the American Society 
of Zoologists at the meeting of the American Asso- 
ciation for the Advancement of Science on December 
30, 1940.) 



DEPARTMENT OF CHEMICAL SUPPLIES AND SCIENTIFIC APPARATUS 



Standard Solutions: 

The Chemical Room has charge of the order- 
ing, storing, and distribution of ordinary glass- 
ware, simple laboratory apparatus, reagents, and 
chemicals. Drugs, dyes, and certain rarer com- 
pounds owned by the Laboratory, are all kept in 
the Chemical Room, ?^8, Brick Laboratory Base- 
ment. These may be used by investigators and 
classes during their work at the Laboratory. 

Supplies to be used elsewhere than at this Lab- 
oratory are not furnished by the Chemical Room. 
All such, e.g., bottles, jars, slides, covers, labels, 
drawing materials, and instruments — can usually 
be purchased from the Supply Department. 

The Chemical Room Staff makes up various 
histological and photographic solutions, and cer- 
tain standardized reagents used by investigators 
and classes for ordinary biological and chemical 
work. Such solutions and reagents are described 
in "Formulae and Methods," III, 1936. The 
Chemical Room does not undertake special chem- 
ical work for individuals or classes nor the prep- 
aration of solutions, requiring unusual time or 
facilities. However, for the period from June 24 
to September 1, the chemist will undertake to 
standardize a limited number of special solutions 
for individual investigators ; and to determine the 
pH of a limited number of aqueous solutions. It 
is preferable to submit at least 10 ml. quantities 
of well-buffered solutions or 20 ml. quantities of 
unbuffered solutions, although determinations will 
be made on less solution, if necessary. The sam- 
ples should be placed in clean, stoppered contain- 
ers which are labeled with the name of the investi- 
gator, the room number, the approximate pH if 
known, and a description of the contents, if of a 
hazardous nature — cyanide, for instance. The 
samples should be left at the Chemical Room be- 
fore noon. The results may be obtained at the 



Chemical Room the following day. The number 
of determinations for any one investigator is 
necessarily restricted. Investigators expecting to 
use such solutions or standardized reagents after 
September 1 are requested to notify the Chemical 
Room, if possible, before August 24. 

Special Supplies : 

Dry ice may be obtained at cost in limited 
amounts through the Chemical Rooni. For dry 
ice, rare chemicals, etc., ample advance notice of 
at least three days is required. In the case of ex- 
pensive reagents such as osmic acid, gold chlor- 
ide, platinum chloride, special organic compounds 
and dyes or stains the Laboratory makes a charge 
at current prices for all above a reasonable 
amount used by an investigator. This rule may 
apply also to any other materials which are 
scarce, difficult to obtain, not likely to be required 
by other investigators, or are requested in un- 
usual quantities. 

Draughting supplies, portable microscope ac- 
cessories, dissecting equipment, surgical instru- 
ments including syringes, needles, etc., are not 
available for loan but may be purchased through 
the Supply Department. Easily transported ap- 
paratus such as stop-watches, cameras, and de- 
vices of special application should be provided by 
each investigator (before departure for Woods 
Hole) unless he is assured that they can be fur- 
nished by the Laboratory. 

Window Service, 1941 : 

June 30 to August 30 — 8 :30-12 :00 a.m./l :30- 
4:30 p.m. Saturdays— 8 :30 to 12:00 a.m. 

Holidays excepted. 

September 2 to 27 — 11:00-12:00 a.m./3 :30- 
4 :30 p.m. 



34 



THE COLLECTING NET 



[ Vol. XVI, No. 139 



The Collecting Net 

A weekly publication devoted to the scientific work 
at marine biological laboratories. 

Edited by Ware Cattail with the assistance of 
Boris I. Gorokhoff and Judy Woodring. 

Entered as second-class matter, July 11, 1935, at 
the U. S. Post Office at Woods Hole, Massachusetts, 
under the Act of March 3, 1879, and re-entered, 
July 23, 1938. 



SYMPOSIUM ON DEVELOPMENT AND GROWTH 

The following is the program of the Sympos- 
ium of the Society for the Study of Development 
and Growth to be held at Dartmouth College, 
Hanover, N. H., from July 7 to 11. 

General Topic: Patterns. 
Monday, July 7th 

Morning Session: Francis O. Schmitt (Washing- 
ton University, St. Louis.) "Patterns of Protein 
Molecules." Configurations, orientations, and prop- 
erties of protein and conjagated protein components 
in the ultrastructure of cells and tissues. Supple- 
mentary data from mono- and multifilms and crys- 
talline protein. 

Afternoon session: Vance Tartar (University of 
Vermont). "Intracellular Patterns." Facts and prin- 
ciples concerning patterns exhibited in the morpho- 
genesis and regeneration of ciliate protozoa. 

Tuesday, July 8th 

Morning session: Kenneth B. Raper (U. S. Dept. 
of Agriculture). "Patterns of Primitive Cell Col- 
lectives." Developmental patterns in simple slime 
moulds. 

Afternoon session: A. F. Blakeslee (Carnegie In- 
stitution, Cold Spring Harbor). "Growth Patterns 
in Plants." 

Wednesday, July 9th 

Morning session: N. J. Berrill (McGill Univer- 
sity). "Growth patterns in Lower Animals." Spat- 
ial and temporal patterns in colonial organisms. 

No session in the afternoon. 

Thursday, July 10th 

Morning session: A. H. Hersh (Western Reserve 
University). "Heterogenic Growth." The ontogene- 
tic and phylogenetic significance of diff'erential rates 
of growth. 

Afternoon session: L. C. Dunn (Columbia Uni- 
versity). "Abnormal Growth Patterns, vdth Special 
Reference to Genetically Determined Deviations in 
Early Development." 

Friday, July 11th 

Morning session: Paul Weiss (University of Chi- 
cago). "Nerve Patterns." Analysis of the factors 
controlling the development of the nervous system 
as an example of complex pattern formation. 

Afternoon session: Assignment not yet decided. 



THE CHILDREN'S SCIENCE SCHOOL 

The Children's Science School opened Monday, 
with a registration of 57 children. More have 
been registered during the week. The registration 
was followed by a showing of lantern slides by 
Mr. Lower, which was greatly enjoyed by the 
children. 

The teachers this summer are Miss Helen 
Smith, who is also director, Mr. Reginald Mac- 
Haffie, and Mr. and Mrs. George G. Lower, all 
of whom were here last year. 

At the opening meeting Monday afternoon, the 
teaching staff gave most interesting outlines of the 
courses they are giving this year. In Miss Smith's 
marine ecology course, the specimens are collected 
on frequent field trips and identified and pre- 
served at the school. The biology course is also 
presented by Miss Smith. It consists of an intro- 
duction to the structures and functions of ani- 
mals, and to some of the more important biologi- 
cal principles. Lectures are supplemented by ex- 
periments and dissections of interesting forms. 

Mr. ^lacHaffie is hoping to do quite a bit of 
culturing of insects in his entomology course, with 
ant colonies, bees. etc. For the junior laboratory, 
he has several problems to work out, on which 
little research has been done. Genetics and em- 
bryology are particularly emphasized. 

Mr. Lower is to have his very original system 
of charts, with teams, tests in various subjects, 
and awards with special credit given for collec- 
tions. These are for both the Water Life and 
Nature Sttidy classes, and he is also thinking of 
adding a bit on weather, tides and navigation to 
the more advanced class, explaining the buoys, 
fog signals, etc. 

Mrs. Lower in her Introduction to Nature 
Study initiates the beginners in observation, col- 
lecting, classification, etc., and says that, with a 
little training, the youngest children become keen 
observers and make very creditable collections. 
She plans to instruct them early in the class on 
aquaria and keeping sea life at home. 

These previews of the courses make it appear 
that the Science School has a particularly stimu- 
latingf season ahead. — Mrs. S. McMnrtrie. 



CURRENTS IN THE HOLE 

At the following hours (Daylight Saving 
Time) the current in the Hole turns to run 
from Buzzards Bay to Vineyard Sound: 
Date A. M. P. M. 

July 5 12:59 1:16 

July 6 1:56 2:12 

July 7 2:54 3:08 

Tuly 8 3:48 4:00 

"julv 9 4:37 4:53 

"July 10 5:28 5:40 

July 11 6:13 6:30 



July 5, 1941 ] 



THE COLLECTING NET 



35 



ITEMS OF INTEREST 



Dr. Eugene F. DuBois, professor of medicine 
at the Cornell LTniversity College of Medicine, has 
heen appointed professor of physiology and head 
of the department of biochemistry and physiology 
at the College. He succeeds Professor Detlev W. 
Bronk, who returns to his post as director of the 
Johnson Foundation for Medical Physics at the 
University of Pennsylvania. 

Dr. Donald P. Costello, assistant professor 
of zoology at the University of North Carolina, 
will be on leave of absence next year. He has 
been appointed a Rockefeller Foundation fellow 
and will work at the School of Biology at Stan- 
ford University. 

The Carnegie Corporation of New York has 
granted to Dr. L. J. Milne, associate professor of 
biology at Randolph-Macon Woman's College, 
about $900 with which to obtain special equip- 
ment for biophotography. This summer at Woods 
Hole he is collaborating with Dr. C. L. Parmen- 
ter in presenting cell division of salamander epi- 
dermis as animated diagrams on motion picture 
film. 

Dr. T. K. Ruebush is leaving for Washington, 
D. C, where he has been called to active duty in. 
the Medical Corps of the U. S. Navy. 

Among the persons who attended the Sympos- 
ium on Genes and Chromosomes at the Biological 
Laboratory at Cold Spring Harbor and who will 
spend the summer at Woods Hole are : Dr. and 
Mrs. P. W. Whiting, Dr. C. W. Metz, Dr. Har- 
old H. Plough, Dr. R. Ruggles Gates and Dr. 
Morris Harnly. 

Notes from the Bureau of Fisheries 

Mr. Robert A. Goffin and his stafif had 
planted 1,293,000 mackerel fry at the end of last 
week. The mackerel were stripped at the traps 
and the eggs fertilized and brought to the Hatch- 
ery. The Bureau of Fisheries carries out this 
conservation measure each season. 

A new attraction at the Fisheries pool is the 
blue shark, approximately six feet long, which 
was taken in a fish trap in Buzzards Bay off Pen- 
zance Point. It is unusual for this shark to come 
so far inland. In addition, there are two seals 
and a few dogfish in the pool. 

The registration for the month of June at the 
Bureau of Fi,sheries was 3,401. From Julv 1, 
1940 to June 30, 1941, 25,099 persons wrote their 
names in the visitors' book. In general, Mr. 
Goffin finds that about one in four persons who 
come to the aquarium register there. 



y\t the Durham meeting of the American Asso- 
ciation for the Advancement of Science, the pro- 
gram of the Section on Medical Sciences included 
a paper on properties of skeletal muscle fibers by 
Dr. F. J. M. Sichel, assistant professor of physi- 
ology at the University of Vermont College of 
Medicine. Mrs. Sichel went to New Hampshire 
to read the paper for him owing to his conflicting 
engagement with the physiology course. 

Dr. Albert Navez, of the Milton Academy, 
presented a paper on "Biology, the Science of 
Life (Plea for the Experimental Method)," on 
June 25 before the National Association of Biol- 
ogy Teachers at the Durham meetings. 

The Spencer Lens Company is holding its ex- 
hibit at the Canteen Building until Friday, June 
11. 

The program for the weekly phonograph rec- 
ord concert at the M.B.L. Club next Monday is 
as follows : Beethoven, "Overture to Leonore, No. 
3"; Beethoven, "Symphony No. 5"; intermission; 
Brahms, "Symphony No. 8." 

Dr. T. K. Ruebush, the secretary-treasurer of 
the Tennis Club, announces that a tennis tourna- 
ment will be scheduled this summer if enough 
people show interest and apply to take part in it. 
Anyone who 'wishes to enter such a tournament 
should see Dr. Ruebush, Dr. D. E. Lancefield, or 
Albert Stunkard. 

The Woods Hole Choral Club opened its sea- 
son with a very successful rehearsal Tuesday 
night at the Canteen Building of the Bureau of 
Fisheries. A group of over thirty persons con- 
nected with the Laboratory was present, and 
practiced four of the songs which will be sung at 
the annual concert late in August. The next re- 
hearsal of the Club will be held on Tuesday. 

ADDITIONAL INVESTIGATORS 

Clark, Eleanor L. assoc. anat. Pennsylvania Med. Bi- 
ll?. 

Erlanger, Margaret instr. West Virginia. Br 312. 

Finkel, A. J. grad. asst. zool. Chicago. Br 322. 

Gelback, Elizabeth L. asst. proto. Yale. Br 323. 

Hamilton, H. L. res. asst. emb. Hopkins. Br 324. 

Harnly, M. H. assoc. prof. biol. New York. Br 344. 

Horn, Annabelle grad. asst. zool. Pittsburgh. Rock 7. 

Keefe, E. L. res. asst. zool. Washington (St. Louis). 
Br 217-j. 

Klotz, J. W. grad. zool. Pittsburgh. Rock 7. 

Michaelis, L. mem. Rockefeller Inst. (New York). 
Br 207. 

Mullins, L. J. asst. phys. Rochester. Br 322. 

Netsky, M. Pennsylvania Med. Br 205. 

Sandow, A. asst. prof. biol. New York. Br 344. 

Spratt, N. T., Jr. res. asst. emb. Hopkins. Br 324. 

Tracer, W. assoc. Rockefeller Inst. (Princeton). Br 
■^208. 

Weaver, Margaret A. grad. zool. Texas. Br 312. 



36 



THE COLLECTING NET 



[ Vol. XVI, No. 139 



PHYSIOLOGY 

Our second two weeks' period of work is now 
in full swing. The tables have been turned and 
now we are showing Dr. Fisher how to run the 
Warl)urg apparatus. It does our hearts good 
these days to see him taking down manometer 
readings, and cleaning vessels. We hope his re- 
sults are better than ours were. 

The home-run king. Dr. Parpart. began the lec- 
tures this week, talking on permeability, and the 
composition and structure of cell membranes. 
Thursday Dr. S. C. Brooks lectured to the class, 
the sul)ject being "Some Problems in Permeabil- 
ity." Our first guest lecturer was Dr. L. V. Heil- 
brunn, who spoke last Saturday on the theory of 
muscular contraction with special reference to the 
role of ions. It was he who quoted the great 
physiologist and chemist. Loeb, as saying that if 
you don't have brains, use apparatus. We use 
apparatus. 

There are now four sections running parallel 
under Drs. Parpart, Kempton and Ballentine. 
One group of Dr. Parpart's students is studying 
permeability using dogfish red cells and arbacia. 
Another group is determining the total lipid and 
cholesterol contents of dogfish ghosts. Here again 
is something we can't see. A third group is 
studying the hemolytic effects of detergents, and 
a fourth is using cleavage curves of arbacia eggs 
to determine permeability. 

Part of Dr. Kempton's class may be found in 
the liasement micro-manipulating, while the others 
are upstairs learning why the kidney. Dr. Bal- 
lentine's students are determining the intercellu- 
lar proteases during development of arliacia. and 
the distribution of dipeptidase in arbacia eggs. 
This is running along smoothly because at least 
one member of the group has worked on eggs be- 
fore, — hen's eggs. To quote this particular mem- 
ber, "Arbacia eggs are very tiny." 



CLASS NOTES 

In the blood gas studies group, with the aid of 
Drs. Parpart and Ballentine, oxygen dissociation 
curves of limulus hemocyanin, spectrum absorp- 
tion curves in oxygenated hemocyanin, and mano- 
metric methods are being emphasized. 

"Come on Physie, keep 'em busy!" yelled the 
sideline physiologists, as they cheered their team 
on to victory. It all happened Monday evening 
when we met our friends the embryologists out 
upon the baseball field. We are happy to say that 
we came out on top, with a score of 18 to 14, but 
not without a few anxious moments. It was a 
good game, from both sides, and it was only 
througli the able pitching of Bob Harrison, who 
also hit a home-run and brought three others in ; 
the whole-hearted cooperation of our class and the 
faculty representative, Dr. Kempton ; and the 
brains back of the team (those of manager Bill 
Keezer) that we accomplished the deed. 

— And then came Mr. Trinkaus up to bat for 
the embryologists. Strike one ! Strike two ! ! 
Strike three ! ! ! — Just another sad story of The 
Third Out, or With the Bases Full and (all due 
apologies to the poem of the same name hanging 
in Dr. Fisher's office) Nobody Ohm. 

The embryologists join us in thanking Ted for 
doing such a good job as umpire. 

We must not forget the game played last Fri- 
day afternoon — faculty et all. Maybe it was the 
practice they gave us that helped us win on Mon- 
day. The highlights of the game were Dr. Par- 
part's home run and Dr. Fisher's striking out. 

We are looking forward with a great deal of 
pleasure to our class picnic which has been 
planned for next Tuesday. Arrangements are 
lieing made by the committee. Bob Harrison and 
the Black Phantom (his car). — J. E. H. 



BOTANY CLASS NOTES 



Si)metimes I have to wonder why 
The arU.st's life I did deny, 
For plants, I find, are not elating 
And sliinly Chora's most nauseating. 

True as this is in lab, collecting is another 
story and a much more pleasant one: the Cutty- 
hunk expedition was a perfect vacation trip. A 
picturesque little fishing village clinging to the 
hillside, the stone-walled road snaking through it 
and up into nowwhere, the Coast Guard look-out 
box on the highest point, the flashing twin lakes 
(Chara gold mines), miniature orchids in the 
deep sphagnum gullies, a storm-worn monument 
to Gosnold (probably rich in aerial algae), the 
long strand of bright, white sand, intensely blue 
water, lazy gulls — al! enough to convince one of 
the authenticity of a National Geographic illus- 



tration. But romanticism went by the board 
when nineteen botanists plunged into the waist- 
deep muck of Euglena Pool and waded through 
the waters of Sheep Pond. That night in lab we 
learned that this disagreeable muck is an algolo- 
gist's paradise, having species brought from for- 
eign parts by none other than the lowly sheep 
that bathe there on arrival from the mainland. 

Class field trips, however, are not the only ex- 
ercise we get. "I'll never in all my life forget the 
sight of Monti standing on the dock with six girls 
around him, wildly gesticulating and almost hid- 
den by the fog!" reminisced Connie Stanton in 
recalling the Monti Expedition to Nonamesset 
Island. If you happened to see a gentleman strid- 
ing down the Main Street last Saturday after- 
noon, followed by six somewhat feminine-looking 



July 5, 1941 ] 



THE COLLECTING NET 



37 



creatures, it was not a sultan with his harem, but 
just an innocent lad leading a bevy of blue-jeancd 
botanists to the Oceanographic Pier. The former 
algology student had in some way procured two 
skiffs for the trip. Being a true celibate at heart, 
Monti allotted one to Bob Thorne and very gen- 
erously sent him out to sea with only two of the 
fair sex for crew, setting out with the remaining 
half dozen himself. The greatest fun occurred 
when the waves — choppy and very wind-blown — 
proved too much for the overloaded rowboat and 
forced the hero to sacrifice his adventurous plans 
for the delights of retaining his harem. Bob, 
meanwhile, faring better with his smaller group, 
was able to master the ocean, although the de- 
feated seven finally persuaded him to quit his ef- 
forts in favor of a hike to Nobska, via the more 
tranquil terra firma. 

Such ideas, especially those of hiking, are typi- 
cal of the Botany Class. As if the all-day work, 
five and a half times a week, were insufficient, 
certain fanatics in the course find it necessary to 
devote part of the Sabbath to quenching their 
thirst for knowledge. So that bright and early — 
about 8:30 A. M. — on beautiful foggy Sundays, 
Bob Thorne may be seen leading his devotees out 
to gather the fair posies of local habitats. Last 
Sunday, however, the presence of Charlie Abbot, 
welcomed by everyone, seemed to worry Mr. 



Thorne to such an extent that he retired within 
his anti-female shell and did a much more thor- 
ough job of collecting. 

A collection of algae specimens is, of course, 
the ultimate tangible result of this class, but 
equally interesting tangents are continually add- 
ing to the less mountable, general knowledge that 
the botanists are also collecting. An extra bit 
was added last Thursday night when Bob Wil- 
liams gave a very informative lecture about carbo- 
hydrate metabolism in the large brown algae. 
What with his illustrative graphs and his slightly 
, shaky knowledge of the larger details of organic 
chemistry, the audience found the seminar far 
from dull. Tea, as usual, climaxed the program. 

Evening teas, though, are hardly long-lived 
enough to stave off the starvation pangs in mid- 
afternoon. Several would-be-fortunate recipients 
of boxes from home — chiefly Elaine Katz — were 
completely submerged and left entirely bare of all 
food-stuffs when the hungry class descended upon 
them. Which all goes to prove something, one 
would suppose. At any rate, there's no doubt 
that the competitive spirit runs abnormally high 
at such times — particularly between Bob Muir 
and Sam Salvin who with all the grace of true 
gentlemen, inevitably consume the greatest amount 
of the booty. But then it's every man for him- 
self, with the fastest one the winner. — /. W. 



EMBRYOLOGY CLASS NOTES 



A new idea was brought to our attention Tues- 
day morning by Dr. Goodrich in his talk on the 
patterns of chromatophores in goldfish. Appar- 
ently one can have one's fish monogrammed ac- 
cording to order by transplantations of scales. 

The squirting squid was squarely squelched 
when Dr. Hamburger demonstrated "How Squids 
Squirt." Looking at the various sketches made 
by the class we were carried back to our child- 
hood stories and Walt Disney's interpretation of 
Mother Goose. To become more scientific, the 
development of the squid eye was of interest be- 
cause of its similarity to the mammalian eye, al- 
though its parts are not homologous. 

Dr. Hamburger's lecture on embryonic induc- 
tion where the history was traced and theories old 
and new presented was one of the high spots of 
the week. 

Coelenterates were presented by Dr. Ballard, 
whose humorous comments on the life histories 
of various members of that Phylum were enthu- 
siastically received. How nice to lead a double 
life as they do by metamorphosis ! 

Tunicates are our ancestors or so we were told. 
Apparently they are the other extreme of the 
]5hylum Chordata, to which Homo sapiens be- 
longs. 



The week ended with a lecture by Dr. Schotte 
(sans chapeau — he lost it) opening the work on 
echinoderms. 

The high spots of the unscientific week were 
initiated by our first baseball game with the crew. 
We really didn't do so bad considering their pre- 
vious experience. The embryologists work, the 
crew — well, maybe. 

The Second Trinkaus has arrived and was 
promptly put to work collecting towels which Ted 
distributes. While we are on the subject — the 
First Trinkaus, together with an eminent in- 
structor received a biological baptism early in the 
week when a canoe upset in front of the M.B.L., 
a very appropriate place. One watch, the only 
casualty, was given first aid — dehydration. 

The dance on Saturday night was quieter than 
that of the preceding week. Embryology was well 
represented. 

Our cultural life was taken care of by a con- 
cert at the M.B.L. Club which, added to the har- 
mony by Jack, Tom and Dr. Ballard in the lab at 
night, greatly increased our musical appreciation. 

As this goes to press we are feeling greatly 
encouraged. The Softball score between classic 
rivals is swinging in our favor, as was shown in 
the game tonight with the people in the "back 
room." — P. H. and E. K. 



38 



THE COLLECTING NET 



[ Vol. XVL No. 139 



The A. B. a of Woods Hole for 1941 

All Schedules Set to Daylight Saving Time — Bold Type Indicates P. M. 



POST OFFICE 

Week Days Sundays 

Mail Arrives 8:00,10:55,3:45,7:15 10:35 

Mail Closes 6:30,9:30,5:00 5:00 

Station Open 6:00 to 8:00 10:30 to 5:15 



Window Service 7:30 to 6:00 

All mails should be deposited at least ten 
minutes before closing time to insure dispatch; 
registered letters should be deposited fifteen 
minutes before closing time. 



TELEGRAPH OFFICE 

Weekdays 
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Mondays, Wednesdays and Saturdays 
3:00 to 5:00 
7:00 to 9:00 

RELIGIOUS SERVICES 

Church of the Messiah (Episcopal) - 

Sundays: 8:00 Holy Communion; 11:00 
Morning Prayer (Choral Eucharist, first 
Sunday in the month). 
Holy Days: 8:00, Holy Communion. 

Methodist Episcopal Church 

Morning Worship, 11:00. Church School, 
10:00. 

First Orthodox Congregational Church 

Evening Service, 7:30. 

St. Joseph's Roman Catholic Church 

Mass: Sundays, 6:45, 9:30. 
Weekdays, 7:00. 







TRAIN 


SCHEDULE 


* 






Wood-, Hole 
llostoii 


Weekdays 
7:0(; 
i) : 1 5 


Weekdays 
10:15 
12:35 


Ex. Sat. 
& Sun. 

12:55 
2:52 


Weekdays 
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Sundays 
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Sundays H 
7:55 
9:55 


Hohtoii 
Woods Hole 


Weekdays 

8 :20 

10: 15 


Sundays 

S:35 
10:15 


Saturdays H 
12:25 
2:30 


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1:05 
3:25 


Ex. Sat. 
& Sun. 
4:00 
5:59 


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7:10 


*A11 trains stop 


at Falmouth. 


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7:30 




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9:20 


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8:66 


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4:45 




Vinevard Haven 


6:10 








6:00 






Oak Blufis 




9:00 


4:00 


4:30 




6:45 


9:00 


Woods Hole 


6:55 


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5:00 


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* Schedule effective 


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incl. 




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tDoes not run Labor Day. 






SAlso runs Labor Day. 







July 5, 1941 ] 



THE COLLECTING NET 



39 




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THE COLLECTING NET 



[ Vol. XVL No. 139 





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July 5, 1941 ] 



THE COLLECTING NET 



41 



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42 



THE COLLECTING NET 



[ Vol. XVL No. 139 




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•44 



THE COLLECTING NET 



[ Vol. XVI, No. 139 




THROUGH the cold dank dusk a watcher 
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THE SOURCE OF PANCREATIC JUICE 
BICARBONATE 

Dr. Eric G. Ball 
Associate Professor of Biochemistry, 
Harvard Medical School 
Pancreatic juice that is rapidly secreted is in 
osmotic equilibrium with blood plasma but con- 
tains mainly sodium bicarbonate as its inorganic 
constituent. This means that 
pancreatic juice may contain 
five to six times the quantity 
of bicarbonate ion that is 
found in blood plasma. What 
then is the source of this juice 
bicarbonate ? Two possible 
sources exist. One is the bi- 
carbonate of the blood flowing 
through the gland. The other 
is the COo produced by the 
metabolic processes of the 
gland itself. At the start of 
this investigation we were in- 
clined to the view that a con- 
siderable part of the bicarbon- 
ate of the juice was derived 
from the metabolic CO2 of the 
gland itself, for two reasons. 
First, because the amount of 
bicarbonate in the juice varies 
with the rate of juice secretion. 
The more rapid the rate of secretion the higher 
the bicarbonate content. Such a relation can be 
interpreted to mean that {Continued on page 55) 



% 1. Calcnbar 



TUESDAY, July 15, 8:00 P. M. 
Seminar: Dr. L. B. Clark: "A 

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the Moon Influences Reproduc- 
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Worm." 

Dr. Paul S. Galtsoff: "Accumula- 
tion of Manganese and the Sex- 
ual Cycle in Ostrea virginica." 

Dr. R. M. Cable and Dr. A. V. 
Hunninen: "Studies in the Life 
History of Siphodera, a Trema- 
tode Parasite of the Toadfish." 

Dr. H. W. Stunkard: "Pathology 
and Immunity to Infection by 
Heterophyd Trematodes." 



FRIDAY, July 18, 8:00 P. M. 
Lecture: Dr. Eric Ponder: "Red 
Cell Structure in the Light of 
Shape Transformation." 



CURRENT APPROACHES TO THE PLANT 
HORMONE PROBLEM 

Dr. George S. Avery. Jr. 
Professor of Botany. 
Connecticut College 
By definition, whether in animals or plants, 
hormones are chemical substances normally pr5^ 
duced in the cells of some part of an organism, 
and transported to other parts 
where in one way or another 
they regulate growth and me- 
tabolism. Of the many fac- 
tors which regnlate growth, 
hormones constitute only one : 
their importance to the devel- 
opment of living organisms 
lies first in the fact that they 
are produced internally (they 
are not ordinarily taken in 
from the environment, for ex- 
ample, as plants take in min- 
erals, etc., from the soil), and 
second, that they exercise their 
regulatory effects when pres- 
ent in minute amounts. 

Plants do not possess secre- 
tory glands, like those of ani- 
mals, but the embryonic re- 

gions such as growing points 

of roots, stems, etc., where 
new protoplasm is constantly being synthesized, 
are the centers of hormone production. 

One of the better understood but not often dis- 



Current Approaches to the Plant Hormone 
Problem, Dr. George S. Avery, Jr 4.5 

The Source of Pancreatic Juice Bicarbonate, 
Dr. Eric G. Ball 45 

The Permeability and the Lipid Content of the 
Erythrocytes in Experimental Anemia, Dr. 
Arthur J. Dziemian 50 



TABLE OF CONTENTS 

Symposia at the University of Chicago 51 

Introducing Dr. Cheng-Kwei Tseng 52 

Items of Interest 53 

Physiology Class Notes 54 

Botany Class Notes 54 

Embryology Class Notes 55 



July 12, 1941 ] 



THE COLLECTING NET 



47 



cussed hormones in higher plants is vitamin Bi. 
thiamin. It is normally produced above ground 
in the green tissues of the plant, and is transport- 
ed to the roots. Root growth, in many species 
at least, cannot go on without it. If roots are 
excised from the parent plant, and grown in cul- 
ture, thiamin has to be supplied in the nutrient 
medium. Thus, in in vitro experiments it is no 
longer a hormone (by definition), but is more 
nearly akin to the growth factors, or accessory 
substances, of microorganisms. I mention this 
only to show that definitions of the past are break- 
ing down as we learn more about the physiolog- 
ically active substances produced by living organ- 
isms. That hormones are "activators" and "cor- 
relating substances" of one sort or another, all 
will agree. Let us leave it at that. 

Before current approaches to the problem are 
mentioned, those of you who are unfamiliar with 
the field will want a little more information, par- 
ticularly about the more commonly discussed 
auxins. Only three auxins have been isolated 
from higher plants : auxins a and b of Kogl, and 
3-indoleacetic acid. A considerable number have 
been synthesized. In spite of the chemical differ- 
ences in the numerous substances known, they all 
bring about the same general non-specific physio- 
logical eff^ects, e.g. they promote growth in length 
of stems, and retard growth in length of roots. 

The history of the discovery of the auxins has 
been treated in detail in "Growth Hormones in 
Plants" by Boysen Jensen et al, and in "Phyto- 
hormones" by Went and Thimann. It is an in- 
teresting record of scientific discovery in which 
the chief object of investigation has been the 
young shoot (coleoptile) of the grass seedling. 
The study of the response of the coleoptile to light 
and gravity has led, over a period of fifty years, 
to the discovery and chemical identification of 
growth hormones in plants. 

Methods of assay. The advance of quantitative 
biology, whatever the special field, usually rests 
on the development of methods for measuring. 
For hormones, chemical methods would be the 
ideal, but thus far no chemical test has been de- 
vised which is sufficiently sensitive to detect any 
of the known plant hormones in the low concen- 
trations in which they naturally occur. Thus, as 
in so many instances in animal physiology, a liv- 
ing test organism is essential. Went gave us the 
Avena (oats) coleoptile test in 1928. It has un- 
dergone a number of modifications since, but ap- 
parently is still the most dependable and most 
quantitative test devised to date. In our labora- 



tory we use the "deseeded" modification suggested 
by Skoog (details can be omitted here). It still 
requires a con.stant temperature constant humid- 
ity darkroom. A test chamber has been developed 
which simplifies greatly the equipment needed for 
making hormone tests, i.e., no humidity control 
is necessary. The "deseeded" Avena test method 
makes possible quantitative estimates of indole- 
acetic acid in concentrations as low as 5 to 10 
micrograms per liter of solution, and with such 
synthetic growth substances as alpha naphtha- 
leneacetic and indolebutyric acids it will give good 
quantitative determinations on concentrations 
down to 25 to 50 micrograms per liter. 

As for the potency of plant hormones : if it 
were physically possible to place a million Avena 
seedlings side by side, with their coleoptiles ac- 
tually touching one another, they would extend 
for a distance of about one mile. And if it were 
possible to cut the tips off all these coleoptiles and 
place tiny agar blocks containing indoleacetic acid 
on one side of coleoptile stump, (as in the Went 
test) one milligram of indoleacetic acid would be 
enough to cause the mile of coleoptiles to grow 
on the side to which the hormone was applied, 
and the final result would be a ten degree curva- 
ture of the coleoptiles (from the vertical posi- 
tion). 

A considerable number of test methods other 
than with coleoptiles have been devised, at least 
one of which will detect much smaller amounts of 
auxin than those just indicated ; the difficulty is 
that it is not very quantitative. Green tissue test 
objects are also in use, but in general they are not 
very sensitive. 

The search for better methods of assay still 
goes on, and in the past year two new ones have 
been developed. Parker-Rhodes proposes a test 
for plant hormones based on osmotic pressure 
changes in root hairs of wheat. This is a wide 
departure from previous methods, practically all 
of which depend upon tissue responses and 
growth curvatures of one sort or another as the 
end result. The other new method is that of Tur- 
fit, who reports, contrary to the earlier studies of 
others, that cell division in yeast is affected by 
both naturally occurring and synthetic auxins ; 
thus he measures increased CO2 output, or after 
a few hours counts cell numbers in yeast cultures 
. . . the increase in cell number being very rough- 
ly proportional to concentration of hormone pres- 
ent in the culture medium. It is too soon to eval- 
uate these methods fairh^ though it is clear that 
they are not free from difficulties. Their adop- 



The Collecting Net was entered as second-class matter .July 11, 1935, at the Post Office at Woods Hole, Mass., 
under the Act of March 3, 1879, and was re-entered on July 23, 1938. It is devoted to the scientific work at 
marine biological laboratories. It is published weekly for ten weeks between July 1 and September 15 from Woods 
Hole, and is printed at The Darwin Press, New Bedford, Mass. Its editorial offices are situated in Woods Hole, 
Mass. Single copies, 30c by mail ; subscription, $2.00. 



48 



THE COLLECTING NET 



[ Vol. XVI, No. 140 



tion by other workers will be the index of their 
success. 

Units. A standard unit of one sort or another 
is as necessary as a good and universally used 
assay method, that is. if workers are to be able 
satisfactorily to compare their results. Some 
progress has been made along this line. Overbeek 
has proposed that hormone content of a tissue be 
expressed in terms of a compound of known 
physiological activity, and has selected indoleace- 
tic acid as that compound. Thus, after assay, one 
may say that the hormone content of a tissue is so 
many gamma (or microgram) equivalents of in- 
doleacetic acid per kilogram of tissue. 

There is one difficulty in the otherwise ideal 
proposal Overbeek has made. He originally sup- 
posed that "gamma equivalents" would be inde- 
pendent of the test method used, but it has since 
been shown that different test methods give dif- 
ferent results with the same hormone extract, i.e., 
two different modifications of Went's method give 
widely different assays when hormone content is 
expressed in terms of gamma equivalents of in- 
doleacetic acid. Thus, for the present at least, 
standardization of units is not possible. 

Getting hormones out of tissue. The ''diffu- 
sion" method was for a long time the only one 
used. This procedure consists of standing a piece 
of plant tissue on a small rectangular plate of agar 
for a standard length of time ; the agar is then 
cut into small blocks, and applied to decapitated 
coleoptiles . . . the Avcna test. This presumably 
gives an index of the hormone concentration in 
the tissue being tested, but does not give any 
quantitative value for the hormone content of tis- 
sue. The necessity for quantitative extraction is 
clear, if we are ultimately to have an understand- 
ing of the role of hormones in growth. 

Here are some of the more recent steps in the 
direction of better extraction methods : Boysen 
Jensen five years ago proposed an ether extrac- 
tion method, whereby the tissue being extracted 
is placed in freshly distilled ether which has been 
freed from peroxides. The tissue is allowed to 
stand overnight in two changes of ether. The 
following day the ether extract of hormone is 
taken down to dryness, and the residue is taken 
up in a small amount of 1.5% agar. Avcna assay 
of the agar-hormone mixture follows. 

It has since been found that if fresh ether is 
added weekly to a sample of tissue from which 
the hormone is 1)eing extracted, that hormone will 
continue to be liberated from the tissue for a per- 
iod of several months, thus suggesting the pres- 
ence of an ether insoluble compound which is 
slowly hydrolyzed into auxin. Thimann and 
Skoog (and others) have struggled with this 
problem of extracting hormone from green tis- 
sues, and have recently reported on a number of 
methods. Some of them gave good yields, but 



with only one tissue were the results to the satis- 
faction of the authors. 

In our laboratory we have sought and found a 
method for total extraction of hormone from non- 
green tissues such as the storage tissues of seeds, 
etc. It give reproducible results and extraction 
takes only a few minutes. It involves heating an 
aqueous suspension of the tissue at 100 to 120 
degrees C. for fifteen minutes, at a pH of 9 to 10. 
After heating, the suspension is centrifuged and 
the pH of the clear extract is adjusted to approx- 
imately 6 ; agar blocks are prepared from this 
aqueous extract of the hormone, and assayed by 
the Avcna method. Corn endosperm extracted 
by this method gives yields as high as the equi- 
\'alent of 150 milligrams of indoleacetic acid per 
kilogram of tissue (content varies according to 
variety). This is a higher yield than has yet been 
reported, and is due to the conversion of a "pre- 
cursor" compound into auxin. The auxin pres- 
ent in such great quantities in maize endosperm 
has been shown to be indoleacetic acid, and the 
chemical identity of the precursor is just about to 
be established. 

These discoveries of the past few months mark 
an important advance in the extraction and iden- 
tification of naturally occurring physiologically 
active substances. Less recent, but equally im- 
])ortant, are the isolation and identification of 
wound and leaf growth hormones at the Califor- 
nia Institute of Technology. Auxins a and b are 
now an old story and ma\' turn out to be less im- 
portant than originally thought. The chief point 
is that the plant hormone family is increasing in 
number, and in addition to those growth promot- 
ing hormones already mentioned, it seems likely 
that one or more substances specifically concerned 
with differentiation may soon be isolated and 
identified, e.g., flower and sex differenting sub- 
stances. 

Hormones and normal growth. Methods of 
extraction and assay would be of little value if 
they could not be applied to problems of growth 
and differentiation. What evidence is there for 
auxins being agents which exercise developmental 
control in organisms? Here, in brief, are the re- 
sults of a few studies : It has been shown in cer- 
tain leaves that the regions of greatest growth in- 
tensity are also the regions of highest auxin con- 
centration : that plant shape and rate of develop- 
ment of plant organs may be correlated with 
auxin concentration ; that the "cambial stimulus" 
in trees (initiation of lateral growth in the spring) 
is related to auxin concentration ; that dwarfing 
in maize is related to destruction of auxin ; that 
"lazy" maize, which as its name implies falls over 
and grows on the ground, gets its lazy habit at 
least in part from mal-distribution and mal-trans- 
port of auxin in its tissues. Other examples might 
be cited, but this is a quiet sector on the hormone 



July 12, 1941 ] 



THE COLLECTING NET 



49 



front just now. Better extraction methods must 
be developed before too much faith is placed in 
studies on growth in relation to hormone content 
of tissue. 

Syntlietic plant growth substances. For all 
practical purposes, we might as well call these 
substances "hormones" also. Most of them were 
first reported by Zimmerman and Wilcoxon in 
1935. A few of them (indoleacetic acid is now 
in the "naturally occurring" group for higher 
plants), in the approximate order of their poten- 
cy, are alpha naphthaleneacetic acid, indolebutyric 
acid, indolepropionic acid, phenylacetic acid, etc. 
A newcomer, beta naphthoxyacetic acid, is bid- 
ding for an important place ; in addition to 
bringing about responses in the usual curvature 
and other tests, it will also induce form changes 
if sprayed on plants in high concentrations (or if 
the plants are watered with it). Except for the 
preliminary work of Pearse, this is the first com- 
pound of a hormone nature with which it has 
been possible to demonstrate morphogenetic ef- 
fects in a living intact plant ; needless to say, this 
marks an important advance. 

Horticultural applications. The use of syn- 
thetic hormones in the rooting of cuttings of high- 
er plants is probably the best known of all the 
new practical applications, and "rooting com- 
pounds" are on sale in almost all seed stores. 
When sprayed on, or otherwise applied to flowers, 
under the proper circumstances, seedless fruits 
are developed . . . and such seedless tomatoes, I 
am told, are in commercial production. Egg- 
plant, squash, peppers, cherries and even orna- 
mental holly berries are now on the potential 
"seedless" list, but worse, watermelon tradition 
is going to be changed . . . for it is possible to 
have seedless watermelons too. 

Perhaps the most important horticultural use 
thus far is spraying these substances on apples 
etc. to prevent pre-harvest drop of fruit. The 
manner in which hormones prevent abscission is 
not yet understood, but that they are abscission- 
controllers, there is no doubt. 

Hormones in relation to abnormal growth. 
Not new, but of increasing significance, is the 
work of Kraus ct al at the University of Chicago. 
He and his coworkers have applied high concen- 
trations of synthetic hormones to bean, and other 
species, and studied the tissue changes which 
followed. Ultimately the overgrowths produced 
develop into good-sized galls. They are not "can- 
cers", but are perhaps as cancer-like as anything 
which plants can produce. Chemical analysis of 
the treated tissues indicate that the applied hor- 
mones are responsible for mobilizing nitrogen, 
carbohydrates, etc., at the place of application. 
This is very suggestive. Is it possible that animal 
cancers secrete mobilizing substances, i.e., that 
once started they continuously secrete such sub- 



stances, thus perpetuate themselves? 

Treatment of sunflower and rose stems with 
certain of the animal carcinogens has failed thus 
far to bring about the response just noted for 
plant hormones, but the problem needs re-ex- 
amination with younger test material which might 
have greater capacity to respond. Of course it 
may be that the water insolubility of these com- 
pounds makes it impossible for plant cells to take 
them in in quantities sufficient to bring about a 
response. 

The spontaneous tumors occurring in the hy- 
brid of Nicotiana glauca X ^. langsdorjii de- 
serve attention in passing. 

The role of hormones in the production of 
plant tumors is as yet but little understood. It is 
important for a general understanding of growth 
that we go further into the problem. Alert stu- 
dents will find a fertile field. 

Possible roles of hormones in plant grozvth. 
Mention of mobilization efifects attributable to 
hormones has just been made, but it seems un- 
likely that the primary eft'ect of hormones would 
be on mobilization. Critical studies of hormone 
influence on metabolic rates must be made, and 
extended studies on hormone-enzyme relation- 
ships should prove very fruitful ; on this latter 
point there are some interesting suggestions al- 
ready. It has been known for some time that 
auxin is present in higher concentrations in the 
tip than elsewhere in the Avena coleoptile, and 
recent studies of peptidase distribution in the co- 
leoptile show that peptidase activity is also higher 
at the tip. Is auxin acting as an enzyme activa- 
tor, as has been suggested? Further promising 
work along this line has been done by Commoner 
and Thimann, who give limited evidence of auxin 
activation of certain events in the respiratory 
chain. 

The fact that different auxins can bring about 
the same general physiological responses in plants 
(whatever specificity there is, residing in the 
species) is analagous to the coenzyme-enzyme re- 
lationship, where a single substance is known to 
act as a coenzyme for a number of enzymes, and 
the specificity of the combination resides in the 
enzyme. 

Although not often discussed along with the 
auxins, thiamin is the one plant hormone of 
which at least one role is understood. It has 
been shown in in vitro experiments to act as a 
cocarboxylase. There is nothing in the chemical 
structure of the various auxins to indicate whe- 
ther they, like thiamin, may be acting as coen- 
zymes, but the preliminarj' evidence favors the 
view that they act as enzyme activators, or com- 
ponents of enzyme systems. 

The really important problems remain to be 
solved. We know little about the production of 
such physiologically active substances by organ- 



50 



THE COLLECTING NET 



[ Vol. XVI, No. 140 



isms, or how to extract and identify them. And 
we need much more information on the destruc- 
tion of such substances in hving tissues, more on 
inhibiting- suljstances of one sort or another, and 
most of all, we need to know what role they play 
in growth and development. 

Growth factors for microorganisms. Now just 
a little material to correlate the foregoing with the 
studies being made on the growth factors or ac- 
cessory substances for microorganisms. 

Certain microorganisms do not possess the 
ability to synthesize optimal amounts of these 
growth factors, and it is by supplying such defi- 
cient substances through the culture medium that 
we have learned about the necessity for them. 
Some of the substances in question are known to 
act as coenzymes or parts of coenzyme systems 
(thiamin, nicotinic acid, etc.), and it appears as 
if their roles in the growth of microorganisms 
were analagous to those of auxins and other hor- 
mones in higher plants. 

From the microorganism work have come some 
interesting results, from the viewpoint of helping 
us to understand symbiosis and parasitism. Kogl 
and Fries, for example, grew two totally unre- 
lated fungi in the same culture. One produced 
biotin, needed by the other for satisfactory 
growth while the other produced thiamin, needed 
by the first for good growth ; the result was sym- 
biosis in vitro. 

A suggestion for the future. How may such 
information spur new research approaches on the 
relation of physiologically active substances to 
growth and development ? Lichens may well be 



the plants we ought to study in this connection. 
They are compound organisms which exhibit 
symbiosis. One of the component organisms, the 
alga, synthesizes food which it delivers to the 
fungus component, while the latter supplies min- 
erals and water to the alga. This relationship has 
been explained in the past as a perfectly straight- 
forward nutrient partnership. The situation now 
seems less simple : it is undoubtedly more than a 
nutritive bond which holds an alga and fungus 
together, and produces an organism with new 
characteristics, an organism which is more than 
the sum of its parts. So stable is this relation- 
ship that the resulting plant bodies require gen- 
eric and specific names. Nowhere else in the 
plant kingdom, so far as I know, do two com- 
pletely different organisms live in such intimate 
relationship and produce specific new physical 
forms. There are no genes for lichens, and nutri- 
tion offers only limited possibilities of explaining 
the mode of their existence ... it contributes 
nothing to an explanation of the new physical 
form assumed. Thus it appears that each com- 
ponent is secreting one or more substances of a 
"growth factor" nature which are influential in 
determining the form of the dual organism. If a 
lichen were a single organism we would say that 
its form is dependent upon its genetic constitu- 
tion, its genes. There is a good likelihood, it 
seems to me, that growth factor studies on the 
lowly lichen may well provide a means of getting 
at the problem of the chemistry of the gene. 

(This article is based upon a lecture delivered at 
the Marine Biological Laboratory on July 3.) 



THE PERMEABILITY AND THE LIPID CONTENT OF THE ERYTHROCYTES IN 

EXPERIMENTAL ANEMIA 

Dr. Arthur J. Dziemian 
Department of Physiology, School of Medicine, University of Pennsylvania 



Bodan.sky. (/. Biol. Chcm., 63:239,1935) re- 
ported, a number of years ago, that administration 
of phenylhydrazine and allied compounds to ani- 
mals brought about changes in the lipid content 
of the red blood cells. This paper describes ex- 
periments on the efifect of phenylhydrazine on the 
permeability of the erythrocyte and its lipid con- 
tent. 

Albino rabbits were used in all these experi- 
ments and phenylhydrazine hydrochloride solution 
was injected subcutaneously. In five animals 
changes in the red cell counts, hematocrits, reticu- 
locyte percentages, erythrocyte diameters, and per- 
meability of the red cells to glycerol, diethylene 
glycol, ammonium propionate and ammonium sa- 
licylate were followed during the onset of and re- 
covery from phenylhydrazine anemia after a single 
dose of the drug. 



The number of red cells decreased rapidly, so 
that about the sixth day after treatment, the count 
was only about one quarter of the original value. 
After the sixth day the number of erythrocytes 
slowly returned toward normal. Hematocrit 
changes followed the variation in red cell count 
rather closely, with differences due to changes in 
the size of the erythrocytes. 

About the second day after treatment, the retic- 
ulocyte percentage increased rapidly, new cells 
coming in to replace the original phenylhydrazine 
poisoned red cells. The percentage rose to be- 
tween 30 and 40% of the total number of red cells 
on the seventh day, and then dropped back to the 
normal value of below 1% by the 16th day after 
administration of phenylhydrazine. 

The mean diameter of the erythrocytes de- 
creased for the first few days of the experiment, 



July 12, 1941 ] 



THE COLLECTING NET 



51 



but as the new reticulocytes entered the circula- 
tion, the average size of the cells was considerably 
increased above normal. The reticulocytes are of 
large size, some being eleven micra in diameter, 
as compared with a normal average of 6.8 micra. 
.^fter the 9th da}' the new cells began to assume 
normal size slowly. Comparisons of frecjuenc}' 
distribution curves of the diameters of the cells 
on various days after treatment showed that by 
the 8th day practically all the original shrunken 
cells were gone, indicating an almost complete 
new population of erythrocytes. 

Permeability of the red cells was measured by 
rates of hemolysis of the erythrocyte in buffered 
solutions (pH 7.4) of the penetrating substances 
at 20 degrees C. The time to 50% hemolysis was 
taken as a measure of permeability. 

Phenylhydrazine HCl in vitro has no effect 
upon the permeability of the erythroc3'te, nor does 
it have any hemolytic effect, even after the cells 
are in contact with it for several days at 38 de- 
grees C. The theory of its action in vivo is that 
the hemoglobin of the cell is changed in part to 
free hemin and reduced globin, which catalyzes 
the change of the rest of the hemoglobin to 
methemoglobin. Erythrocytes containing methe- 
moglobin are supposed to be more readily attacked 
by the reticulo-endothelial system. 

The rate of penetration of glycerol into the 
erythrocyte of the treated rabbits increased great- 
ly, being fastest about the 8th day after injection 
of phenylhydrazine. It then became slower, tak- 
ing about 80 days to reach the original value. In 
some cases the erythrocytes of the treated animals 
were from 20 to 29 times as permeable as the 
erythrocytes of the same rabbits before adminis- 
tration of the drug. Diethylene glycol penetration 
followed much the same course as did glycerol. 

For the first couple of days after treatment, the 
erythrocytes showed an increase in permeability 
to ammonium propionate and salicylate, which 
paralleled the decrease in cell size. But as new 
cells came into the blood, the time to 50% hemo- 



lysis increased above the original value, and re- 
mained above normal for several weeks. Whereas 
the young cells of the rabbit were more permeal:)le 
to glycerol and diethylene glycol than were the 
normal erythrocytes, they were less permeable to 
ammonium salts of organic acids, the so-called 
lipid-soluble substances. 

The erythrocytes of eight normal albino rabbits 
were analyzed for total lipid, cholesterol, phos- 
pholipid, and neutral fats by gasometric methods. 
The permeability of these cells were also ascer- 
tained. The animals were then injected with 
]3henylhydrazine hydrochloride, and the determin- 
ations were repeated. Blood was taken at various 
times after treatment, in some cases when the 
blood contained a high percentage of reticulocytes 
and in others after the rate of inflow of immature 
cells had decreased. In all cases the amount of 
total lipid per erythrocyte increased, but so did 
the area of the erythrocyte. When the total lipid, 
cholesterol, phospholipid and neutral fat contents 
were calculated per unit area of cell surface, there 
was no regularity, the distribution seemed random 
and not outside normal variation. 

The results on permeability showed that the re- 
sistance of the treated red cell decreased in every 
case studied to glycerol and diethylene glycol. 
The figures for 50% hemolysis of the erythrocytes 
of the treated animals in the ammonium salt solu- 
tions indicated that they were taken at different 
times during the onset of and recovery from the 
anemia. In some the penetration was faster, the 
blood having been taken shortly after treatment, 
and in others, slower, having been drawn about 
ten to twelve days after injection. 

No correlation seems possible with these re- 
sults. They indicate that it is not the amount of 
lipid in the red cell surface that controls the per- 
meability of the erythrocyte to the penetrating 
substances studied. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on July 
8.) 



SYMPOSIA AT THE UNIVERSITY OF CHICAGO 



In connection with the fiftieth anniversary cele- 
bration of the University of Chicago at the end 
of September, and a special meeting of the Amer- 
ican Association for the Advancement of Science 
convening there, symposia will be held in the va- 
rious fields of human learning. 

The symposia scheduled in the field of biology 
are : 

"Growth and Differentiation in Plants" — Chair- 
man, Ezra J. Kraus. Speakers: Charles E. Allen, 
University of Wisconsin; Edmund W. Sinnott, Yale 
University; John W. Mitchell, U. S. Department of 
Agriculture; John M. Beal, University of Chicago. 

"Levels of Integration in Biological and Social 
Systems" — Chairman, William H. Taliaferro. Speak- 



ers: Libbie H. Hyman, American Museum of Na- 
tural History; James W. Buchanan, Northwestern 
University; Herbert S. Jennings, University of Cali- 
fornia at Los Angeles; and Ralph W. Gerard, Wil- 
liam Burrows, Thomas Park, and Warder C. Allee, 
University of Chicago. Special lecture: Donald D. 
Van Slyke, Rockefeller Institute for Medical Re- 
search. 

"Visual Mechanisms" — Speakers: Selig Hecht, 
Columbia University; Ernst Gellhorn, University of 
Illinois; Samuel H. Bartley, Washington Univer- 
sity; Karl S. Lashley, Harvard University; and 
Arlington C. Krause, Heinrich Kluver, Theodore J. 
Case, and Stephen Polyak, University of Chicago. 

"Levels of Integration in Biological and Social 
Systems" — Chairman, Robert Redfield. Speakers: 
(Continued on page 52) 



52 



THE COLLECTING NET 



[ Vol. XVL No. 140 



The Collecting Net 

A weekly publication devoted to the scientific work 
at marine biological laboratories. 

Edited by Ware Cattell with the assistance of 
Boris I. Gorokhoff and Judy Woodring. 

Entered as second-class matter, July 11, 1935, at 
the U. S. Post Office at Woods Hole, Massachusetts, 
under the Act of March 3, 1879, and re-entered, 
July 23, 1938. 

3(^tl•o^urinl^ 

Dr. Cheng-Kwei Tseng, Assistant Professor of 
Botany and Acting Curator of Herbarium. Ling- 
iian University, China ; LTniversity Fellow in Bot- 
any, LTniversity of Michigan. 

A native of Amoy, China, Dr. Tseng has spent 
most of the last ten years collecting marine algae 
along the entire coast of China. He has worked 
on all phases of Chinese algology. including tax- 
onomy, morphology, ecology and economic uses, 
and has published a number of papers in Oriental 
botanical and scientific journals on these subjects. 

Upon completing his post graduate studies at 
Lingnan University in 1934, he was appointed 
lecturer in botany and curator of the herbarium 
at the University of Amoy. A year later he was 
made assistant professor of botany and curator 
of the herbarium at the National University of 
Shantung. Tsingtao, positions which he held un- 
til 1938, when he was appointed to Lingnan Uni- 
versity. During ]iart of this time he was collab- 
orate algologist at the Marine Biological Station 
in Amoy. In 1937 he was connected with the 
newly established Marine Biological Station at 
Tsingtao, but the station was destroyed by the 
Japanese soon afterwards. 

Last September he arrived in the LTnited States 
to work at the University of Michigan under Dr. 
\Xm. Randolph Taylor. He is particularly inter- 
ested in correlating species of Chinese algae with 
those of the United States coasts. He considers 
it quite likely that a number of Oriental species 
of algae, which have been given separate specific 
names, are actually identical with American 
species, and, conversely, that species hitherto con- 
sidered to be the same may actually be different. 
The only way to resolve these questions is to 
study specimens from both the Pacific and the 
Atlantic regions, preferably in the living condi- 
tion. He has studied Chinese forms extensively, 
and is now working on certain American species 
for the purpose of comparison. In connection 
with this work, he brought part of his algae col- 
lection, amounting to about 3,000 specimens, with 
him to the United States. 

He has also been working on monographs of 



certain Chinese algae, and has already published 
since his arrival in this country treatises on Lia- 
gora, WrangcUa, Griffitlisia. Codiiim and Cliac- 
tangiaceae. 

At Woods Hole this summer he is also assist- 
ing in the Botany course, together with Mr. W. 
J. Gilbert, taking the place of Dr. Rufus H. 
Thompson. Dr. Tseng will spend the academic 
year 1941-42 at the University of Michigan and 
will then return to China. 

ADDITIONAL INVESTIGATORS 

Aquila, (Sister) M. grad. biol. Villanova. Rock 3. 
Bodian, D. asst. prof. anat. Western Reserve Med. 

lib. 
Brown, D. E. S. prof. phys. New York. Br 310. 
Calkins, G. N. prof, proto. Columbia. Br 331. 
Chambers, E. New York Med. Br 343. 
Chambers, R. prof. biol. New York. Br 328. 
Duncan, G. W. fel. surg. Hopkins. Br 328. 
Genevieve, (Sister) Mary grad. biol. Villanova. Rock 

3. 
Gilbert, P. W. instr. zool. Cornell. OM. 
Lancefield, D. E. assoc. prof. biol. Queens (New 

York). Br 126. 
Pick, J. instr. anat. New York Med. Br 343. 
Rahn, H. instr. zool. Wyoming, lib. 
Stebbins, R. B. grad. asst. biol. New York. Br 328. 
Stiegelman, S. asst. zool. Columbia. Br 313. 

SYMPOSIA AT THE UNIVERSITY OF CHICAGO 

(Continued from page 51) 

Clarence R. Carpenter, Pennsylvania State College 
and the School of Tropical Medicine (Puerto Rico); 
Alfred L. Kroeber, University of California; and Al- 
fred E. Emerson and Robert E. Park, University of 
Chicago. 

"Sex Hormones" — Chairman, Frank R. Lillie. 
Speakers: Edward A. Doisy, St. Louis University; 
John S. L. Browne, McGill University; and Carl R. 
Moore, Allan T. Kenyon, and Fred C. Koch, Univer- 
sity of Chicago. 

"Immunological Mechanisms" — Chairman, George 
F. Dick. Speakers: Linus Pauling, California In- 
stitute of Technology; Thomas M. Rivers, Hospital 
of the Rockefeller Institute; and William Bloom, 
Paul R. Cannon, and William H. Taliaferro, Univer- 
sity of Chicago. Special lectures: Charles H. Best, 
University of Toronto; and Ernst W. Goodpasture, 
Vanderbilt University. 



CURRENTS IN THE HOLE 

At the following hours (Daylight Saving 
Time) the current in the Hole turns to run 
from Buzzards Bay to Vineyard Sound: 
Date A. M. P. M. 

July 12 7:00 7:21 

lul'v 13 7:47 8:10 

July 14 8:35 9:01 

Tulv 15 9:23 9:55 

Tulv 16 10:15 10:50 

Tulv 17 11:07 11:45 

July 18 11:58 

In each case the current changes approxi- 
mately six hours later and runs from the 
Sound to the Bay. 



July 12, 1941 ] 



THE COLLECTING NET 



53 



ITEMS OF INTEREST 



Dr. and Mrs. Charles Packard will be at 
home to membei-s of the Laboratory on Sunday, 
July 13, and on the two following Sundays, from 
four-thirty to six o'clock. 

Dr. Robert George Ballentine was granted 
a National Research Council Fellowship for the 
3^ear 1941-42 to work on the chemical organiza- 
tion of the cell surface at the Rockefeller Institute 
for Medical Research in New York. He is as- 
sisting in the physiology course at the Marine 
Biological Laboratory. 

Dr. Margery Milne has been appointed as- 
sistant professor of biology in the Richmond Di- 
vision of the College of William and Mary. 

Dr. Dorothy M. Wrinch, the English mathe- 
matician and theoretical biochemist, has been en- 
gaged jointly by Smith, Mt. Holyoke, and Am- 
herst Colleges to conduct at each institution next 
year a series of seminars open to the members of 
the three faculties and their advanced students. 
W^orking under a grant from the Rockefeller 
Foundation, she is at present a member of the 
faculty of physical science at Oxford and a lec- 
turer in chemistry at the Johns Hopkins Univer- 
sity. 

Dr. Eugene _F. DuBois has been appointed 
professor of physiology at Cornell University 
Medical College, and head of the department of 
])hysiology and biophysics, not biochemistry and 
physiology as previously reported. 

Dr. John H. Northrop, of the Rockefeller In- 
stitute of Medical Research, received the honorary 
degree of Doctor of Science at Rutgers Univer- 
sity on June 8. 

Next week the physiology class will have the 
following guest speakers : Dr. M. E. Krahl will 
speak on Thursday, and Dr. O. C. Glaser will talk 
on "General Physiological Problems of Growth" 
next Saturday. Guest lecturers who have spoken 
previously are Dr. L. V. Heilbrunn, whose topic 
was "Protoplasmic Viscosity" and Dr. A. C. Red- 
field who lectured on "Respiratory Proteins." Dr. 
E. G. Conklin lectured before the embryology 
class today on the mapping of eggs. 

Dr. Alma G. Stokey, of Mount Holyoke Col- 
lege, will give a paper at the botany seminar on 
Thursday, July 18. She will talk on "The Game- 
tophytes of the Filmy Ferns." Last week Dr. F. 
B. Smith of the University of Florida spoke on 
"Types and Distribution of Microorganisms in 
Some Florida Soils." 



Dr. John S. Rankin, instructor in the inver- 
tebrate zoology course at the Marine Biological 
Lal)oratory, is being married to Miss Julia Pen- 
field Smith, (Invertebrate, 1940), today in Rock- 
ford, Illinois. 

Miss Muriel Voter, an instructor at Whea- 
ton College, married Carroll Williams, next year 
a Harvard Fellow, on June 26th in Middlebury. 
Vermont. Both the bride and groom took the 
class in Invertebrate Zoology at the Laboratory 
in 1939. 

Mr. David J. Bradley, son of Dr. and Mrs. 
Harold C. Bradley, was married on April 26 to 
Miss Elizabeth B. McLane in Manchester, New 
Hampshire. The couple will make their home in 
^Madison, Wisconsin. 

Dr. Roberts Rich and his family will be in 
New York this summer. Dr. Rugh is teaching a 
course in embryology in the summer school of the 
Washington Square College of New York Uni- 
versity. 

Mr. W. R. Dillon, chief of the division of 
administration of the Fish and Wildlife Service, 
is visiting the station with his wife and son for 
the month of July. They are staying at the Fish- 
eries residence. 

Firms which held exhibits at the Marine Bio- 
logical Laboratory during the past week included : 
The Macmillan Company (Mr. Harvey C. McCa- 
leb) ; The Spencer Lens Company (Messrs. 
Charles Riley and Frank Munoz) ; and Ward's 
Natural Science Establishment. 

Choral Club rehearsals are continuing on Tues- 
day and Thursday evenings. An appeal has been 
issued for more basses and tenors. On Thursday 
night these parts were filled by a group of 
soldiers from the 181st Regiment, who, on the 
spur of the moment, joined in the singing. 

The program of the phonograph record concert 
next Monday at the M.B.L. Club will be as fol- 
lows : Handel. "Concerto in D for orchestra, with 
organ" ; K. P. E. Bach. "Concerto in D for or- 
chestra" ; J. S. Bach, "Brandenburgh Concerto 
No. 6"; intermission; Gluck, (opera) : "Orpheus 
and Eurydice." 

The M.B.L. Tennis Club has appointed Dr. E. 
R. Jones as treasurer until this summer's elec- 
tions, to replace Dr. T. K. Ruebush, who was 
called into service in the U. S. Navy. Dr. D. E. 
Lancefield, club president, announces that the 
beach courts are now available for playing. 



54 



THE COLLECTING NET 



[ Vol. XVL No. 140 



PHYSIOLOGY 

With our last week of lectures and supervised 
lab work just about over, we are all busily en- 
gaged in the scheming of some brilliant and rare 
bit of research with which to occupy ourselves 
next week. The problem of finding a problem is 
difificult enough in itself, but already some of us 
have reached a decision. Fred Coe. being a great 
peanut enthusiast, is considering the problem of 
peanut Ijutter metabolism in Crustacea. Bernie 
Shepartz, who suggested this to Fred, will assist 
in the experiments by first determining whether 
the Crustacea like peanut butter or not. Frank 
Hartman, emerging from the depths of the cellar 
where he has been micro-manipulating all week, 
thinks he will study tissue regeneration, his ap- 
paratus for which will consist of one good bed, 
plenty of time, and an alarm clock set for supper. 
Ed Burns has become quite interested in the 
work with dogfish ghosts and so he plans to 
study the art of ghost-ljreaking with the aid of the 
Topper series as reference. 

Just a word about our favorite delicacy — micro- 
manipulation. At the beginning of the week we 
handled the needles and pipettes as if we had re- 
ceived our earl\' training stacking cord wood. By 
the end of the week at least a few members 
emerged from beneath a stack of broken needles 
with a triumphant air. proudly holding samples of 
their work. We enjoyed the work very much 
even if we did have to get down to such fine 
points. 

On Tuesday we changed sections and each one 
got involved in something new. The second 
group always gets a break because the apparatus 
is usually still set up and at least one big head- 
ache is thereljy eliminated. There also seems to 
be an advantage because advice is available from 



CLASS NOTES 

those who have gone before and know what not 
to do. 'i'husly have the new micro-manipulators 
learned that dirty needles and clogged pipettes 
cannot he cleaned with a handkerchief, while the 
new Van Slykers are still wondering about the 
Ijest method for gathering up mercury from the 
floor. The new cytochemists received a list of 
eighteen "don'ts" made out by their passing fel- 
lows and Dr. Ballentine. It seems both the stu- 
dent and Dr. Ballentine had a little trouble, so the 
first "don't" reads : — "Don't break glassware, 
especially burettes (instructor please copy)." 

Tuesday turned out to be a lovely (?) day for 
a picnic, so we found ourselves, as usual, listen- 
ing to a lecture at 9 A. M. Dr. Fisher, while 
gazing out the window, was heard to say, "I'd 
even rather work than go on a picnic today." So 
the lobsters were placed in a tank where they 
fought all day, the rest of the food was hidden, 
and five minutes were appropriated to silent 
prayer for sunshine on \\^ednesdav. 

Dr. Kempton delivered the lecture, his subject 
being observations made from direct investiga- 
tions of glomerular and tubular renal functions. 
On Thursday he spoke on "The Concept of Renal 
Clearance." and on Friday, "The Problem of 
Tubular Secretion in the Kidney." Dr. A. C. 
Redfield was our guest lecturer on Saturday, his 
topic being "Respiratory Proteins." 

Many mistakes which we might have made this 
week were avoided through the kind ofifices of an 
c.r-officio physiolog}' instructor who managed to 
find time, despite the exigencies of his embryol- 
ogy course, to instruct us in all phases of physiol- 
ogv. \ye thank him. — /. E. H. 



BOTANY CLASS NOTES 



Last Sunday, Dr. Runk spread a huge quantity 
of delectable foods before a group of the younger 
botanists on their visit to his home in Nantucket. 
That trip was the highlight of the week — probably 
of the course, too — and will doubtless be remem- 
bered by everyone for a long while to come. Being 
invited to spend the entire day on the island, the 
class left on the early boat, according to plans 
made by the ever-efificient Nancy Bull. 

Upon docking, their first experience was an in- 
side view of Maria Mitchell's house, where the 
telescope \\ith which she discovered her comet is 
still in its old place. A pleasant surprise was the 
display of fresh Nantucket flowers on exhibit 
there. After this, the group left in cars for Dr. 
Runk's house, three brave souls — Connie Stanton. 
Jean Enzenbacher and Bob Thorne — taking a 
wild ride via the dunes and moors, where the 
driver tried to prove that travel is much more ex- 
citing when one doesn't bother with roads. 

Swimming, talking, and then the boatride home 



with that gorgeous sunset in the West was truly 
the end of a perfect day for the tired algologists 
who finally reached Woods Hole in the evening. 

A notable casualty next day was Connie Stan- 
ton's jaiix-pas in spilling a new bottle of Scrip ink 
over the desk, herself, and the general upper end 
of the Botany lab. while Dr. Taylor, unperturbed 
at the mishap, calmly continued his lecture. 'Tis 
said the lady demonstrated admirable nonchalance 
by placidly powdering her nose, but that's unadul- 
terated gossip. 

Not being lazy, as are certain unmentionable 
classes at the M.B.L.. the botanists spent the 
whole damp Fourth in getting acquainted with 
marine algae at Nonamesset Island and Spindle 
Rock. In case any morose individual needs cheer- 
ing, the guaranteed remedy is a position on the 
docks somewhere watching the chain of six skiffs 
slowlv towed through the Gutter amid cheers and 
catcalls from the Nantucket. Unanimously pro- 
claimed the best trip yet, this excursion listed only 



July 12, 1941 ] 



THE COLLECTING NET 



55 



two interesting accidents — Babs Bayard's fatal 
slip at the end after so carefully keeping semi-dry, 
and Sam Salvin's graceful misstep on Mytilus, 
which lost him his whole bucket of algae, only 
part of which he retrieved. Mild excitement was 
caused when Jackie Waldron and Connie Stanton, 
rowing one skiff, tried to shanghai "C.-K." Tseng 
and Bob Williams by heading for New Bedford 
instead of Eel Pond. They soon decided, though 
that the project wasn't worth the effort, and then 
headed for home like good children. 

Last Wednesday, the class took another trip on 
the Nereis, that time to Nashawena. Pasque and 

EMBRYOLOGY 

Academic Life: 

The past week has been spent on Echinoderms 
with Dr. Schotte, who dwelt at first on the illu- 
sive orange ring of the eggs of Paracentrottis. The 
structure of Arbacia eggs was explained, and their 
polarity. It was shown by various experiments 
the lack of relationship between cleavage patterns 
and the final development of the egg. In the 
middle of the week we had a lecture' by Dr. 
Chambers, renowned for his work in microdissec- 
tion. The removal of nuclear material from eggs 
is as easy for him as an appendectomy is for a 
surgeon. This particular lecture was on activa- 
tion of eggs and the surprisingly small part the 
sperm plays in the whole process of fertilization. 

We went on with Dr. Schotte on artificial par- 
thenogenesis. The class as a whole obtained quite 
good results, showing that the female can carry 
on. For further reference, Ray has suggested 
that we read "This New Magnificent World," by 
Huxley, in which the future of the Test Tube 
Embryos can be utilized as our first line of de- 
fence. 

Dr. Schotte inspired us with his talks on the 
regeneration of amphibian limbs, which has been 
his special field during recent years. He is plan- 
ning to test his theories on mammals as he feels 
that some aspects of his field can be applicable 
to higher vertebrates. As an example, the trans- 
plantation of periostial tissue will regenerate bone. 

Social Life : 

As mentioned in our last issue, our soft-ball 
prowess is improving, as was further shown when 
the crew defeated us by only two runs, quite a 
contrast to our first game with them. If a week's 
active training did this, just think what another 
week will do. Time will tell the tale. (Editorially 

THE SOURCE OF PANCREATIC JUICE BICARBONATE 

(Continued from page 45) 
when the gland works hardest it produces the lyzes the reaction CO^ + HoO 
most COo and this is reflected in a higher bicar- 
bonate content of the juice. Second, because the 
pancreas is one of the few tissues that contain the 
enzyme carbonic anhydrase. This enzyme cata- 



Naushon Islands. Pasque, it seems, proved the 
most exciting for Bill Gilbert's group, who were 
broad-minded enough in their search for algae to 
sto]i to appreciate a pair of deer thnt approached 
them fairly closely before gracefully bounding off 
again. Some of the city critters h.id never seen 
wild deer — imagine ! 

Finally and foremost, the week's activities were 
climaxed last Monday evening by some more of 
Dr. Taylor's color film — gorgeous enough to make 
any true botanist realize he'd not have to be an 
esthete, even, to appreciate his algal materials. 

— /. W. 

CLASS NOTES 

speaking we hope it will be so— we disclaim re- 
sponsibility.) 

A peculiar, but very familiar odor greeted us 
one morning as we came from breakfast. Appar- 
ently there had been a skirmi'-h in back of Physi- 
ology. The unmistakable results were obvious : 
They suffered ! ! 

The Glorious Fourth was ushered in on the 
third and finished up on the fifth, at the picnic. 
The Embryology Lab was blitzkrieged from the 
roof of the Brick Building by some belligerent 
soul who objected to the industrious way we were 
spending our National Holiday (we had a radio 
— even an inspiration for dancing — remember, 
Trink and George). 

The climax of our social life reached a peak 
Saturday with the Embryology PICNIC at Tar- 
paulin Cove ! We took 'Winnie' and Sagitta, or 
rather they took us the long way 'round amid 
songs and merriment. The duel of boats between 
Stirling and Fred (being towed) was a jumping 
off point of hilarity for the rest of the day. Clams, 
lobsters and Dr. Goodrich's scientific carving of 
watermelon formed the nucleus of our excellent 
and filling meal. Carrying out the theory that 
fingers were made before forks simplified the 
mechanics of eating. Trink and Tom chose sides 
for soft ball in which the sea played an important 
part as left field. Much fun was had by all. A 
sleepy, sunburned crowd finally piled into the 
boats. The Pirates of the Sagitta stormed 'Win- 
nie's' stronghold, carrying off what food remained. 
Trink was the star performer — with two cans of 
beer in one hand and four sandwiches in the 
other. At that he almost missed the boat. Our 
hearts and hands to you — Neil — for a grand pic- 
nic — efficiently organized. — P. and K. 



H0CO3. Now 
if the metabolic CO2 of the gland is to be con- 
verted to bicarbonate for the juice, the first step 
in the process would be its hydration according 
to the above equation. Thus the occurrence of 



56 



THE COLLECTING NET 



[ Vol. XVL No. 140 



carbonic anhydrase in the pancrea.s may be looked 
upon as an indication that this enzyme is needed 
there in order to speed up the hydration of meta- 
bolic COi; for the production of bicarbonate for 
the juice. If carbonic anhydrase plays such a 
role in the pancreas then any changes in its activ- 
ity should be reflected in the composition or the 
rate of flow of the pancreatic juice. Mann and 
Keilin have recently shown that the drug sulfani- 
lamide is a specific and potent inhibitor of car- 
bonic anhydrase activity. We therefore under- 
took a study of the effect of sulfanilamide injec- 
tions on the composition of the pancreatic juice 
of dogs. Somewhat to our surprise no effect on 
the rate of flow or the bicarbonate content of the 
juice was oliserved even when the concentration 
of sulfanilamide in the juice itself reached a value 
of 64 mg%. This concentration is 200 times that 
needed to completely inhibit the enzyme in vitro. 
These results therefore indicated that carbonic an- 
hydrase had nothing to do with production of pan- 
creatic juice and suggested that the metabolic 
CO-2 of the gland was not an important source of 
the juice bicarbonate. 

If this was the case then the bicarbonate of the 
juice must be derived mainly from the blood 
stream. In order to prove this we have made use 
of radioactive carbon (Cis). This was produced 
by bombarding boron oxide in the cyclotron. Bi- 
carbonate containing this radioactive carbon was 
then prepared and injected intravenously into 
dogs. The distribution of this radioactive bicar- 
bonate between the pancreatic juice and blood 
plasma could then be followed by measuring the 
radioactivit)' of samples of these fluids collected 
at various intervals after the injection. Now if 
all the bicarbonate of the juice is derived from the 
plasma, then we may expect to find the radioac- 
tive liicarbonate concentrated in the juice to the 
same extent as the total bicarbonate. If no juice 
bicarbonate is derived from the serum, then there 
should be no radioactive material in the juice. If 
the juice bicarbonate is a mixture derived from 
both plasma bicarbonate and metabolic COo then 
the radioactive bicarbonate content of the juice 
should be in between these two extremes, and be 
a measure of the proportion of bicarbonate de- 
rived from the plasma. The results of four ex- 
l^eriments uniformly showed that the radioactive 
bicarbonate appeared promptly in the pancreatic 
juice in a concentration four to five times that 
found in the blood plasma. The total bicarbonate 
of the juice in these experiments was also four to 
five times that of the blood plasma. The results 
therefore indicate that the bicarbonate of the pan- 
creatic juice is largely derived from the blood 
stream. 

The pancreas is thus able to effect a four to five 
fold concentration of the plasma bicarbonate. This 



raises the question : where does this concentration 
of bicarbonate occur — on passage of bicarbonate 
from the plasma to the pancreatic cells or on pas- 
sage from the cells to the juice? Analysis of pan- 
creatic tissue for bicarbonate shows that the mil- 
limols of bicarbonate per Kg of HoO within the 
cells is 60% of that in the blood plasma (see 
Table I). Concentration of bicarbonate does not 
therefore occur on its passage from the blood 
stream to the cells, but on passage from the cells 
into the pancreatic ducts. 

Table I. 

ELECTROLYTE DISTRIBUTION BETWEEN BLOOD 
PLASMA, PANCREATIC TISSUE AND PANCRE.^TIC 
JUICE. 

All values expressed in terms of mM/Kg H2O. 





Blood 
Plasma 


Pancreati 
Tissue 


Pancreatic Juice 




Slow 
Secretion 


Rapid 
Secretion 


Na 


155. 




158. 


158. 


K 


5.4 




6.0 


6.0 


CI 


120. 


71. 


108. 


30. 


HCO;, 


28, 


18. 


34. 


120. 








pH = 7.65 


pH = 8.40 



An explanation for this concentration of bicar- 
bonate based upon the behavior of the chloride ion 
may be offered. As shown in Table I, the con- 
centration of chloride within the pancreatic cells 
is also about 60% of that in the blood plasma. The 
])ancreatic cell, unlike most tissue cells, would 
thus appear to be freely permeable to the chloride 
ion of the blood stream. The passage of chloride 
from the cells into the ducts would appear, how- 
ever, to be a slow process since only when juice 
is secreted slowly does its chloride content re- 
semble that of the blood plasma. As the rate of 
secretion increases, the chloride ion appears to be 
unable to diffuse fast enough to match against its 
share of sodium and potassium ions. The bicar- 
bonate ion which appears to be more readily dif- 
fusible, therefore, takes the place of the chloride 
ion in order to maintain electroneutrality. There 
thus results a pancreatic juice high in bicarbonate 
and consequently possessing an alkaline pH which 
is more suitable to the action of the proteolytic 
enzymes which it carries, 

(This work was performed with the collaboration 
of Drs. H. Tucker, B. Vennesland, and A. K. Solo- 
mon. This article is based upon a seminar report 
presented at the Marine Biological Laboratory on 
July 8.) 



July 12, 1941 ] 



THE COLLECTING NET 



57 




Bulbs for Scopelite (and Other Substage Lamps) 

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58 



THE COLLECTING NET 



[ Vol. XVI, No. 140 




EXHIBIT 

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its emphasis on fundamental prin- 
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July 12, 1941 ] 



THE COLLECTING NET 



59 




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60 



THE COLLECTING NET 



[ Vol. XVL No. 140 




Edward Bausch Microscope Maker 



WHILE Pasteur and his contemporaries were 
fighting the combined forces of superstition 
and disease to lay the foundations for modern 
bacteriology, another young man was designing 
a microscope that would help immeasurably in 
spreading the benefits of science to all mankind. 

While Pasteur was proving that heating would 
destroy the organisms that were making French 
wines turn bitter, and perfecting the pasteurizing 
process that makes his name immortal, in America, 
Edward Bausch was computing his own objectives, 
grinding his lenses and fitting the parts for the 
first Bausch & Lomb Microscope. 

While Pasteur was proving his procedure for the 
cure of rabies by saving the life of the little Alsatian 



peasant, Joseph Meister, Edward Bausch was 
working day and night to demonstrate his belief 
that quality microscopes could be made in quan- 
tities and at such prices as to bring them within 
the reach of all students and research workers. 

Today— you'll find Bausch & Lomb Micro- 
scopes in all the far corners of the world. Scientists 
in education, medicine and industry alike, know 
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than those bearing the Bausch & Lomb Trademark. 

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Vol. XVI, No. 4 



SATURDAY, JULY 19, 1941 



MAGNETIC MEASUREMENTS ON COM- 
POUNDS OF BIOLOGICAL SIGNIFICANCE 

Dr. L. Michaelis 

Member, Rockefeller Institute for Medical 

Research 

Dianiagnetism and Paramagnetism are analo- 
gous to induced electric dipoles and permanent 
electric dipoles. In magnetism, there is no phe- 
nomenon analogous to a free 
electric charge. A further dif- 
ference between magnetism 
electricity is the fact that elec- 
tric moments, whether induced 
or permanent, may vary wide- 
ly with respect to their magni- 
tude, whereas diamagnetic mo- 
ments are always very small, 
always involving repulsion by 
an external magnet pole ; and 
paramagnetic moments, if they 
exist at all, are always rela- 
tively large and cause attrac- 
tion by a magnet pole. All 
matter is diamagnetic, but in 
certain chemical compounds, 
paramagnetism is superim- 
posed on dianiagnetism, and 
being much larger, makes the 
latter almost negligible. It may 
be added that the phenomenon 



% % (iBlenisit 



TUESDAY, July 22, 8:00 P. M. 

Seminar: Dr. G. H. Parker: "The 
Melanophore System of Teleosts." 

Dr. R. L. Watterson: "Some As- 
pects of Pigment Deposition in 
Feather Germs of Chick Em- 
bryos." 

Dr. H. L. Hamilton: "The Influ- 
ence of Hormones on the Differ- 
entiation of Melanophores in 
Birds." 

Dr. Hermann Rahn: "The Distri- 
bution and Development of the 
Melanophore Hormone in the Pi- 
tuitary of the Chick." 



FRIDAY, July 25, 8:00 P. M. 
Lecture: Dr. Dorothy Wrinch : "The 

Native Protein." 



THE MAPPING OF EGGS 

A LECTURE TO THE EMBRYOLOGY CLASS 

Dr. E. G. Conklin 

Eiiieritiis Professor of Biology. 

Princeton University 

I feel like apologizing for appearing before an 
audience of people who are doing so much im- 
portant work in this day and age, because I have 
to report on things which were 
done long ago. For nearly 
fifty years past I have been 
asked to lecture before this 
class in embryology and al- 
most always I have spoken on 
essentially the same subject, 
though not, of course, in the 
same terms or under the same 
topic. 

I was once introduced as 
"the friend of the egg," and I 
am proud to claim that title. I 
have always maintained that 
the egg is after all one of the 
most marvelous things in the 
world and there is never an 
end to thinking about some of 
the mar\'els and wonders of 
development. 

I am therefore to give you 
something of a historical re- 



of ferromagnetism exhibited by a few metals, view of the backgrounds of modern embryology, 
such as iron or nickel, practically only in the free The marvel of development never grows old or 
metallic state, is never {Continued on page 72) stale. In a single night an ascidian egg will be- 



TABLE OF CONTENTS 



The Mapping of Eggs, Dr. E. G. Conklin 61 

Magnetic Measurements on Compounds of Bio- 
logical Significance, Dr. L. Michaelis 61 

Pathology and Immunity to Infection with 
Heterophyid Trematodes, Dr. H. W. Stunkard 65 

Factors in the Lunar Cycle which may Control 
Reproduction in the Atlantic Palolo, Dr. L. 
B. Clark 66 



Metabolism and Fertilization in the Starfish 

Egg, Dr. H. Shapiro 67 

Book Reviews 68 

Editorial Page 70 

Items of Interest 71 

Physiology Class Notes 73 

Botany Class Notes 73 

Embryology Class Notes 74 

Map of Woods Hole 75 




JEAN LOUIS RUDOLPHE AGASSIZ 



Who 68 years ago founded the Andersen School of Natural History 
on the Island of Penikese, which was the forerunner of the Marine 
Biological Laboratory. The bust was unveiled in the Hall of Fame 
in 1928 and is the work of Anna Vaughan Hyatt. 



July 19, 1941 ] 



THE COLLECTING NET 



63 



come a swimming tadpole. At six o'clock in the 
evening it may be fertilized ; at six o'clock in the 
morning it will be out of its egg membranes. In 
the case of Molgiila, it will be swimming actively 
only twelve hours after fertilization. 

In a few days an Arbacia egg is a swimming 
pluteus. In a month Crepidula becomes a fully 
formed, swimming veliger. In nine months the 
human egg becomes a baljy. While some of us 
labor for years to bring forth a brain-child in an 
article or book, nature brings forth a man-child 
in nine months. The brain-child is a fleeting 
thing, the man-child is a link in the endless chain 
of life. 

Development as we now understand it means 
both differentiation and integration. Differentia- 
tion is the becoming different of various parts. 
From the times of the ancient Greeks down to the 
year 1759 it was generally believed that this de- 
velopment consisted chiefly of growth of a previ- 
ously formed organism. Hippocrates said that 
blue-eyed parents produced blue-eyed germs. That 
was the belief that was held generally. Indeed, it 
was thought to be necessary philosophically, logic- 
ally and theologically. It was supposed that de- 
velopment must be the unfolding of an infolded 
organism and consequently it was not nearly so 
mysterious and wonderful a process as we now 
know development to be. 

In 1680 Antony van Leeuwenhoek, with simple 
lenses — a simple lens in a copper plate — studied 
the spermatozoa of man and different animals and 
described them quite accurately. Other people, 
taking his study as a basis, claimed that the sper- 
matozoa of man contained the homunculus, the 
little man. They reported it with head and body 
and arms and legs. 

I was reading only yesterday in our excellent 
library, some of Leeuwenhoek's letters in the ear- 
lier volumes of the Philosophical Transactions of 
the Royal Society. He wrote 125 letters which 
were published in some of the first twenty-four 
volumes, and in these letters he says that his ob- 
servations have been made sensational by other 
people. He knows perfectly well that there is no 
such homunculus in the spermatozoon. Further- 
more he says we know that in earlier stages of 
development the embryo is spherical while these 
sensationalists maintained that the arms were 
stretched out and the legs straight. Yet he did 
say that the man was pre-determined in the sperm 
and that in the uterus the sperm merely found a 
place to develop. With the prevalent masculine 



psychology of the time, the seed was supposed to 
come entirely from the male, while the female fur- 
nished a place, a location for its development. 

Well, this notion of the pre-determination of 
the man in the sperm was carried still further by 
van Leeuwenhoek in a letter which is not pub- 
lished in the Philosophical Transactions of the 
Royal Society, but which I came across in a 
Dutch publication discussing the work of van 
Leeuwenhoek. According to this, in 1680 he 
wrote and published the statement that the sperm 
are of two kinds, one male-producing and the 
other female-producing. When I showed this 
statement to Professor Wilson in 1905 or 1906, 
following his great discovery, he said, "Verily 
there is nothing new under the sun." This dis- 
covery of male-producing and female-producing 
spermatozoa was not actually made but was anti- 
cipated by Leeuwenhoek in 1680. Again those 
that were making a sensation out of his discover- 
ies said that Leeuwenhoek had discovered boys 
and girls in the uterus. That was nonsense, he 
said, but there are male-producing and female- 
producing sperm. He was getting at the idea of 
development by differentiation as contrasted with 
pre-formation. 

However, it was not until the middle of the 
eighteenth century that this doctrine of preforma- 
tion was carried to such absurd lengths that it 
simply broke down of its own weight. The Gene- 
van naturalist. Bonnet, was largely responsible for 
this. You will find a very fine account of Bon- 
net's work by Professor Whitman, in the "Lec- 
tures from the Woods Hole Laboratory," for 
1896, I believe. Professor Whitman reviews the 
work of Bonnet and points out the beautiful work 
that he did but the very erroneous conclusions to 
which he came. The conclusions were that there 
was no development. The embryo was infolded 
and it merely unfolded. Between the halves of a 
jjeanut, you can see the little leaves and stem and 
root of a plant, and you will see how easy it was 
to believe that in the earliest stages there was the 
little plant or the little animal in the seed. And 
this conclusion might have stood even longer than 
it did, for in addition to Bonnet, it had the sup- 
]5ort of many others including the physiologist 
Haller, who said "There is no becoming", that is, 
the plant or animal has been there from the start. 
They reasoned that if there was a little homuncu- 
lus in the sperm or in the egg, of course the little 
homunculus had sex organs and germ cells, and 
inside those there was the next generation, and 



The Collecting Net was entered as second-class matter July 11, 1935, at the Post Office at Woods Hole, Mass., 
under the Act of March 3, 1879, and was re-entered on July 23, 1938. It is devoted to the scientific work at 
marine biological laboratories. It is published weekly for ten weeks between July 1 and September 15 from Woods 
Hole, and is printed at The Darwin Press, New Bedford, Mass. Its editorial offices are situated in Woods Hole, 
Mass. Single copies, 30c by mail ; subscription, $2.00. 



64 



THE COLLECTING NET 



inside those the next, and so the doctrine of em- 
botUnent or encasement arose, and became so ab- 
surd that it had to be abandoned. 

Then a young medical student, Caspar Fred- 
erick Wolff, published his doctoral thesis in 1789, 
entitled "Theoria Generationis". He made an 
actual study of the hen's tgg and of certain plant 
seeds and found that in the earlier stages there 
was no little animal or plant in the tgg, but he 
found instead what he called globules. These 
globules we now call cells. He found membranes 
of some gelatinous material but no trace of an or- 
ganism, no trace of a living thing. He considered 
that the development was a process of differentia- 
tion caused by forces in nature to which he gave 
the name nisus formafhus or vis directrix. These 
forces from the outside, acting upon these glo- 
bules or membranes brought about development, 
and therefore since they were from the outside 
the whole process was called epigenesis. The 
word epigenesis meant genesis upon the germ, 
from outside stimuli. 

For a hundred years or more after this discov- 
ery of Wolff, the doctrine of epigenesis was uni- 
versally accepted and was held to be absolutely 
true. In the lectures of Professor Whitman you 
will find his answer to the assertions of Profes- 
sor Bourne of Oxford, who maintained absolute 
epigenesis, namely that there is no organization 
in the egg, and development comes from the out- 
side. \\^hitman showed the absurdity of this. He 
asked how is organization going to be "cooked 
into the egg" which lacks it? \\niat iiisus forma- 
tivus, what vis directrix is he going to demand to 
bring into the unorganized egg organization and 
capacity for development? 

I have made a translation from a book which 
is in the library and which I wish you would see. 
It was by one of the eminent zoologists of a pre- 
vious day, Alexander Goette. In 1874 he wrote 
a very large treatise, together with a volume of 
plates, entitled "Entwicklungsgeschichte der 
Unke," the "Development of the Toad," from 
v\'hich I will quote parts of pages 35 and 77. This 
is what Goette wrote in 1874 : 

The egg of Bombinator igneus ready for fertiliza- 
tion is neither in whole nor in part, neither in origin 
nor in its developed appearance a cell, a living or- 
ganism, but is merely an essentially homogeneous 
mass enclosed in an externally formed membrane, 
which first begins, through fertilization, to produce 
in itself living forms. But before this end is reached, 
the entire yolk mass and individual yolk pieces are 
lifeless transition stages from unorganized material 
to an actual organism. 

Just think of that, as late as 1874 the egg was 
not a living thing ! 

In 1898, when Jacques Loeb announced his dis- 
covery here at this Laboratory of artificial par- 
thenogenesis, the newspapers immediately took it 



[ Vol. XVI, No. 141 



up. "Jacques Loeb had created life! By putting 
an egg in a solution of magnesium chloride, he 
had created life." But of course the egg was just 
as certainly alive as is the animal into which it 
develops, though not as completely alive, for there 
are degrees of living. 

In 1883 another distinguished physiologist, 
Pfliiger, maintained that the egg of the frog is 
isotropic, that is in every direction from the cen- 
ter there are exactly the same substances and its 
cleavage cells are indifferent ; they are all exactly 
alike with "no more relation of the cells to the 
adult body than the snowflakes have to the result- 
ing avalanche." The snowflakes are not prede- 
termined to form an avalanche and the cells of the 
egg are not differentiated for the building of the 
body of the animal. 

In 1893 Hans Driesch, whose death last April 
has saddened all of his good friends in this coun- 
try announced a similar conclusion. We knew 
him and admired him although in many respects 
his work was wrong, I had my difficulties with 
him in the matter of the ascidian egg. When I 
found that parts of the ascidian egg gave rise to 
only a partial larva, while Driesch maintained that 
such larvae were entire, I said that the difficulty 
was that Driesch did not recognize the difference 
between a half larva and a quarter one ; if only 
these larvae had legs, Driesch would have seen 
that they were not complete. Driesch replied, 
"Conklin does not think well of experimentalists, 
but at least this much we know — the difference 
between a quarter and a half." 

His doctrine was like that of Pfliiger, that the 
egg was isotropic and all the blastomeres were 
equivalent, having no specific relation to develop- 
ing organs. He said that every blastomere of the 
sea urchin egg is equipotential, i.e., it is capable of 
giving rise to the entire organism. "Jedes kann 
Jcdes." Every cell can give rise to the whole or- 
ganism, and again he said that blastomeres were 
"like balls in a pile", so that you can shift them 
around any way you please, and still they would 
develop normally. In order to account for normal 
development, he had to invent something like a 
vis directrix. Something of that sort was neces- 
sary to make equipotential cells differentiate into 
a normal organism ; something had to intervene, 
presumably from the outside, and direct the de- 
velopment, and that something Driesch called, as 
you know, cntelechy. The entelechy of Driesch 
has many resemblances to the vis directrix of 
Wolff, or the "perfecting principle" of Aristotle. 
While I have argued against this in certain writ- 
ings in the past, maintaining that names of mys- 
terious forces that you can't experiment with are 
no solution of a problem — Driesch said you can't 
experiment with entelechy — as I get older I be- 
gin to see the necessity of something in living 



July 19, 1941 ] 



THE COLLECTING NET 



65 



things that has not yet been explained by rigid 
mechanism. I don't doubt that sometime it will 
be explained, that sometime we will find that mat- 
ter and energy are so constituted that they may 
account for all vital phenomena. I am greatly 
impressed with the fact that living things experi- 
ence satisfactions and dissatisfactions. We know 
that we have satisfactions and dissatisfactions, 
and there are evidences that higher and lower ani- 
mals also experience these. Even Paramecia. 
swimming in a trough of water which is hot at 
one end and cold at the other avoid extremes of 
heat and cold. Why? We would say because 
the water is getting uncomfortable. There is 
something subjective, internal in Paramecia that 
causes them to draw back from things that are 



uncomfortable. That may be called a tropism, or 
a negative reaction to extremes in temperature. 
\\'hy do they have this negative reaction? There 
is something in the living substance that feels, 
something that corresponds to the subjective phe- 
nomena that we have. Don't you see, I am com- 
ing around a little to sympathize with Driesch and 
his cntelechy. The future of biology will have to 
deal with these phenomena which we call subjec- 
tive feelings — satisfactions. Maybe their causes 
can be found in chemical and physical phenom- 
ena, such as "affinity," saturation, equilibrium, 
etc. Well, you see, I am getting 'way off into 
speculative regions. 

(Couchided in Next Issue) 



PATHOLOGY AND IMMUNITY TO INFECTION WITH HETEROPHYID 

TREMATODES 

Dr. Horace W. Stunkard 
Professor of Biology, New York University 



The term heterophyid refers to a large family 
of digenetic trematodes which infect fish-eating 
birds and mammals. Hcterophyes hcterophycs, 
the type species, was discovered by Bilharz 
(1851) in autopsies in Cairo. These trematodes 
are world wide in distribution and manifest not- 
able lack of specificity in their final hosts. Every 
species so far tested will develop in the common 
laboratory animals ; many of them have been re- 
corded from human hosts and probably all are 
possible parasites of man. Members of the fam- 
ily have lophocercous, oculate, monostome cer- 
cariae which encyst in fishes and so eventually 
reach their final hosts. Looss (1899) first point- 
ed out that the infective larvae must be acquired 
by eating uncooked fish. 

Yokogawa (1913) found a high incidence of 
infection with Metagoniunis yokogawai in Japan- 
ese, and, following the dictum of Looss, discov- 
ered the metacercariae in food fishes of Japan. He 
traced the development of this species in experi- 
mental animals and by human autopsy. Yokogawa 
reported that young worms, after liberation from 
their cysts, penetrate the wall of the intestine 
where they develop to sexual maturity, after 
which they return to the lumen of the intestine 
and eggs of the parasite are voided with the feces. 
Essentially similar findings in experiments with 
related species were reported by Ciurea (1924) in 
Roumania and by Faust and Nishigori (1926) in 
the Far East. Ciurea emphasized the possibility 
of secondary infection of lesions produced in the 
intestinal wall by migration of the developing 
worms. 

Africa and his collaborators, in the Philippines, 
found intramucosal invasion by various hetero- 
phyid species, but, according to these authors, the 



worms which enter the tissues do not return to 
the lumen of the intestine. Instead, they remain 
in the intestinal wall where they destroy the 
glands and erode the tunica propria but elicit only 
a mild or no inflammatory reaction. As a conse- 
quence, they are not walled off by fibrosis and 
their eggs pass into the general circulation. The 
eggs localize at various places, especially in the 
walls and valves of the heart. Autopsy of many 
cases of acute cardiac failure revealed extensive 
involvement of the heart, and these lesions were 
regarded as the immediate cause of death. Ex- 
perimental infection of various animals yielded 
divergent results and led Africa and his associates 
to the opinion that when introduced into natural 
hosts, the young worms remain in the intestine 
and produce a transitory and relatively harmless 
infection, whereas when they are liberated in the 
intestine of unusual hosts, they find the condition.^ 
unsuitable and bore into the intestinal wall. 

Cryptocotyle lingua is a common heterophyid 
species in the Woods Hole area, where it nor- 
mally infects gulls, terns and various mammals. 
Its life cycle was reported by Stunkard (1930) 
who attempted to infect different experimental 
animals. The larval stages are produced in Lit- 
torina Uttorea. and L. rudis, while the cercariae 
encyst in the cunner and other fishes. 

Stunkard and Willey (1929) studied the de- 
velopment of C. lingua in cats and rats. In these 
hosts, the worms developed to sexual maturity 
between the intestinal villi and no intramucosal 
invasion was observed. Since there is evidence to 
indicate that cats and rats are not favorable hosts, 
the studies were continued using terns and dogs. 
Results of these experiments, done jointly with 
Dr. C. H. Willey, were illustrated by lantern 



66 



THE COLLECTING NET 



[ Vol. XVI, No. 141 



slides. Young terns, removed from their nests 
soon after hatching, were kept in the laboratory 
and maintained on cunners heavily infected with 
cysts of C. lingua. They developed a very severe 
infection from the 6th to the 14th day, when the 
number of eggs in the feces began to diminish. 
After the 20th day the feces contained very few 
eggs and large numbers of young worms recently 
liberated from their cysts. The results of the ex- 
periment supplement and clarify observations on 
infection in nature. Young gulls and terns, yet 
unable to fly, are always heavily infected while 
adult birds, although subject to continual rein- 
fection or if kept in confinement and fed thou- 
sands of cysts each day, seldom harbor more than 
15 to 20 mature worms. It is apparent that after 
an initial heavy infection, gulls and terns develop 
a strong resistance to superinfection and the pres- 
ence of a few worms serves to maintain a substan- 
tial immunity. In terns, the young worms, when 
they emerge from their cysts, penetrate between 
the villi and disappear. Casual e.xamination of 
the surface shows few i)arasites but sections of 
the intestine show that the worms, although they 
invade deeply between the villi and cause much 
desquamation, rarely if ever enter the tissues. 
After the first heavy infection, when resistance 
develops and almost all of the parasites are shed, 
the intestinal epithelium is regenerated and the 
bird does not suffer any apparent ill effect. 

The experiments with dogs yielded very differ- 
ent results. A dog. fed enormous numbers of 
cysts, began to pass eggs of the parasite on the 
5th day. It was killed at this time and the intes- 
tine examined. Large numbers of immature and 
mature worms were present on the surface of the 
mucosa and in the crypts between the villotis folds. 
The villi showed acute inflammatory changes, 
desquamation, hyperemia and excessive mucous 



secretion. There was no invasion of the intestinal 
glands or tunica propria. Another dog, similarly 
fed for 14 days, was in a moribund condition. 
Killed at that time, autopsy revealed the presence 
of thousands of sexually mature worms. Sections 
of the intestine showed a copious exudate, which 
in places almost filled the lumen. The exudate 
contained strands of tissue, pieces of sloughed 
epithelium, and numbers of worms. Many worms 
were present also between the villi, deep in the 
crypts of the mucosa. In places, they had pro- 
duced pressure atrophy, marked hyperemia of ad- 
jacent villi, extensive desquamation and destruc- 
tion of the villi. There was some leucocytic in- 
filtration, accumulation of eosinophils and plasma 
cells, with regenerative hyperplasia of the intact 
epithelium. The erosion of the mucosa resulted 
in an acute catarrhal enteritis with proliferation 
of fixed tissue elements in the adjacent areas. No 
I)arasites were found in the tunica propria or in 
normal intestinal glands. Dogs were given a mod- 
erate infection and allowed to recover. Eggs be- 
gan to appear in the feces on the 5th day, were 
numerous for about 4 weeks, after which the 
number began to decline. At the end of 3 months 
\-ery few eggs could be found and the feces were 
negative at the end of 6 months. After resistance 
haci been established in dogs, the feeding of large 
numbers of metacercariae produced no visible ill 
eff"ects and \ery few eggs appeared in the feces. 

These experiments show that birds and dogs, 
if the latter survive an initial infection, effect a 
"self-cure" (as that term was defined by Stoll, 
1929) and thereafter are resistant to any substan- 
tial reinfection. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on 
July 15.) 



FACTORS IN THE LUNAR CYCLE WHICH MAY CONTROL REPRODUCTION IN 

THE ATLANTIC PALOLO 

Dr. Leonard B. Clark 

Assistant Professor of Biology, Union College 



Of all the physiological influences attributed to 
the lunar cycle, the coincidence of reproduction of 
certain marine Polychaetes with specific phases of 
the moon has been best determined. Of such ani- 
mals, the palolo worms are perhaps outstanding 
l)ecause of their size, their striking reproductive 
behavior and the apparent specific relation be- 
tween the moon's phases and time of reproduc- 
tion. 

The only palolo in the Western Hemisphere is 
the Atlantic palolo, Leodice fucata. found in 
Bermuda, the Gulf of Mexico and the West 
Indies. It is a burrowing form with the gonads 
limited to the segments of the posterior half of 



the worm except the last score or so. At repro- 
duction the sexual segments break from the rest 
of the body and swim to the surface where they 
discharge their sexual products. There is gen- 
erally typical mass activity so that many investi- 
gators concluded that the reproductive period is 
concentrated into a few days during which the 
sexual ends appear at the surface in immense 
numbers. 

The work of Mayer, and Clark and Hess shows 
that at least three factors serve to determine the 
particular day on which the majority of worms 
reproduce. Water turbulence induced by wind 
action will inhibit swarming if the wind velocity 



July 19, 1941 ] 

exceeds nine miles per hour unless the reef in- 
habited by the worms is protected. 

Records of swarming, 1898 to 1939, show that 
reproduction occurs most commonly about the 
time of the third quarter moon, less commonly at 
the first quarter, seldom at the full moon and 
never at the new moon, during the period from 
June 28 to August 1. This indicates that in some 
way the third quarter moon is most effective, fol- 
lowed by the first quarter, full moon and new 
moon. 

Worms will not reproduce at the third quarter 
moon less than 353 days from the date of repro- 
duction of the previous year nor less than 369 
days at the first quarter. The maturity of the 
worms determines the earliest date for reproduc- 
tion. The difference in time of maturity at which 
swarming will occur at the third and first quar- 
ter moon phases is correlative evidence that the 
third quarter moon phase is more effective than 
the first quarter in inducing reproduction. 

Hempelmann believed that the effect of the 
lunar cycle was indirect on Nereis dumerlii. He 
considered that tides changing with the moon 
caused changing nutritive conditions which 
brought Nereis into phase with the lunar cycle. 
Friedlaender suggested that changes in hydrosta- 
tic pressure due to tides controlled reproduction 
in the Pacific palolo, Eunice viridis. Neither of 
these hypotheses will hold for the Atlantic palolo. 
Mayer showed that animals in open floating cars 
reproduced at the normal time. Obviously there 
was no change in hydrostatic pressure. The 
change in nutritive conditions in transferring 
rocks containing worms from their natural habitat 
to live cars must have changed the nutritive con- 
ditions of the worms much more greatly than any 
change in tides. Also, rocks containing worms 
were scrubbed clean of the organic debris on 
which the worms feed, placed in floating cars and 
the worms swarmed normally. 

Furthermore, shading the worms in live cars 
from moonlight for more than two days imme- 
diately before the time of swarming inhibits rep- 



THE COLLECTING NET 



67 



roduction. These observations, first made by 
Mayer, were repeated recently in two successive 
years with similar results. 

At the present state of our knowledge, it seems 
almost impossible to escape the conclusion that 
moonlight acts directly on the Atlantic palolo to 
determine the time of reproduction. 

A number of experiments on artificially chang- 
ing the light relations of the lunar cycle by illum- 
inating or shading racks containing worms were 
undertaken. The results of all the experiments 
are consistent in that if the average duration of 
light is increased, reproduction occurs before the 
controls, and if the average duration of moonlight 
is decreased, the time of swarming occurs after 
the controls or not at all. It is concluded, there- 
fore, that this is a factor involved in reproduction 
and the effectiveness of the various phases of the 
moon's cycle is correlated with the average dura- 
tion of moonlight during the cycle. 

However, if this were the only factor involved, 
the effectiveness of moonlight to induce swarming 
would increase to a maximum about three days 
after the full moon and then decrease. But the 
effectiveness of moonlight is bimodal. the modes 
centering about the first and last quarter moon, 
with the latter much more effective. Obviously 
there must be some other factor operating in 
moonlight. The only other factor varying in the 
desired manner is the daily difference in the rate 
of change of moonlight. This reaches a maxi- 
mum at the new and full moons and a minimum 
at the first and third quarter. If it is postulated 
that the effectiveness of moonlight in determining 
the time of swarming bears some correlation to 
the average duration of moonlight during the 
Ivmar cycle and bears some correlation to the 
reciprocal of the difference in the daily rate of 
change of moonlight, the resultant varies in a 
manner similar to the incidence of swarming 
during the lunar cycle. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on 
July 15.) 



METABOLISM AND FERTILIZATION IN THE STARFISH EGG 

Dr. Herbert Shapiro 
Department of Physiology, Vassar College 



In his book "Artificial Parthenogenesis and 
Fertilization" (1913), Loeb summarized his 
views on the nature of fertilization by stating that 
"one essential effect of the entrance of the sper- 
matozoon into the egg of the sea-urchin is the 
acceleration of processes of oxidation". Although 
this is a good generalization for the sea urchin 
egg, it is not valid as a generalization for all eggs. 
Whitaker demonstrated that in the eggs of the 
clam Cumingia, (J. Gen. Physiol., 15, 183, 1931) 



and the worm Chaetopteriis, (J. Gen. Physiol.,-Z(5, 
475, 1933) there is not a rise, but a fall of respir- 
atory activity on fertilization, and in making a 
survey of all available data, tried to abstract some 
common element. On plotting out the absolute 
rates of oxygen consumption of different eggs be- 
fore and after fertilization, he observed (J. Gen. 
Physiol., 16. 497, 1933) that while the rates were 
quite divergent before fertilization, they became 
much more nearly the same after fertilization, and 



68 



THE COLLECTING NET 



[ Vol. XVI, No. 141 



proposed that the alteration in rate at fertilization 
is such as to bring it down or up to a more or 
less common level, comparable with that of nor- 
mally growing cells. 

This thesis has in turn had to be modified 
owing to the discovery of Rubenstein and Gerard 
(J. Gen. Physiol., 17, 677, 1934) that the value 
of the ratio, fertilized rate/unfertilized rate 
(F/U) is not a constant in the egg of the sea 
urchin, Arbacia puiictulata, but depends upon the 
temperature, being highest at low temperatures. 
The significance of these findings is that the res- 
piration-temperature dependence function must be 
different in fertilized and unfertilized eggs, and 
comes about through an alteration in the nature 
of the active enzyme systems responsible for over- 
all oxygen uptake, which takes place on fertiliza- 
tion. 

Earlier measurements (by Loeb and Wasteneys, 
( Arch. Entwicklungsmechn. Organ,. 35, 555, 
1912) and by Tang (Biol. Bull., 61. 468, 1931) 
of the egg of the starfish Asterias forbesii, at a 
single temperature, had led to the conclusion that 
in this egg, there is no change in oxidative rate 
on fertilization. However, if one e.xamines the 
original data, it is noted that relatively low per- 
centages of clea\-age were obtained and that the 
respiratory changes were in both directions. It 
seemed advisable therefore to extend these obser- 
vations through measurements of eggs from a 
large number of individuals, and over a wide 
range of temperatures, Warburg respirometers 
were used, and batches of eggs which showed 
usually 90% or more development were obtained 
during May and Jime, which is the optimal sea- 
son for starfish eggs. The respiration of unfer- 
tilized eggs may be constant for periods of over 
ten hours, whereas that of fertilized eggs is rela- 
tively constant at first, and then exhibits a grad- 



ually increasing rate as embryological develop- 
ment progresses. Measurements were made at 
about thirty different temperatures, between 11.5 
and 27.8° C, and an average increase on fertiliza- 
tion of approximately 30 to 50% was obtained. 
The average value of F/U was not constant over 
the range investigated, but showed a slight decline 
with elevation of temperature. In view of these 
data we should include Asterias with other eggs 
showing metabolic change on fertilization, and not 
classify it as an exception. 

On examination of the F/U-temperature curves 
for the eggs in which these data are available viz., 
Sabcllaria alvcolata, Chactoptenis variopedatus, 
Arbacia punctulata and Asterias forbesii. it ap- 
]iears that these cells show curves which indicate 
that the temperature dependence of the activity 
of the respiratory enzymes of unfertilized eggs is 
different from that of fertilized in all cases, and 
that hence there is an alteration in the nature of 
the o.xidative enzyme system on fertilization. For 
Arbacia punctulata. this has been further demon- 
strated by \arious investigators by treating the 
eggs with agents known to affect these enzymes. 
In the other eggs, this conclusion is an inference 
from the temperature analysis, and further chem- 
ical studies are desirable. 

By itself, therefore, the absolute direction and 
amount of change is probably not of primary im- 
])ortance, but it appears to be rather the nature of 
the enzyme system. The metabolic aspect of 
Loeb's generalization may be retained if we modi- 
fy his original idea by stating that one of the es- 
sential features of activation is an alteration of 
the nature of the enzyme systems. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on 
July 8.) 



BOOK REVIEWS 



Fisheries Along Our Coast 



NEW ENGLAND'S FISHING INDUSTRY. Edward 
A. Ackerman. Illustrated, pp. 294. $4.00. Univer- 
sity of Chicago. 

The great diversity of New England's fisheries 
can scarcely be appreciated until one reads Mr. 
Ackerman's account of this industry. In a straight- 
forward easy to read style he has covered virtual- 
ly every phase of the field, both at sea and on 
land. His thorough research covers not only the 
major aspects of the fishing industry but includes 
man}' details seldom presented and which add 
considerable interest and value to his work. To 
the layman many of these factors are little known, 
and even for those persons whose business is fish- 



ing, or the handling of fishery products, there is 
a rich store of information. 

About one-half of the book deals directly with 
the various species of fish, mollusks and crusta- 
ceans, that are caught off the New England coast 
and landed in its numerous ports. Seasonal 
abundance, migrations, and value of catchall are 
adequately described as are fishing grounds, fish- 
ermen, gear, methods of capture, and the limita- 
tions and regulations that govern the fisheries. 
The description of the various types of gear used 
for catching fish, clams, quahogs, oysters, scal- 
lops, squid, crabs, lobsters, etc., is of especial in- 
terest. In the case of fish, the relative efficiency 
and seasonal productivity is given for weirs, 
pound nets, otter trawls, gill nets, trawl lines, 



July 19, 1941 ] 



THE COLLECTING NET 



69 



hand lines, and other devices. An account is 
given of the kinds of fish taken by such kind of 
gear. 

The remaining chapters of the book give an 
account of the marketing and handling of fish and 
fishery by-products and of the economic aspects 
of the industry. They explain, for example, how 
the more perishable or the less desirable species 
can not compete in distant markets but how, nev- 
ertheless, fresh fishery products are penetrating 
farther afield because of improved transportation 
facilities and methods of handling. Also described 
is the role of the salt fish trade and the reasons 
for its great decline during the past few decades. 

The author has included many illustrations, an 
index, and after each chapter a list of the more 
important references. There are likewise many 
tables giving the amounts and value of the New 
England catch in recent years, both domestic 
catches and importations, and in some cases these 
are segregated according to the methods of cap- 
ture that were employed. Various points dis- 
cussed are illustrated on a series of thirty-three 
charts. They include the distribution and magni- 
tude of the catch of the more important species, 
fishing grounds, polluted areas along shore, loca- 
tion of weirs, and the location of canneries. 

W. C. Schroedcr 



A Standard Text 



WOODRUFF, L. L. "Foundations of Biology." pp. 
xvii -|- 773. New York, The Macmillan Company. 
1941. $3.75. 

New editions of classic texts are difficult to re- 
view impartially. The appearance of six editions 
in less than twenty years is clear evidence of in- 
trinsic worth so that enthusiastic praise descends 
from the level of critical judgment to a politically 
expedient endorsement of public opinion. Ad- 
verse criticism is even more unfortunate for it can 
only be justified by laboriously selecting those 
slight blemishes which can l^e found in any work 
and then endeavoring to maintain the case that 
they are major faults. The soundest test is prob- 
al)Iy to inquire whether the vohune still justifies 
its title. 

Biology is a vast subject defined in the first 
]iaragraph of the book under review as "the 
science of matter in the living state". How far, 
then, does the volume continue adequately to de- 



scribe the foundations of this science? This de- 
pends entirely on what the reader regards as the 
foundations of liiology. Woodruff's view is very 
clear for in Chapter II "we now turn directly to 
the study of life itself in the only form it is known 
— the bodies of plants and animals". This leads 
to a discussion of the cell and its development 
into a multicellular organism, followed by three 
chapters on plants and four on animals. Then 
follow five rather conventional chapters on the 
comparative anatomy of organ systems leading to 
two regretably short, but thoroughly enjoyable, 
chapters on endocrine and nervous coordination 
which serve to link the previous descriptive biol- 
ogy to the subsequent more philosophical discus- 
sions. These start with the origin and continuity 
of life as an introduction to fertilization, develop- 
ment and genetics, pass logically to adaptation 
and the origin of species and the book ends with 
the humanistic and historical aspects of the 
science. 

All this, of course, is superbly well done. 
Woodruff has evidently read everything which is 
worth reading and possesses the happy knack, 
shared by the honey bee, of re-presenting what 
he has digested in a form more palatable than it 
was before. In this he has been aided and abetted 
by an excellent artist (Miss Lisbeth Krause) and 
a publisher who has devoted unusual skill and in- 
telligence to the design and production of the 
book. Criticism, however, still remains possible. 
If we accept "the bodies of plants and animals", 
i.e. comparative anatomy, as the true foimdation 
of the study of life we are struck by the anomaly 
that less than one third of the volume is devoted 
to this study and that the remaining two thirds, 
while making pleasant reading, do not inflict a 
very rigid mental discipline on the student. No 
one knows better than Woodruff that the basis of 
all science is logic and that mathematics is the 
language of logic. Yet nowhere in the non-de- 
scriptive portions of the book does one find any 
warning to the freshman that, if he is successfully 
to continue his biological studies, he will be con- 
strained to do so in a thermodynamical frame- 
work as rigid as that which surrounds even the 
freshman chemist or physicist. An introduction 
to the mathematics of growth, and an indication 
that it is possible for a species to have statistical 
validity, might well have been included among 
the "Foundations of Biology". 

There is no doubt, however, that the sixth edi- 
tion will retain the high rating among general 
biology texts which has already been gained by 
the previous five. Peter Gray 



70 



THE COLLECTING NET 



[ Vol. XVI, No. 141 



The Collecting Net 

A weekly publication devoted to the scientific work 
at marine biological laboratories. 

Edited by Ware Cattell with the assistance of 
Boris I. Gorokhoff and Judy Woodring. 

Entered as second-class matter, July 11, 1935, at 
the U. S. Post Office at Woods Hole, Massachusetts, 
under the Act of March 3, 1879, and re-entered, 
July 23, 1938. 



M.B.L. CLUB 

The Annual Meeting of the M.B.L. Club will 
be held at the Chibhouse on July 21, 1941, at 
7:00 P. M. 

Order of business : 
Minutes of last annual meeting. 
Reports of officers. 
Reports of committees. 
Election of officers for the next year. 
Consideration of proposed amendment to constitution. 
General business. 

All members are urged to attend this meeting. 

Proposed Amendment to the Constitution: Ar- 
ticle X shall be amended to read : "A building 
fund shall be maintained to be used for the re- 
pair of, or improvements to the Clubhouse. In 
this fund the Treasurer shall deposit each year 
10% of the dues collected that year. The fund 
shall be administered by the Trustees and money 
withdrawn from it only by their action on request 
from the Executive Committee of the Club. Fur- 
ther, in accordance with an agreement with the 
Marine Biological Laboratory, the Treasurer shall 
pay to the Laboratory 25 % of the dues collected 
each year." 

At present Article X provides that 25% of an- 
nual dues be put into a building fund. In fact 
this payment has been tised in repaying the lab- 
oratory for repairs and improvements made sev- 
eral years ago. A year ago the Club came into 
an agreement with the Laboratory whereby the 
Club is to regularly make a payment of 25% of 
its annual income from dues and the Laboratory 
will take care of the maintenance of the grounds, 
exterior, and structure of the building. This pay- 
ment represents a payment of rent to the owners 
of the building. The Club will continue to be 
responsible for the interior of the Clubhouse. The 
second part of the amendment proposed will ratify 
this agreement. 

The first ])art of the amendment as proposed 
will establish a new building fund that may be 
used for major repairs or improvements inside 
the Clubhouse, or for further enlargements. 

Nominations for next year's officers will be pre- 
sented at the meeting by the Nominating Com- 
mittee, consisting of Drs. H. B. Goodrich, Samuel 
E. Hill, T. H. Bullock and J. Htitchens. 

— Sears Crozvell 



ADDITIONAL INVESTIGATORS 

Beck, L. V. instr. phys. Hahnemann, lib. 

Boche, R. D. instr. zool. Pennsylvania. Br 221. D 
112-A. 

Bronfenbrenner, J. prof. bact. & immun. Washing- 
ton Med. (St. Louis). Br 305. 

Garner, H. res. asst. zool. Chicago. Br 332. Ka 22. 

Goldinger, J. M. res. asst. med. Chicago. Br 125. K 
14. 

Gurewich, V. Cornell Med. lib. 

Kopac, M. J. asst. prof. biol. New York. Br 311. A 
106. 

Kreezer, G. L. asst. prof, psych. Cornell, lib. D 312. 

Mead, F. W. Ohio State. Br 111. 

Metz, C. W. prof. zool. Pennsylvania. Br 304. 

Morgan, Isabel M. asst. path. & bact. Rockefeller 
Inst. (New York). Br 320. 

Reiner, J. M. biophysics (New York, N. Y.). lib. 

Renshaw, B. asst. prof. zool. Oberlin. Br 218. 

Sturtevant, A. H. prof. biol. California Tech. Br 126. 

Warren, A. A. asst. path. Harvard Med. L 27. 

A NOTE FROM DR. DURYEE 

The following paragraphs are extracts from a 
letter received by Dr. Robert Chambers from Dr. 
William R. Duryee, assistant professor of biol- 
ogy at Washington Square College, New York 
University. Dr. Duryee was a reserve officer and 
is serving as lieutenant in the U. S. Army. 

Your film was delivered to me in my truck just 
as my column was about to roll out on the road. We 
have been moving so often from one bivouac to the 
next that there has been no time to acknowledge 
your kindness in sending the film. 

At present we are moving the 27th Division back 
from Tennessee to Alabama. What with 28 new 
2y2 ton 6X6 "ten-wheelers" and new 1% ton and 
command cars along with the old, our regiment 
looks quite respectable out on the road — about three 
and a half miles long. As transportation officer I 
am charged with shuttling our Regiment and several 
battalions of Infantry. We go day and night for 
four days. It is good fun for me but pretty hard 
on the drivers who alternate 2 hours sleep and two 
driving. 

Now that our first big maneuvers are over, with 
all the dust, "jiggers" (by the way what is the biol- 
ogy of these mites ? ) and lack of water, there will be 
quite a let-down. Most of the boys will get ten-day 
furloughs. Our Colonel has emphasized the need 
for interesting lectures, films, training of any sort 
for this in-between period. Yoiu" film is therefore 
exceptionally welcome. Can you send three more 
16 mm ? Capillaries, microdissection, mitosis — any 
and all are needed. 

Can it be that Woods Hole and the M. B. L. still 
exist? I feel rather remote from cell physiology; 
but two days ago I found some lizard eggs in an 
old log and dissected them for some of my company. 
The lizards were well-developed with large yolk sacs 
attached and motility had set in. I wish I had these 
while teaching embryology. 



CURRENTS IN THE HOLE 

Date A. M. P. M. 

July 19 12:39 12:50 

July 20 1:30 1:43 

July 21 2:20 2:31 

July 22 3:05 3:15 

July 23 3:49 4:00 

July 24 4:30 4:43 



July 19, 1941 ] 



THE COLLECTING NET 



ITEMS OF INTEREST 



Dr. and Mrs. Charles Packard will be at 
home to members of the Laboratory tomorrow 
and next Sunday from four-thirty to six o'clock. 

Dr. Curt Stern has been promoted from as- 
sociate professor of zoology at the University of 
Rochester to professor of experimental zoology. 

Dr. Carl G. Hartman, of the department of 
embryology of the Carnegie Institution of Wash- 
ington at Johns Hopkins University, has been ap- 
pointed professor of zoology and head of the de- 
partments of zoology and physiology at the Uni- 
versity of Illinois beginning September 1. 

Dr. John B. Buck, instructor in zoology at 
the University of Rochester, has been promoted to 
an assistant professorship. 

Dr. E. E. Reinke, head of the Department of 
Biology at Vanderbilt University, has been named 
Chairman of the Division of Natural Sciences of 
the University. 

Dr. H. H. Plough presented a lecture at eight 
o'clock on Thursday evening to the class on em- 
bryology. He spoke on "Genes and Develop- 
ment." 

Dr. Caswell Grave will be the guest lecturer 
before the embryology class on Tuesday, speak- 
ing on "Ascidian Metamorphosis." 

Guest speakers before the physiology class next 
week will include Dr. Samuel E. Hill, who will 
speak Monday on "The Action Current of Nitel- 
la," and Dr. E. S. Guzman Barron, who will lec- 
ture Tuesday on "Oxidation-Reduction Systems 
in Cellular Respiration." 

Dr. Wm. Randolph Taylor will give a talk 
at the botany seminar on Thursday evening en- 
titled "The 1934 Allan Hancock Expedition." 

Dr. Victor Jolles died in Madison, Wiscon- 
sin, on July 5. He was formerly on the staff of 
the Kaiser Wilhelm Institute in Berlin, and later 
for two years (19,37-1938) at the University of 
Wisconsin. He is well known for his work on 
Daucrmodifikationen in Protozoa and later for 
work on heat-induced mutations in Drosophila. 

The 1941 Conference on Spectroscopy and its 
Applications will be held at the George Eastman 
Research Laboratories at the Massachusetts In- 
stitute of Technology from July 21 to July 23. 

Among the exhibits in progress at Woods Hole 
this week are those of the Bausch and Lomb Op- 
tical Company at the Coast Guard Canteen, the 
Clay-Adams Company at the Old Lecture Hall, 
and the Macmillan Company in the Lobby of the 
Brick Building. 



Miss Eleanor Blevins is being married today 
to Dr. Donald J. Zinn, who has worked at the 
Lalioratory in past years, at St. Barnabas Church 
in Falmouth. 

Miss Dorothy Wellington, who is doing re- 
search work at the Laboratory, was married on 
June first to Dr. K. L. Osterud at her home in 
Melrose, Massachusetts. Dr. Osterud, who is 
from Ashton, Virginia, received his doctor's de- 
gree this past June from New York University, 
is working at the Marine Biological Laboratory 
this summer, and this coming academic year will 
teach at the University of Minnesota. 

Dr. Detlev W. Bronk, director of the John- 
son Foundation for Medical Physics at the Uni- 
versity of Pennsylvania, and Dr. H. S. Gasser, 
Director of the Rockefeller Institute for Medical 
Research, spent last weekend at Woods Hole. 

Dr. Paul A. Weiss, associate professor of 
zoology at the University of Chicago, visited 
Woods Hole on July 11 and 12. He is teaching 
at the University this summer. 

Dr. Leonor Michaelis left Woods Hole with 
his family for northern New England yesterday. 

Miss Louise "E. Boyden, editorial assistant on 
the Biological Bulletin, returned to Woods Hole 
this week after a two-week trip to Maine where 
she visited Mrs. H. B. Neal at the Mt. Desert 
Biological Laboratory and afterwards stayed at 
Penobscott Bay. 

At the Dartmouth Conference, the following 
men were elected to the executive committee of 
the Society for the Study and the Development 
of Growth: Dr. Paul A. Weiss, chairman; Dr. K. 
V. Thimann, secretary ; Dr. J. W. Wilson, treas- 
urer; Dr. B. H. Willier, Dr. O. L. Sponsler and 
Dr. E. W. Sinnott. 

The program for the weekly phonograph record 
concert at the M.B.L. Club next Monday is as 
follows : Hindemith, "Trauermusik" ; Shostako- 
vich, "Symphony No. 1"; intermission; Wagner, 
"Parsifal — Prelude and Good Friday Music"; 
Wagner, selections from "Gotterdammerung." 

A "poverty dance" will be held at the M.B.L. 
Club tonight, at which dancers will wear rag cos- 
tumes. 

The United States War Department has placed 
an order for 52,000 laboratory mice with the 
Jackson Laboratory at Bar Harbor, Maine. 
These mice will be used for medical purposes in 
the national defence program. 



72 



THE COLLECTING NET 



[ Vol. XVI, No. 141 



ITEMS OF INTEREST 



The ketch Atlantis of the Woods Hole Oceano- 
graphic Institution sailed on July 10 for a cruise 
to the continental shelf just south of Georges 
Banks to take borings of the ocean bottom under 
the direction of Dr. Henry T. Stetson, research 
associate in paleontology at the Harvard Museum 
of Comparative Anatomy. Dr. Theodore White. 
Fred Phleger and \\'alter Hamilton make up the 
remainder of the scientific party. The ship is ex- 
pected to return about July 20. Early this week 
the power boat Anton Dohni sailed for a routine 
trip under the supervision of Mr. F. Fuglister. 



Dr. and Mrs. Thomas P. Hughes are spend- 
ing the month of July in Woods Hole on their 
30-foot motor cruiser anchored in the Eel Pond. 
Dr. Hughes is a member of the field staff of the 
International Health Division of the Rockefeller 
Foundation and for the past two and one-half 
years has been studying the epidemiology of yel- 
low fever in the Uganda Protectorate, Africa. He 
and Mrs. Hughes will return to Uganda for fur- 
ther work if war conditions permit, and plan to 
sail from New York on August 20. 



MAGNETIC MEASUREMENTS ON COMPOUNDS OF BIOLOGICAL SIGNIFICANCE 

(Continued from page 61) 



encountered in chemical compounds of biological 
interest. 

All chemical compounds with all their valences 
saturated, and containing only atoms with com- 
pleted electronic shells, are diamagnetic. There- 
fore the overwhelming majority of organic com- 
pounds is diamagnetic. The main interest lies in 
paramagnetic molecules. They arise under two 
conditions. The first case is represented by com- 
pounds containing atoms with uncompleted elec- 
tronic shells. Of these, iron is most important 
for the biologist. The second case is represented 
by chemical compounds, especially organic ones, 
such as possess an odd number of electrons, in 
other words, compounds with a free, not occupied 
valence, usually designated as free radicals. All 
organic substances that can be reversibly oxidized 
and reduced, develop such a free radical as an in- 
termediate step of oxidation-reduction. This very 
property produces the reversibility of oxidation- 
reduction. The existence of such free radicals in 
reversible dyestuft's such as methylene blue, pyo- 
cyanine, the yellow respiration ferment, has been 
previously demonstrated by a potentiometric 
method, sur\eyed last year by the author in one 
of the Friday-evening lectures in Woods Hole. 
The detection of paramagnetism during the pro- 
cess of reduction of such dyestuffs is another 
method to prove the existence of free radicals. 
This phenomenon is demonstrated by the lecturer 
in some lantern slides showing the appearance, 
and, later on, the disappearance, of paramagnet- 
ism during the process of reduction. 

Among the iron-containing compounds it is es- 
sentially hemoglobin and its derivatives which 
may show paramagnetism. The magnitude of the 
magnetic dipole moment can be utilized to eluci- 
date the chemical structure, especially the nature 
of the chemical bond between the iron atom and 
the surrounding complex-forming groups. Paul- 



ing and Coryell discovered that hemoglobin is 
strongly ]3aramagnetic, whereas oxyhemoglobin is 
not paramagnetic at all but shows only the slight 
diamagnetism characteristic of all non-paramag- 
netic substances. The speaker extended this 
study to crystallized catalase from beef liver. The 
method had to be refined and a differential micro- 
method has been developed, which was in princi- 
ple demonstrated in lantern slides. This method 
showed that catalase is strongly paramagnetic, yet 
not quite as strongly as hemoglobin, and corre- 
sponding more to that of alkaline methemoglobin 
such as found by Pauling and Coryell. 

The cause of paratnagnetism in free radicals is 
the spin of the odd electron, which is equivalent 
to a circular electric current. In ordinary valence- 
saturated molecules, the electrons are arranged in 
pairs, each consisting of two electrons with oppo- 
site spin, cancelling the magnetic effect of the spin. 
In molecules containing iron, the paramagnetism 
is due to the fact that there is an uncompleted 
electronic shell. In this case it is not necessarily 
true that the electrons are arranged in pairs with 
opposite spins. Some of the electrons may pos- 
sess spins in equal direction. In this case, the 
paramagnetic effect of an electron is not cancelled 
but even increased, each of the electrons with 
parallel spins contributing its share. In other 
cases, the electrons may be paired. In oxyhemo- 
globin, all electrons are paired ; hemoglobin con- 
tains five unpaired electrons with parallel spins. 
Catalase has four unpaired electrons. One should 
remember that the magnetic effect itself may not 
be of much significance, but the knowledge gained 
from the magnetic measurement in regard to the 
nature of the chemical bond will probably be of 
great value for the analysis of the chemical 
structure of such compounds. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on 
July 8.) 



July 19, 1941 ] 



THE COLLECTING NET 



7i 



PHYSIOLOGY CLASS NOTES 



This week Dr. Ballentine, the last member of 
the staff to deliver lectures, spoke on "Ultra- 
Micro Analyses for Biological Investigation," 
"Recent Advances in Histo- and Cyto-Chemis- 
try," and "Enzymes as Analytical Tools." Dr. 
M. E. Krahl was the guest lecturer on Thursday, 
his subject being "Interactions of Biologically 
Significant Substances in Surface Films." 

Dr. Ballentine is reported to be suffering from 
writer's cramp due to his failure to use free-arm 
movement while filling two blackboards with ref- 
erences. Tuesday's lecture was enlivened by a 
rare demonstration of what almost amounted to 
sky-writing. The janitors having considerately 
sponged away his carefully written blackboard 
lecture supplements, Dr. Ballentine with admir- 
able aplomb mounted the front table and replaced 
them amid the cheers of the three members of the 
class who were awake by that time. 

After a week of spirited competition in the high 
breakage derby, your correspondent and partner 
were successful in carrying off the top honors. 
Mr. and Mrs. J. Broken Van Slyke (the J. stand- 
ing for Just) are now at home to visitors in 
Room 10. 

Second prize was awarded to the Liinulus blood 
group for their successful studies of disintegra- 
tion processes in the Haldane-Henderson appara- 
tus. Individual honors were given for special 
effort to Herb Weiner by the contest judge, Mr. 
Ino Hoodonit. 



This week's issue finds the class excused from 
formal instruction and free to pursue the bypaths 
of individual research. At this time next week 
we expect to be able to report all sorts of dis- 
coveries. 

This week's issue also finds Dr. Fisher learn- 
ing to use the slide rule so that he can calculate 
in a hurry the results he got. 

The Physiology High Command in a recent 
communique announces the loss of two runs in a 
crucial engagement in the Park Street sector, in 
which a valiant stand was taken against the crew 
division by Gunner Bob Harrison and Field Mar- 
shals Ed Burns, Dr. Kempton, Dick Henry, Jim 
(Ersatz) Green, Dr. Kenneth (Panzer) Fisher, 
et al. An impressive mass demonstration was 
staged by a crowd of non-combatants. In other 
words, we had a very excellent cheering section 
with banners and everything. 

The hidden talent of one of our class members, 
Pat Perkins, has finally come to light in the con- 
tribution of the following Ode to Arbacia : — 

The life of a Physie 
Ever so busy 
Is incomplete 
Sans Arbacia neat. 
We praise ya, 
Arbacia. 

—Mr. and Mrs. J. B. V. S. 



BOTANY CLASS NOTES 



The halfway mark is passed ; five weeks are 
behind us, and only one is left. What has this 
time been to the botany students? Weeks spent 
investigating the Clorophyceae — the green algae, 
in which chlorophyll is the dominant pigment — 
and the Phaeophyceae — the browns, in which the 
chlorophyll is masked by the accessory pigments 
carotin, xanthrophyll, and fucoxanthin. Now the 
class is studying the third big group, the Rhodo- 
phyceae — red algae, most colorful because of the 
superabundance of phycoerythrin. 

Studying the algae involves 8 :30 lectures every 
morning by either Dr. Taylor or Dr. Runk. The 
rest of the day, to say nothing of the wee small 
hours of the night, is spent in lab where Mr. Gil- 
bert and Mr. Tseng are kept busy running from 
table to table, assisting in the identification of 
species which seem so similar to the untrained 
students. Each collecting trip is bound to reap 
several of the innumerable species of Entero- 
morpha and Polysiphonia, and many a student 
has been disappointed to find that his cherished 



mount of Enteromorpha miniina is nothing but 
E. intestinalis which he found on the first trip and 
has mounted every time since. Dr. Taylor, how- 
ever, is still optimistically hoping that at least a 
quorum will cease trying to elevate the weed, 
Ceramium ruhrmn to a rare genus before the end 
of the course. 

A week ago Wednesday, the collecting trip was 
to Penikese Island, and, as usual, the Nereis left 
Eel Pond at 8:30 a.m., with the lazy members of 
the class just making it. Luck with good weather 
held, so that the trip to the island and the walk 
around it were most enjoyable. Due to a high 
wind the day before, there was a rich wash of 
"seaweed" along the shore, with the result that 
many species not previously found were picked 
up in sorting out the drying algae. Only three 
pieces of the feathery Plumaria plmnosa were 
found, however, which made the lucky finders 
prey to the ravenous sink-collecting scavengers 
that night in lab. Another rarity was the beauti- 
ful autumnal oak-leaved Phycodrys. All truly 



74 



THE COLLECTING NET 



[ Vol. XVI, No. 141 



spirited collectors had a great time getting thor- 
oughly soaked in grabbing for algae in the quiet 
second between breakers on the windward side 
of Penikese. Once a leper colony and formerly 
the original M.B.L. station, the island is inhabi- 
ted now by only gulls and terns, whose nests are 
so numerous that to cover the island without 
stepping on the protectively colored eggs was a 
real feat. 

Dr. F. B. Smith, agronomist and professor at 
the University of Florida, had charge of the Bot- 
any seminar last Thursday night. His lecture was 
on the microorganisms found in the Florida soils. 
In addition to generally telling about the countless 
and varied inhabitants of earth, he brought out 
the fact that their presence largely determines the 
quality of the soil ; hence, Florida ground, which 
has little life per square inch as compared with 
that further north, is quite poor for cultivation. 
After the seminar, his wife served delicious date 
cake and whipped cream — there went the figures ! 
— with real Chinese tea, prepared by Mr. Tseng. 

All-day picnics, it seems, with boats for trans- 
portation are apparently the vogue in this neck 
of the woods, Init since the Botanists had already 
visited most of the island spots — via boat — on 
field trips, they made their class picnic last Tues- 
day a late afternoon and all-evening affair to the 



sand dunes near Sandwich. Upon arrival, every- 
one went either swimming or sliding down dunes 
in search of wood, then spent two and a half 
hours singing around the bonfire after a huge 
meal. Sufficient indication that the affair was a 
success was the remark made by Dr. Taylor, a 
confirmed picnic-hater, "It wasn't half as bad as 
I expected it to be !" 

Getting back at twelve p.m., however, had no 
noticeable effect on anyone but Father Stenger 
who failed to put in appearance by 8 :30 the next 
morning when the Nereis set out for a collecting 
trip to Gay Head. As is the customary technique, 
the three skiff-loads of algologists waded through 
the wash around the Head for several hours be- 
fore returning to the boat for lunch. Collecting, 
nevertheless, was not too profitable — except to 
slow-pokes who still had neither Laminaria nor 
Cystoclonhmi — and though the shore waters were 
laden with algae, few new things were picked up. 
Later homeward bound, the Nereis stopped in 
two places to make dredgings, but, unfortunately 
for the vegetarians, except for a little Phyllo- 
phora broadaei, the total haul included only star- 
fish, bryozoans, shells, pebbles, and hermit crabs. 
In spite of this, and since it's always fair weather 
when the botanists are together, no one minded 
much and the trip was set down as an even better 
one than last week's. — /. W. and C. S. 



EMBRYOLOGY CLASS NOTES 



.Icadeuiic Life : 

On Tuesday, July 8, Dr. Schotte completed his 
career at Woods Hole in a blaze of echinodermal 
glory when he discussed differentiation of chem- 
ical material within the egg as an example of the 
influence which the SO4 ion has in the polarity 
of the tgg. We are sorry that this course will 
no longer have the privilege of Dr. Schotte's in- 
spiring lectures. This vivacious individual is 
planning to settle down to hard research (with 
no distractions) in the hills of Massachusetts. 

Dr. Costello opened the discussion on Annelids 
with a detailed description of the future of each 
blastomere. We were all a little relieved when 
the A^aiipliiis was finally produced with its three 
pairs of appendages and a simple eye spot instead 
of the complicated cell system which preceded. 

On Thursday we were on the receiving end of 
a classical lecture by Dr. Hamburger which con- 
cerned primarily his own work in the field of ex- 
perimental Neuroembryology. 

Friday came cell lineage and our minds were 
spun with spiral cleavage. Without Ray's beau- 
tifully stained slides of Crepidula cell stages we 
never would have been unwound. 

On Saturday, we had the pleasure of hearing 



Dr. Conklin, who gave an introduction on the 
argument of Preformative vs. Epigenetic Devel- 
opment and concluded with a brilliant discussion 
on "The IVIapping of Eggs." This lecture was 
one of the outstanding features of the course and 
a memorable experience for us all to have had 
the opportunity of hearing the man who fathered 
the School of American Experimental Embryol- 
ogy. The "Friend of the Egg" amused us all 
with his account of the "cell lineagists" and the 
"egg shakers" at this Laboratory. 

Monday, under the guidance of Dr. Hambur- 
ger, the Trochophore larva was studied with an 
air of semiaccuracy midst a cloud of subdued pro- 
fanity. The little blighter twister and turned and 
managed to conceal all vital structures beneath 
yolk and array of pigment. 
Social Life : 

The high spot occurred Tuesday when one of 
the beautiful days, highly valued because of their 
rarity, called forth a group of truants who, though 
they departed through the back door of the lab- 
oratory were unable to escape the vigilance of 
Dr. Costello. With face wreathed in smiles the 
good doctor bade them farewell with a "bon voy- 
age" and a mild suggestion tMit the laboratory 
be continued at sunset. — P. and K. 



July 19, 1941 ] 



THE COLLECTING NET 




76 



THE COLLECTING NET 



[ Vol. XVI, No. 141 




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July 19, 1941 ] 



THE COLLECTING NET 



77 




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Other ADAMS CENTRIFUGES and laboratory 
supplies are described in cur new Catalog No. 
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July 14th to Setli. 



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78 THE COLLECTING 


NET [ Vol. XVI, No. 141 


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July 19, 1941 ] 



THE COLLECTING NET 



79 



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4) 



THE COLLECTING NET 



[ Vol. XVI, No. 141 




The Strange Case of The Invisible Evidence 



■p\EATH had struck in the night. 

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Vol. XVI, No. 5 



SATURDAY, JULY 26, 1941 



Annual Subscription. $2.00 
Single Copies, 30 Cents. 



THE ORGANIZATION OF THE MELANO- 
PHORE SYSTEM IN BONY FISHES 

Dr. George How.vrd Parker 

Professor of Zoology, Emeritus 

Harvard University 

The common eastern catfish, Aiiieiiirns nebiilo- 
siis, can vary in color from pale yellowish green 
to coal black. This color change, which depends 
upon the melanophores in the 
skin of this fish, is excited 
through at least three recep- 
tors, the dorsal retina, the ven- 
tral retina, and the skin. The 
color changes in Ameiurus 
take place slowly and require 
for their completion more than 
a day. Consequently this fish 
is especially favorable for color 
work. 

The pale phase of the cat- 
fish is dependent upon the dor- 
sal retina and is best seen 
when the creature is over a 
white background illuminated 
from aljove. Under such cir- 
cumstances that light which 
passes directly from its source 
to the fish's eye enters that 
organ somewhat obliquely and, 
after passing across its inter- 
ior, impinges upon the ventral retina. Of the 
light that falls on the white background below the 
fish and is reflected (Continued on page 89) 



M. % Iff. €alcnftav 



TUESDAY, July 29, 8:00 P. M. 

Seminar: Dr. T. C. Evans and Mr. 
J. C. Slaughter: "Radiosensitivity 
of Arbacia Sperm under Differ- 
ent Conditions." 

Dr. J. P. O'Brien: "Effect of X- 
Radiation on Regeneration in 
Nais paraguayensis." 

Dr. A. M. Chase: "Effect of Azide 
on Cypridina Luciferin." 



FRIDAY, August 1, 8:00 P. M. 
Lecture: Dr. Ernst Mayr, "Specia 
tion in Birds." 



STORAGE AND DISTRIBUTION OF MAN- 
GANESE IN OSTREA VIRGINICA 

Dr. Paul S. Galtsoff 
U. S. Fishery Biological Laboratory 
Woods Hole, Mass. 

The tissues of marine Lamellibranchs store 
relatively large quantities of heavy metals. Fe, 
Cu, Mn, Zn and sometimes Pb were found in 

their bodies and it has been 

established that there is con- 
siderable variation in the first 
two of these elements in the 
oysters taken from various lo- 
calities along the Atlantic and 
Gulf coasts. Copper was found 
to be particularly abundant in 
the oysters from the northern 
states whereas those from the 
South Atlantic and Florida 
waters are richer in iron. 

The existence of definite 
seasonal changes in the glyco- 
gen and water contents of oys- 
ters and clams suggested that 
there may be similar seasonal 
fluctuations in the contents of 
heavy metals of these mol- 
lusks. To avoid variations 
which might have been attri- 
buted to racial differences, age, 
or location, oysters of known age and origin 
were planted on the experimental bottom in Long 
Island Sound near Milford, Connecticut. The 



TABLE OF CONTENTS 



Storage and Distribution of Manganese in 
Ostrea Virginica, Dr. Paul Galtsoff 81 

The Organization of the Melanophore System 
in Bony Fishes, Dr. G. H. Parker 81 

The Mapping of Eggs, Dr. E. G. Conklin 83 

The Rectifying Property of the Giant Axon of 
the Squid, Drs. R. Guttman and K. S. Cole.. 86 



The Distribution and Development of the Me- 
lanophore Hormone in the Pituitary of the 

Chick, Dr. H. Rahn ." 87 

Editorial Page 90 

Items of Interest 91 

Physiology Class Notes 92 

Botany Class Notes 92 

Embryology Class Notes 93 

Supplementary Directory for 1941 94 



July 26. 1941 ] 



THE COLLECTING NET 



83 



3000 Inishels of oysters planted on this area rep- 
resented a homogenous population from which for 
a period of aljout 3 years random samples were 
taken at bi-weekly intervals. Each sample con- 
sisted of ten oysters which were opened, weighed 
and placed in an oven for drying within three 
hours after they were taken out of water. 

Employing the procedure recommended by 
Richards (The Analyst, 1930, 55:554) the sam- 
ples were analyzed for Mn and other metals. The 
results revealed great regularity in seasonal varia- 
tions of the manganese content of these samples. 
The lowest concentration of Mn (between 7 and 
11 mg. p.k.d.w.) occurred during cold months 
( November- April ) whereas the highest concen- 
trations (between 35 and 55 mg. ]3.k.d.w.) coin- 
cided with warm seasons (May- August). A 
study of the sexual cycle of the oyster shows that 
during the latter period gonads undergo rapid 
development and the mollusks reach sexual ma- 
turity. Following the discharge of sex cells the 
Mn curve rapidly drops to its minimum. The 
regularity of the Mn-curve suggests that accumu- 
lation of this metal is somehow associated with 
the sexual phase of the oyster. This view is cor- 
roborated by the analysis of separated tissues of 
the oyster. For this determination samples each 
comprising 10 or 20 female or male oysters were 
prepared by excising the mantle, gills, gonad, 
muscle and visceral mass. The latter part always 
contained a small amount of gonad tissues which 
could not be completely removed. It has been 
found that ovaries had highest Mn content vary- 



ing from 51 to 59.6 mg. p.k.d.w. On the other 
hand the testes were very poor in Mn, having 
only from 4.6 to 7.i mg. p.k.d.w. In the gills the 
Mn varied from 17 to 18 mg. p.k. in winter and 
from 35 to 38.6 in summer. In the visceral mass 
the Mn varied from i'.9 to 18.4 mg. p.k. and in 
the adductor muscle its content in July was 4.3 
to 5.2 mg. p.k. and 4.1 to 9.3 in January. The 
mantle had a Mn- content of 8.7 mg. in January 
and from 14.2 to 17.0 mg. p.k.d.w. in September. 

Since the ovarv is obviously the principal organ 
in which the Mil is stored, it is reasonable to con- 
clude that high conteTit of this metal in mixed 
samples is primarily due to the presence of ripe 
females. The Mn cycle in Ostrea virginica is ob- 
viously associated with the development of the fe- 
male phase. The exact role of this metal in the 
metabolism of this mollusk and its possible effect 
on sex change is not known. It is reasonable to 
expect, however, that being a strong catalyst, its 
presence in the ripe female has physiological sig- 
nificance which we hope will be elucidated by our 
further experiments. 

Large quantities of Mn were found by Bradley 
in fresh water mussels (/. B. Ch. 1907 and 1910, 
V. 3 and 8) and Dubuisson et Van Heuverswyn 
(Arch. d. Biol., 1930, 41) have shown that in 
' Anodonta cygnea it is principally stored in the 
internal demibranchs. Its physiological role in 
mussels is not known. 

(This article is based upon a seminar leport pre- 
sented at the Marine Biological Laboratory on 
July 15.) 



THE MAPPING OF EGGS 

Dr. E. G. Conklin 

Emeritus Professor of Biology, Princeton University 

(Continued from last issue) 



I have spoken about embryologists who believed 
in extreme epigenesis, under the influence of vis 
directrix or entelechy. But coincident with their 
work was that of others who found that the egg 
and its blastomeres were not isotropic and equi- 
potential. In 1874, the same year in which Alex- 
ander Goette published his work on the toad's 
egg, Wilhelm His studied the hen's egg and con- 
cluded that the surface of the blastoderm could be 
mapped, certain portions being destined to give 
rise to the head and others to other parts of the 
embryo, that is, that the egg could be mapped into 
Organbildende Keimbczirke, organ-building germ 
regions. 



In 1877 Professor Whitman, afterwards the 
first director of this Laboratory, published his 
doctoral thesis on the embryology of Clepsine. 
While it was a little crude as compared with some 
of our modern work, it was very carefully done. 
He found that individual blastomeres gave rise to 
particular parts of the embryo, and he said that 
while the embryo is not predelineated in the egg, 
all its parts are predetermined. 

In 1879 Carl RabI studied the embryology of 
Planorhis and found that individual cells gave rise 
to particular parts of the embryo. Edward van 
Beneden in 1884 did the same thing for the as- 
cidian egg, pointing out that the very first cleav- 



The Collecting Net was entered as second-class matter July 11, 193,5, at the Post Office at Woods Hole, Mass., 
under the Act of March 3, 1879, and was re-entered on July 23, 1938. It is devoted to the scientific work at 
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84 



THE COLLECTING NET 



[ Vol. XVI, No. 142 



age is in the plane of bilateral symmetry, one blas- 
tomere giving rise to the right half of the body 
and the other to the left. That was an important 
discovery, to find bilateral symmetry established 
at this early stage. 

In 1887, a yovmg French student. Louis Chab- 
ry, studied the ascidian egg, following the meth- 
ods of Driesch. He found that one of the first 
two blastonieres gave rise to half a larva. Driesch 
maintained that it was a whole larva, but his pic- 
tures of ascidian larvae were very crude and in- 
conclusive. 

In 1888 Wilhelm Rou.x stated that the frog's 
egg from the four-celled stage on is a "mosaic 
work", each of these iwrtions giving rise to a 
quarter of the adult. Professor Wilson, in a paper 
which he first gave as a lecture here at the Lab- 
oratory and published in 1893 under the title, 
" Amphioxiis and the Mosaic Theory of Develop- 
ment," criticized this theory. Some person thought 
that this was the theory of Moses, the author of 
Pentateuch. 

In 1892 Wilson published his "Cell Lineage of 
Nereis." one of the great classics of embryology. 
In 1890 and 1891 he did this research at this Lab- 
oratory. At the same time I was occupying the 
Johns Hopkins Table at the Fish Commission and 
was working on the Embryology of Crcpidida. 
but had never met Wilson and knew nothing of 
his work on Nereis. W'ilson heard of what I was 
doing and one Sunday morning he came over to 
see my drawings of Crepidiila and to show me his 
of Nereis. I shall never forget our excitement. 
He had found, as I had, that there were three 
groups of cells that came off from the macro- 
meres, that these gave rise to the whole of the 
ectoderm, and that the very next division of one 
of these macromeres gave rise to the mesoderm 
of the body. Here were two animals belonging 
to phyla as far apart as the annelids and the gas- 
tropods that gave rise to embryos in an exactly 
similar way. So we came to the conclusion that 
seemed necessary in those days, that there must 
be some phylogenetic relationship, that they came 
from common ancestors. If homologies are evi- 
dences of common descent, why should this not be 
true of cells as well as of organs? Thus arose at 
Woods Hole the cult of cell lineage, including be- 
sides Wilson and myself, Lillie, Mead, Holmes, 
Treadwell, Surface and a number of others who 
were finding individual blastomeres destined to 
give rise to particular parts of an animal. 

But at the same time this was going on, the 
cult of "egg-shakers" became prominent at Woods 
Hole. This was about the same time that Driesch 
had broken eggs apart and found that each l)las- 
tomere gave rise to an entire organism. Morgan 
was going about the laboratory shaking eggs in 
test-tubes. Morgan, Loeb. Child and others had 
all found, as Driesch did, that fragments gave 



rise to whole animals. So the question arose, 
who is telling the truth? 

Apparently what was true of annelids and gas- 
tropods was not true of echinoderms ; conse- 
quently I proposed the terms "determinate" and 
"indeterminate" cleavage for these two types. 
Later work show-s that this is not a very useful or 
accurate distinction. The fact is that all eggs are 
partly determinate : at least . we now know from 
the work of Horstadius and others that this is true 
of echinoderms. In the case of the determinate 
eggs, they also have a certain amount of power 
of regulation. Consequently the difference is one 
of degree. One is relatively more regulative and 
the other is relatively more fixed. I found, for 
example, that ascidian eggs are highly mosaic, in 
consequence very definitely determinate, but in 
the case of Amphioxns, one-half of the egg will 
give rise to a whole animal, of half the size but 
beautifully developed. But if Amphioxus blasto- 
meres are separated in the four-cell stage, right 
or left halves will give rise to a whole larva, but 
anterior or posterior halves will not ; the anterior 
half-embryo would have the notochord and the 
neural plate but not the mesoderm. In the two- 
cell stage, each half has all the substances that 
are found in the other half and it needs only to 
close up the injured side l\v regulation to give 
rise to a whole embryo. The fact is that we are 
here dealing with a phenomenon that is wide 
spread throughout the animal kingdom, namely, 
regeneration, and the beliefs of the egg-shakers 
in this early period that because you could get a 
whole larva out of half or a quarter of an tgg, 
that cells were therefore undifferentiated, had 
about as good a foundation as the view that be- 
cause you can get a whole Hydra or Planaria or 
Annelid from a part of an animal, that therefore 
these animals are undifferentiated. 

Driesch said that it vias impossible to conceive 
of any machine that could be broken up in the 
three dimensions of space and the fragments re- 
main capable of producing whole machines, and 
he claimed that the egg could be broken up and 
each part then grow into a whole animal ; there- 
fore he considered that the mechanistic theory of 
development was false. He did not sufficiently 
realize that living things are much more complex 
than a watch. In each cell there is a nucleus 
which is undifferentiated, which can in some cases 
remould the cytoplasm so as to bring about res- 
toration of missing parts, because there is here 
a machine inside the machine, there is the outer 
machine which is the c}i:oplasm and there is the 
inner machine which is the nucleus, and as long 
as the nucleus is not broken up we may get re- 
generation. 

Well, this all leads to the point toward which 
I have been aiming, namely that differentiation in 
development takes place in the cytoplasm. There 



July 26, 1941 ] 



THE COLLECTING NET 



85 



is no differentiation in the nucleus. In some way 
the nucleus does control differentiation in the cy- 
toplasm : we know enough about wliat is happen- 
ing in the egg to recognize some of the stejxs in 
the nuclear control of the cytoplasm. 

In the case of many of the eggs you have been 
studying in this course you have found that dur- 
ing mitosis the nuclear membrane disappears and 
there is left in the cell body the relatively small 
chromosomes and a large amount of granular 
achromatic material which came out of the nu- 
cleus. The chromosomes then swell up and form 
chromosomal vesicles, and as the swelling goes 
on they get larger and larger, filling up with ma- 
terial taken from the cytoplasm until they form 
the large resting nucleus within which new chro- 
mosomes appear. At each division a large amount 
of achromatic material is set free into the cell 
body. I have followed this achromatic nuclear 
material in Crepidula up to the twenty-four cell 
stage and it always goes to that part of each cell 
which is nearest to the polar bodies ; here it re- 
mains as granular material and ultimately cilia de- 
velop right over these granules. Thus we get dif- 
ferentiation from material which came from the 
nucleus and mixed with material in the cytoplasm. 
In this way differentiation occurs in the cyto- 
plasm as a result of this "diastole" (swelling) of 
the nucleus and its "systole", when it breaks and 
releases material into the cell body. A certain 
amount of this interchange between nucleus and 
cytoplasm must take place in the absence of mito- 
sis, but in mitosis you can see it most clearly. In 
this way differentiation in the cytoplasm comes 
under the control of the nucleus. 

The localization of these different cytoplasmic 
substances in different cells and in different parts 
of the embryo is always a moving and in some 
cases a striking phenomenon. Eggs in which 
movements are rapid are most favorable. I don't 
know whether the Styela eggs are as good this 
year as they sometimes are. If you get animals 
which have a lot of orange color in the body, you 
will find that the eggs will have a large amount of 
that orange pigment and then you can see the 
sorts of things which are shown in this chart. In 
the early stages the pigment is scattered over the 
surface of the egg. Immediately after fertiliza- 
tion, this surface layer flows down to the point 
where the spermatozoon has entered. This ma- 
terial containing the yellow pigment then flows to 
the posterior side of the egg where it forms a 
crescent which gives rise to mesoderm. Another 
crescent of light gray material around the anter- 
ior side of the egg gives rise to the notochord and 
the neural plate. Between these crescents on the 
ventral side is the material of the ectoderm, on 
the dorsal side that of the endoderm. A similar 
mapping of the egg is found in Amphioxus and 
Amphibia. Regulation in the case of isolated 



blastomeres of these eggs depends to a certain 
extent, at least, upon the fluidity of their cyto- 
plasm. 

The late Dr. E. P. Lyon in 1906 used a centri- 
fuge in .some physiological work which he was 
doing here, and he tried it on sea urchin eggs and 
found that he could stratify the substances of the 
egg. This was a new instrument for studying 
the substances of eggs. Morgan and Lyon in the 
following year tried this out on eggs of the sea 
urchin and Cuniingia, and they found that irre- 
spective of where the pigment and yolk went, the 
embryo was perfectly formed. The conclusion 
was that these egg substances were not morpho- 
genetic. 

I centrifuged ascidian eggs and found that I 
could throw the yellow pigment, which is the 
lightest substance in the egg. to almost any point 
and still get normal development, so that this pig- 
ment is not morphogenetic. The muscles which 
normally contain this pigment will form when it 
is lacking ; however when I centrifuged much 
harder, I found that I did dislocate materials that 
were morphogenetic, and then I could get animals 
with the muscles and chorda outside the body, 
with the ectoderm inside, with the eye spots inside 
or outside the body, indeed could bring about an 
abnormal distribution of the organs of the body 
by bringing about this abnormal distribution of 
morphogenetic substances in the unsegmented 
egg. When these substances of the ascidian egg 
are displaced by high centrifugal force, they pro- 
duce what Virgil in the Aeneid called mcmhra 
disjecta, organs scattered around in various ways 
out of position. I take this as evidence of the fact 
that we have morphogenetic substances in these 
eggs. No one has ever questioned the fact that 
the cytoplasm of the cells of the liver is different 
from that in the ganglion cells. The question is 
how early in the development can you find such 
differences, and I find that certain differences can 
be carried back to the one- or two-celled stage. 
Amphioxus eggs are just as plainly mapped-out 
as are those of ascidians There is here also a 
mesodermal crescent around the posterior half of 
the egg and a chorda-neural crescent around the 
anterior half, an endodermal area on the dorsal 
side and an ectodermal one on the ventral-anter- 
ior side. A similar mapping of the Amphibian 
egg is seen after fertilization in the "gray cres- 
cent" (chorda-neural) around the anterior side 
and a mesodermal crescent around the posterior. 
Yung has carried the mapping of these eggs much 
farther by locating on the egg the presumptive 
areas of all the principal organs of the embryo, 
but these areas are not generally marked out by 
visibly different kinds of ooplasm as is the case 
with the two crescents and the ectodermal and 
endodermal areas. 

In 1901 Boveri found that soon after the fer- 



86 



THE COLLECTING NET 



[ Vol. XVI, No. 142 



tilization of the egg of the sea-urchin, Strongylo- 
centrotus, a pigmented zone forms just below the 
equator. Below this pigmented zone is a clear 
area which gives rise to the four micromeres 
which form mesenchyme, the pigmented zone 
forms endoderm, while the area above the pig- 
ment becomes ectoderm. The three "germinal 
layers," ectoderm, endoderm, and mesoderm, are 
mapped out in this egg before cleavage begins. 

The localization of morphogenetic substances in 
Annelids and Molluscs differs from that in Echin- 
oderms, but in many respects resembles that in 
Amphioxus and Ascidians. The mesoderm comes 
chiefly from an area on the posterior side of the 
egg, while the ectoderm comes from the area 
around the animal pole and the endoderm from 
the vegetal portion of the egg. 

In some of the hydro-medusae (e.g.. Litierges) 
a concentric localization of substances in the egg 
is visible at an early stage, and in normal develop- 
ment the outer layer forms ectoderm, the inner, 
endoderm. 

In all phyla there is a polar differentiation of 
the egg before fertilization, the upper or animal 
pole, at which the polar bodies form, giving rise 
later to the ectoderm, while the endoderm usual- 
ly comes from materials of the opposite or vegetal 
pole. The axis connecting these poles bears a 
characteristic relation in different phyla to the 
chief a.xis of the developed animal. In some eggs 
bilateral symmetry or asymmetry is plainly vis- 
ible before cleavage. And in a few favorable eggs 
even the location of the materials of particular 
organ-systems such as nervous system and sense 
organ, muscles and mesenchyme, notochord and 
endoderm are mapped out on the egg in areas of 
different colors or textures. 

In conclusion, the egg is a living organism and 
its differentiations are the beginnings of the dif- 



ferentiations of the embryo and adult. Its polar- 
ity, symmetry and pattern give rise to those of the 
developed animal. This much of pre-formation 
of the future animal is present at the beginning of 
development. But the further details of develop- 
ment are the results of progressive differentiation 
resulting from the interaction of nucleus and cy- 
toplasm. The nucleus is the seat of the inheri- 
tance genes, the cytoplasm of all the differentia- 
tions of development. 

The differentiations of the cytoplasm begin at 
different stages and reach different degrees in dif- 
ferent species and classes of animals, but always 
some differention begins before the fertilization 
of the egg. Therefore, the egg contributes more 
to development than the sperm does. The differ- 
entiations of the spermatozoon, i.e., acrosome, 
middle piece, tail, are lost after it enters the egg, 
but the differentiations of the egg persist. The 
spermatozoon reaches the end of its development 
before it is ready to enter the egg. but the egg 
reaches the end of its development only with the 
fully-formed animal. An egg can develop into 
an adult without the stimulus or contribution of 
a sperm, but no one has ever been able to get 
spermatozoon to develop apart from an egg. We 
derive more from our mothers than from our 
fathers. We are vertebrates because our mothers 
were vertebrates and produced eggs of the verte- 
brate pattern, but the color of our skin and eyes 
and hair and other features of later development 
are determined by the chromosomes from the 
sperm as well as those from the egg. The egg 
only undergoes embryonic differentiation. 

I am still "the friend of the egg." 

(This article is based upon a lecture delivered 
before the Embryology Class at the Marine Biologi- 
cal Laboratory on July 12.) 



THE RECTIFYING PROPERTY OF THE GIANT AXON OF THE SQUID 



Drs. Rita Guttman and Kenneth S. Cole 
College of Physicians and Surgeons, Columbia University 

Law. Ohm's Law states that the current is pro- 
portional to the voltage in a conductor over a 
wide range of voltages and currents, and the fac- 
tor of proportionality may be termed the resis- 
tance, i.e. 



The permeability of a living membrane to ions 
may be expected to be proportional to its electri- 
cal conductivity, since both of these properties 
depend upon the ease with which ions may pass 
through the membrane. 

On passing an electric current through a living 
membrane, one finds that the electrical conductiv- 
ity is greater under the cathode and less under the 
anode than normal. It has been assumed that the 
ion permeability is correspondingly increased un- 
der the cathode and decreased under the anode. 

If the electrical conductivity or resistance (elec- 
trical conductivity is the reciprocal of electrical 
resistance ) thus varies under the electrodes, the 
living membrane cannot be said to follow Ohm's 



= R 



I 



where E represents electromotive force in volts, 
I represents current in amperes, and R is the re- 
sistance in ohms. 

According to Ohm's Law, then, the resistance 
of a conductor should be the same under the elec- 



July 26, 1941 ] 



THE COLLECTING NET 



87 



trades regardless of the magnitude of the cur- 
rent and regardless of the direction of the cur- 
rent but it has been found not to hold for the Hv- 
ing membrane of the squid axon. Since the mem- 
brane of the squid axon permits current to pass 
more easily in one direction (outward), than in 
the other (inward), it is a rectifier rather than a 
pure resistance. 

In two papers published in March, Cole, Baker 
and Curtis demonstrated rectification in the squid 
axon, using alternating current bridge methods 
and the needle electrode technique. These papers 
show that the previous failure of Cole and Hodg- 
kin to observe rectification in this axon was due 
to the facts that only small currents had been used 
and that the effect under the anode was approxi- 
mately equal and opposite to the effect under the 
cathode, thus neutralizing it. 

In the work here reported, which was done at 
Woods Hole last summer, rectification was ob- 
served in the squid axon directly by means of a 
much simpler technique involving a direct current 
Wheatstone bridge. One end of the fiber was im- 
mersed in sea water, the interelectrode stretch was 
in oil and the other end of the fiber was injured 
by dipping it into KCl, so that a current travelling 
through the fiber passed across only one mem- 
brane. Thus neutralization of rectification under 
one electrode by rectification under the other was 
avoided. 

The resistance of the fiber was measured dur- 
ing the passage of currents of varying magni- 
tudes. The direction of the current was reversed 
at every reading thus minimizing polarization of 
the fiber membrane. Currents were sent in for 
short times only, and considerable and equal time 
intervals elapsed between readings, so that even 
though currents considerably above rheobase were 
often used, the condition of the fiber did not 
change materially over long periods of time. Dur- 
ing a run there were frequent returns to a refer- 
ence EMF value so that any shifting of resistance 
values with time and with temperature changes 
could be compensated for in the calculations. 

The measuring technique used in these experi- 
ments has certain advantages. It is simple. It 
permits one to use an axon immediately after dis- 
section. On the other hand, runs take about 



twenty minutes. The immediate problem is to 
speed up the measurements. 

Simultaneous readings on excitability, resting 
potentials and rectification were made on single 
fibers as the fibers died. As the fiber dies not 
only does excitability disappear and the resting 
potential approach zero, but rectification, also, is 
gradually lost. The completely dead nerve fiber 
membrane acts like a pure resistance rather than 
a rectifier. 

Progressive and partially reversible loss of rec- 
tification also accompanies narcotization of the 
nerve fiber with cocaine or veratrine sulphate. 

The measurements show then that rectification 
is lost when the nerve fiber becomes inexcitable, 
when it dies, when it is anesthetized. Rectifica- 
tion thus seems to be associated somehow with 
the normal functioning of nerve. 

Impedance and membrane potential measure- 
ments indicate that the rectifier efifect is situated 
in the membrane, and it seems reasonable to as- 
sume that membrane conductance is a measure of 
ion permeability. These rectification measure- 
ments then indicate that there is an increased ion 
permeability under a cathode and a decreased ion 
permeability under an anode. Interpreting in a 
similar manner the results of Ebbecke obtained 
on frog nerve, we find large permeability changes 
at both anode and cathode, superimposed upon 
which there was a small permeability difference 
between anode and cathode regions. Blinks also 
studied rectification in the giant plant cell, 
Valonia. 

In conclusion, rectification may assist in ex- 
plaining various puzzling alternating current 
phenomena in nerve, such as change in membrane 
potential during the passage of an alternating 
current, the delay of excitation with alternating 
currents, alternating current block, alternating 
current break excitation, the time of rise of elec- 
trotonus being more rapid at the cathode than at 
the anode. However, an explanation of rectifica- 
tion, itself, is not as yet available. The explana- 
tion of rectification will probably also be an ex- 
planation of the mechanism of the ion permeabil- 
ity of the membrane. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on 
July 8.) 



THE DISTRIBUTION AND DEVELOPMENT OF THE MELANOPHORE HORMONE 
IN THE PITUITARY OF THE CHICK 



Dr. H. 

Instructor in Zoology, 

The intermedin hormone or the melanophore 
dispersing principle is commonly derived from the 
pars intermedia of the typical vertebrate pituitary 
gland. In the birds, however, this hormone pre- 
sents a peculiar problem, because a structural pars 
intermedia is absent. Furthermore, its effect 
upon the bird melanophore or any other function 



Rahn 

University of Wyoming 

it may have in the bird or mammal has not been 
demonstrated. However, this hormone is present 
in considerable quantities in the chicken pituitary 
gland as tested by its pigment dispersing qualities 
in lower vertebrates. It is therefore not to be re- 
garded as a hormone that has lost its function in 
the warm blooded animals, but very probably 



THE COLLECTING NET 



[ Vol. XVI, No. 142 



plays an entirely different role which still awaits 
discovery. Such a change in hormone function 
during the course of phylogenetic development 
must be kept in mind, since, for example, the pro- 
lactin hormone of the pituitary has been demon- 
strated to exert widely different effects in the 
various vertebrate groups. In amphibia, this hor- 
mone seems to induce the migration urge of the 
land newt to seek a water environment, in the 
bird it acts directly on the crop gland of the pig- 
eon, and in the mammal it is concerned with milk 
secretion in the mammary gland. 

The more immediate attack upon the melano- 
phore hormone problem as presented here may be 
outlined as follows : ( 1 ) where is this hormone 
found in the absence of a structural pars inter- 
media? and (2) does a study of the formation of 
this hormone during development give us any pos- 
sible clue to its function? 

Morphology of the bird pituitary. At the time 
this work was begun the presence or absence of 
a pars intermedia in the bird was still debated. A 
detailed cytological study of the development of 
the chicken pituitary gland revealed, however, that 
at no time during ontogeny or later on could such 
a structure be demonstrated. Furthermore there 
did not exist any connection between the infundi- 
bular process of the diencephalon and the glandu- 
lar portion, the pars buccalis. In the typical ver- 
tebrate there is an intimate connection between 
these two components contributed by the presence 
of a pars intermedia. Since the case of the chicken 
might possibly represent a specialized type (as 
has been found in some mammals which have no 
])ars intermedia) a comparative study was under- 
taken to see how widespread this morphological 
peculiarity is. Altogether 18 species were studied, 
representing 12 families and five orders. They all 
revealed a pituitary construction identical in pat- 
tern to the one found in the chicken. 

Distribution of the hormone. The infundibular 
process had for a long time been suspected in 
higher forms to yield at least part of the melano- 
phore hormone, since it was never possible to 
se]iarate it completely from the closely applied 
pars intermedia. In the chicken this separation 
is normally present and tests revealed that no 
melanophore hormone could be found in this 
nervous component. This then suggested that the 
])ars anterior is entirely responsible for hormone 
]M-oduction and it became of interest to find out 
just where it was found within the glandular com- 
ponent. From a phylogenetic standpoint one 
might expect it to reside in the region nearest the 
infundibular process. Quantitative assays, how- 
ever, revealed that it is found in all regions, but 
is 20 times more concentrated in the region fur- 
thest removed from the nervous lobe. 

After ascertaining the distribution of this hor- 
mone within the gland, the next problem con- 
cerned itself with the quantitative assay of the 



melanophore hormone during ontogeny. Prelim- 
inary tests and observations on amphibian larvae 
revealed that not only is hormone present in the 
pituitary, but also functioning in melanophore 
dispersion at an extremely early age. Histologi- 
cal sections revealed a very much undifferentiated 
pituitary gland. This rather unusual situation 
prompted a similar and more detailed investiga- 
tion in the chick pituitary, since in this form the 
cytological dift"erentiation had been worked out. 

Assay Method. For the quantitative determin- 
ation of the melanophore hormone the Anolis liz- 
ard was employed. This animal exhibits most 
striking color changes, from a bright green to a 
dark brown. The metachrosis is produced by the 
concentration and dispersion of pigment in the 
melanophores and is regulated by the melanophore 
hormone of the pituitary gland. When the pitui- 
tary complex is removed the animal is perma- 
nentlv green and becomes very sensitive to this 
hormone. In fact 1/10,000 of mg of dried bird 
pituitary tissue will evoke a darkening response. 

The degree of darkening is proportional to the 
hormone concentration injected into such a hypo- 
physectomized animal and four successive stages 
of darkening from bright green to dark brown 
have been assigned arbitrary values of stages 1-4. 
For the actual test a known fraction of the pitui- 
tary gland is injected into ten operated animals 
and their average color response recorded as 1,2, 
3, or 4. From several such determinations the 
exact concentration can be calculated which will 
evoke precisely a color response of stage 1. This 
concentration of hormone is then designated as 
1 J(nolis) [/(nit). {Anat. Rec, 76, 157). 

Hormone genesis. By this method the melano- 
phore hormone was determined for various age 
groups. The first qualitative test was obtained on 
the fifth day of incubation, that is five days before 
the first visible signs of cytological differentiation 
in the pars anterior. From the seventh day on 
enough hormone for quantitative assays could be 
found. The general results throughout develop- 
ment are tabulated below. The adult gland con- 



Age in days 


A.U. per gland 


A.U. per 100 7 
of tissue 


5 
7 


present 
0.2 






9 


0.8 




— 


12 


5.0 




9 


15 


11.0 




13 


21 (hatching) 
49 


90.0 
166.0 




60 
56 


adult 


909.0 




77 



tains about 900 A.U. which means that one gland 
has enough melanophore hormone to darken 900 
hypophysectomized lizards to stage 1 . It is of 
interest' to note the second column which shows 
the A.U. per unit weight of tissue. It may be 



July 26. 1941 ] 



THE COLLECTING NET 



seen that the concentration of the hormone per 
unit mass does not increase after hatching. All 
ai^iparent increase can be accounted for by the 
growth of the gland. It seems to suggest that 
cell for cell the pituitary reaches its peak of pro- 
duction at the time of hatching and maintains it 
there. 

But of even greater interest is the early appear- 
ance of the melanophore hormone during develop- 
ment. From analagous observations in the am- 
phibian it may very possibly be in circulation by 
the sixth day of incubation. Other hormones 
which function early in development is the thyroid 
stimulating factor of the pituitary, but it appears 
on the 11th day of incubation and the gonadotro- 
pic factors are probably not needed until later in 
development. Thus the extremely early appear- 



ance of the melanophore hormone suggests that 
its stimulation may be a very general, possibly a 
metabolic one. 

Another aspect of this prolileni concerns itself 
with the interpretation of cytological differentia- 
tion. Here we have evidence of hormone forma- 
tion and, at least in the amphibian, of hormone 
release long before we can detect or suspect such 
changes by histological methods. Such circum- 
stances must question the reliability of cytological 
interpretation of glandular function in embryonic 
tissues. 

(This work was done in collaboration with Dr. 
Painter, Dr. Kleinholz and Mr. Drager. This article 
is based upon a seminar report presented at the 
Marine Biological Laboratory on July 22.) 



THE ORGANIZATION OF THE MELANOPHORE SYSTEM IN BONY FISHES 

(Continued from page 81) 



upward therefrom, some enters the eye, passes 
obliquely upward through that structure and thus 
reaches the dorsal retina. This reflected light is 
the significant light in exciting blanching as dem- 
onstrated in several other fishes than the catfish 
by Sumner, Hogben, and especially by Butcher. 
That this is so can be shovi'n in several ways. 
When a fish is illuminated by light which is 
thrown exclusively from below so that nothing in 
the eye except the dorsal retina is effectively il- 
luminated, the animal will blanch. If the dorsal 
retina is first destroyed and the eye is then illum- 
inated from below, no blanching will occur. Fur- 
ther, if the eye is rotated on its axis through 180 
degrees so that the dorsal retina comes to lie ven- 
trally, blanching occurs only when light reaches 
this transposed dorsal organ. Thus blanching oc- 
curs only when the dorsal retina is illuminated 
and is best seen when this part of the eye is the 
exclusive recipient of light. Blanching, however, 
will take place when light reaches the ventral re- 
tina as well as the dorsal one but the change is 
not so complete as when this agent impinges ex- 
clusively on the dorsal retina. 

From the dorsal retina nerve tracts pass 
through the central nervous organs and over the 
autonomic system from which they finally emerge 
as adrenergic fibers whose terminals are in close 
proximity to the melanophores. The adrenaline 
discharged at these terminals (Chang, Hsieh and 
Lu ; and Parker) induces the concentration of 
melanophore pigment whereby the fish is made to 
blanch. Such a system of fibers may be designated 
as a retino-adrenergic arc. 

The blood of a fully pale catfish when injected 
into other catfishes either pale or dark will cause 
no change in their tints. Hence the blood of such 
fishes must be devoid of any physiological traces 
of the melanophore-expanding principle of the 
pituitary gland, intermedine : in other words the 
activity of this gland appears to be inhibited in the 



pale state of the fish. It is therefore probable that 
the dorsal retina not only excites impulses to 
blanching over the retino-adrenergic arc but also 
other impulses that check intermedine. The nerve 
tracts from the dorsal retina to the pituitary gland 
over which these impulses pass, if this is a nerv- 
ous operation, may be called collectively the reti- 
no-pittiitary inhibition arc. 

The dark phase of the catfish involves the two 
receptors, the ventral retina and the skin, and is 
best seen when the fish is on an illuminated black 
background. The action of the ventral retina is 
most conveniently studied in hypophysectomized 
fishes. The eyes of such a fish on a black back- 
ground illuminated from above receive light only 
on the ventral retina. This light is light direct 
from the source of illumination. In this case 
there is no reflected light to pass to the dorsal re- 
tina, for such light as would be reflected is ab- 
sorbed by the black background. The ventral re- 
tina thus excited gives rise to nerve impulses that 
pass through the central nervous organs and out 
over autonomic tracts to the melanophores. These 
tracts contain the cholinergic fibers which dis- 
charge acetylcholine and thus induce a dispersion 
of melanophore pigment. This dispersion is, how- 
ever, only about half that of which the melano- 
phore is capable as was first shown by Osborn. It 
may be completed by injecting intermedine into 
the fish. The tracts that reach from the ventral 
retina to the melanophores may be designated the 
retino-cholinergic arc. 

The second receptor concerned with the dark- 
ening of the catfish is the skin. This fact can best 
he demonstrated in eyeless fishes. Pale catfishes, 
catfishes of intermediate tint, and dark catfishes 
if completely enucleated and immediately put into 
perfect darkness, retain their original tint without 
change for many days. When brought out of 
darkness and into the light they quickly become 
(Continued on page 93) 



90 



THE COLLECTING NET 



[ Vol. XVI, No. 142 



The Collecting Net 

A weekly publication devoted to the scientific work 
at marine biological laboratories. 

Edited by Ware Cattell with the assistance of 
Boris I. Gorokhoff and Judy Woodring. 

Entered as second-class matter, July 11, 1935, at 
the U. S. Post Office at Woods Hole, Massachusetts, 
under the Act of March 3, 1879, and re-entered, 
July 23, 1938. 



M.B.L. CLUB NOTES 

The second '"Mixer" of the season will be held 
this evening, beginning at 8:30 P. M. Designed 
primarily to help the newly arrived students in 
invertebrate zoology to meet other biologists at 
Woods Hole, the Mixer will consist of a social 
hour, followed by dancing in which the Paul 
Jones and other dances will help to "mix" people. 
An unusual feature of the decorations will be 
aquaria with living animals generously provided 
by the Supply Department. Refreshments will be 
served. Name cards, with students' names let- 
tered on them, are being made by a committee 
consisting of Mary Chamberlain, Dr. Perry Gil- 
bert and Dr. James Goldinger. 

The Annual Meeting of the M.B.L. Club was 
held Monday evening at 7 P. M. Mr. C. L. Claff 
was reelected President of the Club ; Dr. Sears 
Crowell. who had been Secretary-Treasurer, was 
made Vice-President : and Mrs. Elsa Keil Sichel 
was elected Secretary-Treasurer. Mr. F. M. 
MacNaught was elected Trustee on behalf of the 
Club, and Dr. E. R. Clark was made Trustee on 
behalf of the Laboratory. Reports were presented 
for the House Committee by Mrs. Karl Wilbur, 
its chairman, and for the Social Committee bv 
Mrs. T. H. Bullock. The Constitution of the Club 
was amended to provide for a sinking fund for 
repairs to the Club House. 

The "Poverty Dance" held at the Club House 
last Saturday night was thoroughly enjoyed and 
well attended. Persons attending were dressed in 
old and bizarre costumes, and prizes were award- 
ed for the best outfits ; an entertainment program 
was presented which was followed by square 
dancing. The committee to judge the costumes, 
consisting of Dr. W. W. Ballard, Dr. James 
Goldinger, Miss Mary Chamberlain and Mr. Ar- 
thur Woodward, awarded prizes to the following 
persons : Mrs. Sears Crowell, for the most beau- 
tiful costume, Mrs. Karl Wilbur, for the most un- 
usual costtime, with honorable mention to L. 
Gilman, and Mr. C. L. Claff. for the "stupidest" 
costume. 

Entertainment, under the direction of J. P. 
Trinkaus, was presented with Teru Hayashi as 
master of ceremonies. The program opened with 
songs by the "Embryology Trio," composed of 



Tom Morgan, tenor, Irving Plough, baritone, and 
Jack Gross, bass. Then came the "Three Blind 
Lice," consisting of Hermann Rahn, Mrs. Jean- 
ette Renshaw, and Bob Knapp ; songs by Arlene 
Mothes ; a jitterbug dance by Dick and Jane 
Henry ; and a skit in which Frank Hartman, Bob 
Harrison and J. P. Trinkaus took part. 

The Decorations Committee consisted of Miss 
Mary Chamberlain, Mrs. C. L. Claff, Mrs. Lau- 
rence Hobson, Miss Marilyn Bosworth, and Miss 
Marion Davis. The refreshments were in charge 
of Mrs. Wilbur. 

Folk dancing is being revived at the Club this 
year under the direction of Dr. S. E. Pond. The 
first meeting was held Wednesday night. Dancing- 
is scheduled to take place on Wednesday evenings 
at 7 :00 P. M. 

Play for the annual ping-pong tournament at 
the Club will start on Monday, August 11. All 
persons interested should see Teru Hayashi for 
details. The ping pong table will be available 
until that date for training and practice. 

The ]irogram for the weekly phonograph record 
concert at the M.B.L. Club next Monday is as 
follows : Resnichek, "Overture to Donna Diana" ; 
Schumann, "Concerto in A for Piano and Orches- 
tra" ; intermission ; Mozart, "Symphony No. 40" ; 
Sibelius, "Finlandia." 



Dr. Henry B. Bigelow, curator of oceanog- 
raphy at the Museum of Comparative Zoology at 
Harvard University, was awarded the honorary 
degree of Doctor of Science at the commencement 
exercises at Yale University on June 18. He was 
formerly director of the Woods Hole Oceano- 
graphic Institution and is now one of its trustees. 

Dr. Robert Cushman Murphy, curator of 
oceanographic birds at the American Museum of 
Natural History, received the honorary degree of 
Doctor of Science from Brown University on 
June 16. Dr. Murphy lectured at the Marine 
Biological Laboratory in 1937 on "The Gates of 
the Antarctic." 



CURRENTS IN THE HOLE 

At the following hours (Daylight Saving 
Time) the current in the Hole turns to run 
from Buzzards Bay to Vineyard Sound: 
Date A. M. P. M. 

July 25 5:14 5:27 

July 26 5:57 6:12 

July 27 6:40 6:58 

"July 28 7:25 7:48 

"July 29 8:13 8:40 

"July 30 9:04 9:36 

July 31 9:58 10:36 

August 1 10:55 11:38 

In each case the current changes approxi- 
mately six hours later and runs from the 
Sound to the Bay. 



July 26, 1941 ] 



THE COLLECTING NET 



91 



ITEMS OF INTEREST 



The formal classwork for students in inverte- 
brate zoology at the Marine Biological Laboratory 
began on Friday morning when Dr. Waterman 
lectured on protozoa. They registered and had 
their desks assigned to them during the previous 
afternoon. On Thursday evening the director of 
the course. Dr. T. Hume Bissonnette, met the 
class to give them an introductory talk concerning 
matters of general interest. 

Dr. Perry W. Gilbert, instructor in zoology at 
Cornell University, has replaced Dr. Samuel A. 
Matthews as junior instructor. The laboratory 
assistant this year is Dr. J. W. Bowen, assistant 
professor of zoology at the University of North 
Carolina. 

Fifty-five students — the maximum number that 
can be accommodated — are taking the course ; 
many more made application for admission. The 
class remains in session through August 30. 

Dr. Otto Glaser, professor of biology on the 
E. S. Harkness Foundation at Amherst College, 
has been appointed acting president of Amherst 
College in the absence of the president. 

Dr. David R. Goddard, assistant professor of 
botany at the University of Rochester, and in- 
structor in the Botany course at the Marine Bio- 
logical Laboratory, has l^een promoted to an asso- 
ciate professorshi]) at the University. 

Dr. James D. Hardy, formerly research asso- 
ciate of the Russell Sage Institute and now on ac- 
tive duty in the U. S. Navy, has been appointed 
assistant professor of physiology at Cornell Uni- 
versity Medical College. 

Dr. J. S. Rankin, Jr., who has been instructor 
in biology at Amherst, has been appointed to an 
assistant professorship of biology at the Univer- 
sity of Washington. He is teaching in the inver- 
tebrate course at the Marine Biological Labora- 
tory. 

Dr. T. H. Bullock has received a Rockefeller 
Foundation Fellowship for the coming academic 
year, and will work in the department of experi- 
mental neurology at Yale University. Last year 
he was at Yale on a Sterling Fellowship in zool- 
ogy. 

Dr. George E. Coghill, member of the board 
of advisors of the Wistar Institute, died in 
Gainesville, Florida, on Wednesday at the age of 
69. He had been professor of comparative ana- 
tomy at the Wistar Institute from 1925 to 1935, 
and was managing editor of The Journal of Com- 
parative Neurology from 1927 to 1933. 



Dr. and Mrs. Charles Packard will be at 
home to members of the Laboratory tomorrow 
from four-thirty to six o'clock. 

Mr. Ray Watterson will marry Miss Evelyn 
Goddard next week in Boston. After the cere- 
monies, the couple will return to Woods Hole, 
where Mr. Watterson will continue his work. 
They will be at the Johns Hopkins University 
during the next academic year. 

Dr. and Mrs. Herbert H. Brown are spend- 
ing a few days at the Bureau of Fisheries Resi- 
dence as the guests of Dr. P. S. Galtsoff. Dr. 
Brown has just completed an appointment as di- 
rector of the sponge fishery investigations of the 
Bahama Islands. He is a member of the staff of 
the British Colonial Fishery Service, and is now 
awaiting instructions. 

Mr. H. I. Anderson, business manager of bio- 
logical Abstracts, visited Woods Hole for two 
days early this week. 

Dr. and Mrs. P. B. Armstrong have recently 
become the parents of a son. Dr. Armstrong will 
give the Friday evening lecture on August 8 and 
will work at the Laboratory for several weeks. 

The Embryology course of the Marine Biologi- 
cal Laboratory ended on Tuesday, the Physiology 
course on Wednesday and the Botany course ends 
today. 

The Atlantis, after returning from a trip under 
the direction of Dr. Henry C. Stetson, sailed on 
July 21 to Brooklyn, where she will be in dry- 
dock for several days. The Anton Dohrn is still 
out on a trip under the direction of Mr. F. Fug- 
lister. 

At the weekly staff meeting at the Woods Hole 
Oceanographic Institution on Thursday evening, 
Fred B. Phleger, Jr., of Amherst College spoke 
on "The Use of Foraminifera in Determining 
Marine Sediments." 

The Yalden Sundial, on the shore opposite the 
Marine Biological Laboratory, was damaged re- 
cently by vandals who pried off the bronze plate 
carrying the chart of instructions. The Labora- 
tory has taken steps to replace the plate. 

The J. B. Lippincott Company is holding an 
exhibit of books at the Old Lecture Hall ; the 
Clay-Adams Company closes its exhibit there to- 
day. The Bausch and Lomb Optical Company is 
continuing its exhibit at the Coast Guard Canteen. 
The Blakiston and Saunders Companies are ex- 
hibiting their publications in the Lobby of the 
Brick Building. 



92 



THE COLLECTING NET 



[ Vol. XVI, No. 142 



PHYSIOLOGY CLASS NOTES 



We have become so micro-minded lately that 
even the news this week seems to be all on the 
micro side, quantitatively as well as qualitatively. 
What with a micro Van Slyke (of Hal Gordon's 
design), microanalyses, and other things micro 
going on, we bid fair to present a micro column. 

On Monday Dr. S. E. Hill spoke on "The Ac- 
tion Current of Nitella." The subject of Dr. E. S. 
G. Barr6n"s lecture on Tuesday was "'Oxidation- 
Reduction Systems in Cellular Respiration." Dr. 
D. Wrinch talked on "Protein Structure" on 
Wednesday. 

With the breakage derby finished we have 
turned from glassware to records in our destruc- 
tive endeavors, and have succeeded in shattering 
a long victory record held by the crew in base- 
ball. The score: 7-4. A mention of stellar per- 
formances would be a roster of the entire team, 
including the faculty representatives, Drs. Kemp- 
ton and Fisher. 

Total immersion came, as it must to all kibit- 
zers, last week to one Jasper P. Trinkaus. On 



the occasion of the class photograph, his fourth of 
July left-overs spoiled a couple of fine poses. His 
subsequent entrance into Eel Pond was unfor- 
tunately not photographed for posterity. In a way 
it was a Pyrrhic victory, for our own John Gregg 
was dragged in, too. The Philip Morris repre- 
sentative, taking advantage of the occasion, passed 
cigarettes out among the crowd on the dock, 
making the incident e\-en more worthwhile. 

You have probably already glanced over our 
scientific looking little group of physiologists on 
the inside front cover and now- realize just what a 
physiology student should look like. Special men- 
tion should be made of Dr. Fisher, who appears 
to be expecting something from heaven. 

The high tide of the summer occurred on Wed- 
nesday when the course was ofi^icially over. It 
was caused by the salt tears of all the physiolo- 
gists who, in deep sorrow, packed up and checked 
out. homeward bound, having finally reached the 
end of a much enjoyed and long-to-be remembered 
five weeks. — The ex-Mr. and Mrs. J. B. V. S. 



BOTANY CLASS NOTES 



If you've got imagination and would Hke to have 

some fun, 
Just open all the stops and cocks and let it wildly 

run . . . 
We'll show you whom the Hole will miss when this 

year's course is done. 

Sam, Sam, the algology man. 
Plays hookey from lab whenever he can; 
He winks at the' women and pitches for crew. 
What kind of a guy is he? We wish we knew! 

Nancy, Nancy, with ideas romancy. 

Dodging the romeos flocking her door; 

Though she may have a cock of the head that's quite 

pert, 
We really can't picture this girl as a flirt. 

Can you see Robert Muir without his curly hair. 
Sans cigarette holder and supercilious air? 
Not resting on a bucket in the middle of the drink. 
Or collecting his algae at home in the sink? 

With a falsetto guffaw, algologist Abbott 
Wouldn't be our chuckling "Peter Rabbit", 
And if, without puffing, he kept up with Taylor, 
At the end of a field trip, he'd be even paler. 

Without "Tell me more" eyes on a Saturday nite. 
With her windblown hair always combed just right, 
You wouldn't know Babs of the red peely nose. 
Who daily for "weeds" to the drugstoi-e goes. 

Imagine gruff Monti without his sly smile. 
Not being dead serious and ribbing the while. 
And telling the girls that he will dare 'em. 
To leave the lab and join his harem. 

Picture Connie Stanton with tresses raven black, 
Deigning — at Muir's silly cracks — to even answer 

back. 
Or robbed of personality, without that sparkling 

vim, 
Not dashing from the lab each day to follow some 

fool whim. 



Picture Felix keeping quiet and not forever talking, 
See her going on field trips not complaining of the 

walking. 
Picture her without a question, silent and demure, 
Managing remarks from Monti to patiently endure. 

Imagine Ruth Franz not a quiet, sweet, girl. 

Can you see her going round in a wild, frantic 

whirl ? 
Rebelling 'gainst orders to "Go get my jacket!" 
Can you picture her ever making a racket? 

Imagine Jean Enzenbacher far, far less serious. 
Demanding some scissors with voice most imperious. 
Picture the girl never catching a cold. 
Can you see her frivolous or terribly bold? 

If Axel has shoes on, it's something quite serious, 
Can you picture him coming with shoes on the 

"Nereis" ? 
Without his hip boots and his swordfisher's cap, 
Or steering a course with the aid of a map? 

Can you picture Robert Thorne without his air-con- 
ditioned pants. 

Not starting water fights or out a-heckling, say, 
Ruth Franz? 

Or making accusations of imaginations rare, 

When the girls find parts of algae that he didn't 
think were there ? 

Picture Dr. Smith, the tall, a mere five feet, no more, 
Not beaming with that friendly grin, but getting 

really sore; 
Or picture him within the mire of lowly family 

strife, 
'Cause work in lab had made him late to luncheon 
with his wife. 

Imagine Bob without his Betty at the Mess for 

lunch. 
And picture Williams not the clown in any sort of 

bunch; 
And picture him a chemist with organic chem down 

cold — 



July 26, 1941 ] 

Without alginic acid, he's not the Bob of old. 

Imagine anything collectable — by cracky, 
Not being collected by our Babe Ruth Jackie; 
Without flowers in hair or starting a game. 
It's surely not Waldron who's so all-fired insane. 

Imagine some of us going to the bother. 
Of fearing the wrath of the tolerant Father, 
And doing our swearing completely in Spanish, 
Lest the culprits from Botany lab he should banish. 



THE COLLECTING NET 



93 



Picture Augusta without the Victoria, 
Harrietto, Antonia, and Leuchs in there, too; 
Not puffing her corncob each nite after tea, 
Wearing hair that's unkempt and expression that's 
blue. 

If you could do as we've suggested and imagine this 

strange crew. 
You'll be as glad as we are sad that this Botany 

course is thru. 

— /. W. and C. S. 



EMBRYOLOGY CLASS NOTES 



On Wednesday, July 16th, Dr. Hamburger 
considered the Molluscan larval stages omitting 
atypical Loligo studied earlier in the course for 
attention to the more typical members, Crepidula 
and Teredo. 

On Thursday the speaker's table had its heavi- 
est use of any day in the course for it supported 
the references and materials for three solidly 
packed lectures for three sure scientists. In the 
morning. Dr. Hamburger started the series with 
a review of past and recent experimental work in 
the emljryology of the Annelida and the Mollusca. 
In the afternoon Dr. Walter Landganer's lecture 
on disproportionate dwarfism in the chicken came 
as a surprise to the class. 

In the evening Dr. Harold Plough lectured on 
"Genes in Development" to an extra-large audi- 
ence. Along with many, many other facts Dr. 
Plough made it clear that the time of gene action 
varies. 

Dr. Ballard took over the course on Friday for 
the presentation of the final materials of the 
course. This day it was wet Coelenterates and 



they balanced very well with much dry humor. 

Saturday started out to be a disappointing day 
for the embryologists for their scheduled towing 
was cancelled due to high winds from an unfavor- 
able direction. Yet this was soon forgotten when 
examination of tows obtained earlier in the day 
was begun. Every marine biologist, manual, and 
textbook in sight was practically worn out in two 
hours, so anxious was the class to learn about 
oddities they discovered. 

Dr. Caswell Grave gave the last formal lecture 
of the course on Tuesday morning. His subject 
was metamorphorsis in the Ascidians. Throug- 
out the lecture Dr. Grave said everything to nul- 
lify his preliminary slogan, "Work with Ascidians 
and be lonesome." Conditions favoring, I am sure 
that Dr. Grave would have had thirty-seven co- 
workers at the end of his lecture. By afternoon 
the students had l^egun to depart. Happy for 
having lived five such edifying weeks, sad for hav- 
ing to part from such good friends, the students 
of the 1941 Embryology Class went their scat- 
tered wavs. — E. R. 



THE ORGANIZATION OF THE MELANOPHORE SYSTEM IN BONY FISHES 

(Continued from page 89) 



coal-black. The lilood from these fishes when in- 
jected into pale fishes will darken the recipients, 
for it contains intermedine. Apparently the pho- 
toreceptors of the catfish skin when excited by 
light give out impulses that pass through the cen- 
tral nervous organs to the pituitary gland which 
is thereby excited to discharge intermedine. This 
in turn is carried by the blood to the melano- 
phores which respond by complete pigment dis- 
persion. If the brain of a catfish is completely 
transected immediately in front of the cerebellum, 
the eyes and the pituitary gland are left intact on 
the anterior part of the central nervous fragment 
and the whole skin inner-\'ation remains unaltered 
on the posterior part. Catfishes that have under- 
gone this operation show no obvious color 
changes, which indicates two important conclu- 
sions ; first, that the eyes of this fish are not con- 
cerned with exciting the discharge of intermedine 
from the pituitary gland, and, second, that the 
darkening of the skin in the catfish is not a spinal- 
cord reflex. The tracts by which the skin photo- 
receptors in the catfish are connected with the pi- 
tuitary gland and the blood courses by which the 
intermedine from this gland is carried to the me- 



lanophores may be called collectively the dermo- 
pituitary arc. 

Of the bony fishes whose color changes have 
been studied within the last few years the follow- 
ing appear to conform in general to the type of 
chromatic organization described for the catfish 
in so far as they possess both adrenergic and 
cholinergic fibers and intermedine : angelfish 
(Tomita), eel (Waring), snakefish (Chang, 
Hsieh and Lu), Japanese catfish (Matsushita), 
and stickleback (Hogben and Landgrebe). How- 
ever, in none of these instances is it known that 
the skin acts as a receptor in the way that it does 
in the catfish. Certainly in the killifish and prob- 
ably in flatfishes (Osborn) adrenergic and choli- 
nergic fibers are present, but intermedine appears 
to play a very subordinate part in these forms. 
Further study will probably show that the chro- 
matic systems of different bony fishes are speci- 
fically individual rather than that they conform 
to a single type of organization. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on 
July 22.) 



THE COLLECTING NET 



Vol. XVI, No. 142 



SUPPLEMENTARY DIRECTORY FOR 1941 



ADDITIONAL INVESTIGATORS 

Aquila, (Sister) M. grad. biol. Villanova. Rock 3. 

Beck, L. V. instr. phys. Hahnemann, lib. 

Boche, R. D. instr. zool. Pennsylvania. Br 221. D 

112-A. 
Bodian, D. asst. prof. anat. Western Reserve Med. 

lib. 
Bronfenbrenner, J. prof. bact. & immun. Washing- 
ton Med. (St. Louis). Br 305. 
Brown, D. E. S. prof. phys. New York. Br 310. 
Calkins, G. N. prof, proto. Columbia. Br 331. 
Castelmano, Gina zool. Minnesota, lib. 
Chambers, E. Nevi^ York Med. Br 343. 
Chambers, R. prof. biol. New York. Br 328. 
Clark, Eleanor L. assoc. anat. Pennsylvania Med. Br 

117. 
Duncan, G. W. fel. surg. Hopkins. Br 328. 
Erianger, Margaret instr. West Virginia. Br 312. 
Finkel, A. J. grad. asst. zool. Chicago. Br 322. 
Garner, H. res. asst. zool. Chicago. Br 332. Ka 22. 
Gelback, Elizabeth L. asst. proto. Yale. Br 323. 
Genevieve, (Sister) Mary grad. biol. Villanova. Rock 

3. 
Gilbert, P. W. instr. zool. Cornell. OM. 
Goldinger, J. M. res. asst. med. Chicago. Br 125. K 

14. 
Grand, C. G. res. asst. biol. New York. Br 311. 
Gurewich, V. Cornell Med. lib. 
Hamilton, H. L. res. asst. emb. Hopkins. Br 324. 
Harnly, M. H. assoc. prof. biol. New York. Br 344. 
Horn, Annabelle grad. asst. zool. Pittsburgh. Rock 7. 
Hunter, G. W., Ill asst. prof. biol. Wesleyan. 
Keefe, E. L. res. asst. zool. Washington (St. Louis). 

Br 217-j. 
Klotz, J. W. grad. zool. Pittsburgh. Rock 7. 
Kopac, M. J. asst. prof. biol. New York. Br 311. A 

106. 
Kreezer, G. L. asst. prof, psych. Cornell, lib. D 312. 
Lancefield, D. E. assoc. prof. biol. Queens (New 

York). Br 126. 
Lorenz, P. B. Swarthmore. OM Phys. Ho 3. 
Lucas, A. M. assoc. prof. zool. Iowa State. OM 29. 
Marvel, R. (Bartlett, N. H.). OM 21. D 211. 
Mead, F. W. Ohio State. Br 111. 
Metz, C. W. prof. zool. Pennsylvania. Br 304. 
Michaelis, L. mem. Rockefeller Inst. (New York). 

Br 207. 
Morgan, Isabel M. asst. path. & bact. Rockefeller 

Inst. (New York). Br 320. 
Mullins, L. J. asst. phys. Rochester. Br 322. 
Netsky, M. Pennsylvania Med. Br 205. 
Northrop, J. H. mem. Rockefeller Inst. Med. Res. 

Br 209. 
Pick, J. instr. anat. New York Med. Br 343. 
Rabinowitch, E. res. assoc. M.I.T. lib. 
Rahn, H. instr. zool. Wyoming, lib. 
Reiner, J. M. biophysics (New York, N. Y.). lib. 
Renshaw, B. asst. prof. zool. Oberlin. Br 218. 
Sandow, A. asst. prof. biol. New York. Br 344. 
Spratt, N. T., Jr. res. asst. emb. Hopkins. Br 324. 
Stebbins, R. B. grad. asst. biol. New York. Br 328. 
Stewart, Dorothy R. assoc. prof. biol. Skidmore. Br 

205. D 316. 
Stiegelman, S. asst. zool. Columbia. Br 313. 
Sturtevant, A. H. prof. biol. California Tech. Br 126. 
Trager, W. assoc. Rockefeller Inst. (Princeton). Br 

208. 
Turner, Abby H. prof. phys. Mt. Holyoke. lib. 



Warren, A. A. asst. path. Harvard Med. L 27. 
Weaver, Margaret A. grad. zool. Texas. Br 312. 
Wilde, C. E., Jr. instr. zool. Dartmouth. OM 40. 
Zimmerman, G. L. Swarthmore. OM Phys. Ho 3. 
Zingher, J. M. C.C.N.Y. Br 315. 

STUDENTS IN INVERTEBRATE ZOOLOGY 

Anderson, Dorcas J. grad. asst. biol. Purdue. K 2. 

Andrews, T. J. asst. zool. Massachusetts State. 

Aram, H. H. grad. zool. State U. Iowa. 

Batchelor, W. H. Harvard. 

Beardsley, Margaret Smith. WC. 

Berg, W. grad. asst. zool. State U. Iowa. 

Brainerd, J. W. grad. biol. Harvard. 

Brown, Henrietta B. Tufts. WB. 

Burke, R. K. asst. biol. Springfield. Dr 8. 

Byerrum, R. Wabash. Ka 22. 

Carpenter, Elizabeth grad. asst. zool. Mt. Holyoke. 

WE. 
Cole, L. C. grad. Chicago. 
Corder, H. R. Williams. Dr 10. 
Cornish, Helen R. teacher biol. Virginia Intermont 

(Bristol, Va.). 
Culberson. A. W. Williams. 
Dodd, S. G. Weslevan. Ki 5. 
Dole, Dorothy K. Bates. H 7. 
Garman, Elizabeth M. New Jersey Col. Women. 

WG. 
Gillette, R. J. res. asst. zool. Washington (St. Louis). 

Dr 5. 
Gilligan, Catherine teacher biol. Hyde Park H. S. 

(Mass.). 
Goffin, Mary F. Seton Hill. 
Hahn, Rhea J. Radcliflfe. H 1. 
Harris, N. D. teacher Berkshire. Dr 2. 
Hauschka, T. S. grad. zool. Pennsylvania. 
Heaps, Marian E. grad. biol. Lebanon Valley (Pa.). 
Hinde, H. P. grad. asst. zool. Yale. Dr 2. 
Humm, D. G. grad. zool. Yale. D 311. 
Kielich. E. R. asst. biol. Canisius (Buffalo, N. Y.). 

Dr 2. 
King, Ellen E. asst. biol. Sarah Lawrence. H 7. 
Kohler, C. E. Rutgers. Dr 1. 
Lumb, Ethel S. grad. asst. zool. Missouri. WD. 
Mahr, M. M. grad. asst. biol. New York. 
Mead, A. R. grad. asst. zool. California. Dr Attic. 
Miller, Helena A. grad. biol. Radcliffe. WC. 
Miner, H. D., Jr. asst. zool. Wabash. Ka 24. 
Osmun, J. V. grad. biol. Amherst. Ka 22. 
Paull, J. grad. biol. Harvai'd. Ka 2. 
Perkins. D. D. Rochester. Dr 1. 
Pond, S. M. asst. biol. Wesleyan. Ki 5. 
Powers, W. T. instr. zool. De Paul. 
Randall, W. C. grad. asst. biol. Purdue. K 7. 
Roberts, Beryl J. teacher Trade School (Boston). 
Roberts, H. S., Jr. grad. asst. zool. Duke. 
Roberts, W. F. grad. asst. zool. Northwestern. 
Robinson, Margaret H. Wellesley. H 7. 
Ross, Lucille Barnard. 
Schlichter, Helena L. Wilson. H 1. 
Senyard, Juanita asst. biol. Oberlin. H 7. 
Talmage, R. V. N. instr. zool. Richmond. K 15. 
Tuttle, Ruth F. Wheaton. WI. 
Weber, Ann M. Montclair Teachers. WB. 
Wieder, H. Hamilton (N. Y.). 
Wilber, C. G. grad. asst. zool. Hopkins. K 15. 
Williams, R. W. Harvard. 
Wilson, Mae E. grad. zool. Southern California. K 3. 



July 26, 1941 ] 



THE COLLECTING NET 



95 




First to build the Barcroft- 
Warburg Apparatus in the 
United States, the J. H. Emer- 
son Co. has now developed 
this greatly improved model. 

Features include a full range 
of speed adjustment ; conveni- 
ent adjustment to obtain any 
amplitude of shaking, with 
provision to bring manometers 
to vertical position for read- 
ing ; motor and drive mounted 
under tank. 



EXHIBIT AT WOODS HOLE 

Old Lecture Hall, August 1 to 11 



In 1928, the J. H. Emerson Company was 
established manufacturing experimental ap- 
paratus, and in 1931 developed the modern, 
diaphragm-type Respirator ("Iron Lung"). 
Since that time, this company has grown 
yearly, designing and manufacturing hospi- 
tal equipment, and, in the last year, pro- 
duced more "Iron Lungs" than any other 
manufacturer. 

Due to our desire to continue our con- 
tacts with those who are doing research, we 
recently set up a new shop to be devoted 
entirely to the designing and making of 
experimental apparatus. 



In this manner, we will be able to con- 
tinue the manufacture of Warburg Ap- 
paratus, the Emerson Micromanipulator, 
high-speed centrifuges, etc., as well as 
special apparatus to the customer's re- 
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Mr. John Linden, who is in charge of 
this development shop, will be at our 
Woods Hole exhibit, and will be glad to 
discuss special problems and demonstrate 
our Warburg, Manipulator, and centrifuge 
equipment. 



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"Iron Lungs" — Fever Cabinets 
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CAMBRIDGE, MASSACHUSETTS 



96 



THE COLLECTING NET 



[ Vol. XVI, No. 142 



Cambridge Instruments 

Cambridge archives could yield interesting 
stories of cooperation with many notable 
scientists. From its inception this company 
has specialized in making precision instru- 
ments for exacting professions. As a result 
the name "Cambridge" is a respected one in 
Science, Medicine and Industry, and there 
are few fields in which Cambridge instru- 
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Camljridge products include Galvanometers, 
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Write for literature of instruments of use 
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This galvanometer is an inexpensive instru- 
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and is accordingly well adapted for general 
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and a reflecting mirror suitable for "null" 
point indications as well as for use with a lamp 
and scale. It is only one of many Cambridge 
Galvanometers. 



HUMAN SKULLS 

For several >-ears it has been increasingly difficult to obtain good 
skeletal material. Just recently, however, (through a fortunate con- 
nection with a collector in India) we purchased a lot of nearly ISO 
skulls, most of which were of unusually fine quality. These are now 
offered at prices of from $14.00 to $35.00 depending upon the com- 
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It will pay an\- school to take advantage of this offer in providing 
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General Biological Supply House 

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761-763 East Sixty-Ninth Place, Chicago 

The Sign of the Turto.v Pledges Absolute Satisfaction 




July 26, 1941 ] 



THE COLLECTING NET 



97 



Kern Scientific Instruments Since 1819 



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Booksize slide boxes are about 2'/2" shorter than the usual 
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98 



THE COLLECTING NET 



[ Vol. XVI, No. 142 



Adams UTILITYFORCEPS Stainless Steel 
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July 26, 1941 ] 



THE COLLECTING NET 



99 



A New Spencer 
COLORIMETER 

O Simplified Technique 

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9 Spencer annuuiices a LJiillI Result 
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100 



THE COLLECTING NET 



[ Vol. XVI, No. 142 






LIGAMENTS 




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Vol. XVI, No. 6 



SATURDAY, AUGUST 2, 1941 



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EFFECT OF SEA WATER ON THE RADIO- STRUCTURE OF THE RED CELL IN THE 



SENSITIVITY OF ARBACIA SPERM 

Drs. T. C. Evans and J. C. Slaughter 

Dcpartiiiciifs of Radiology ami Zoology, 

State Uuifcrsity of loiva 

In the course of an investigation of the effects 
of roentgen radiation on the oxygen consumption 
of Arbacia sperm it was noted that dilute sus- 
pensions of sperm were in- 

jured more by the radiation 
than were sperm irradiated in 
concentrated suspensions. This 
observation was checked by ir- 
radiating sperm in different 
concentrations of sea water 
and later determining the fer- 
tility of each lot. This was 
done by adding the same 
amounts of control and irra- 
diated sperm to similar lots of 
eggs. The percentage fertili- 
zation was taken as the num- 
ber of eggs which raised ferti- 
lization membranes per hun- 
dred counted. In a series of 
five experiments consistent re- 
sults were obtained which in- 
dicated that as more sea water 
was added to the sperm their 
radiosensitivity increased. The 
results of one of these experiments are shown in 
Table I. 



% % (Ealenliar 



TUESDAY. August .5, 8:00 P. M. 

Seminar: Dr. Victor Schechter: 
"Aging Phenomena, and Factors 
Influencing the Longevity of 
Mactra Eggs." 

Dr. Frederick S. Philips: "Com- 
parison of Regional Respiratory 
Rates of the Chick Embryo dur- 
ing Early Stages of Develop- 
ment." 

Dr. Matilda M. Brooks: "Further 
Interpretations of the Eff'ects of 
CO and CN on Oxidations in Liv- 
ing Cells." 

FRIDAY, August 8, 8:00 P. M. 
Dr. P. B. Armstrong: "Function is 
the Developing Gastro-Intestinal 
Tract of Amblystoma punctatum 
in Relation to Embryonic Deter- 
mination and Difl^erentiation." 



LIGHT OF SHAPE TRANSFORMATIONS 

Dr. Eric Ponder 
The Nassau Hospital. 
Mineola, New York 

Since the circular, biconcave form of the mam- 
malian red cell was first correctly described by 
Hodgkin and J. J. Lister in 1827 (over ISO years 

after the discovery of the cell 

by Swammerdam and later by 
Leeuwenhoek) there has been 
an almost continuous specula- 
tion as to the reason why this 
special shape should be as- 
sumed by bodies floating free- 
ly in a liquid. Generally 
speaking, the theories which 
have been advanced have been 
of two kinds : those which at- 
tribute the biconcave shape to 
forces or structures in the in- 
terior, and those which attri- 
bute it to forces or structures 
at the surface. 

Into the first category falls 
the "gelatin lozenge" theory of 
Rollett (1862) "who conceives 
that a stroma or matrix enters 
into the structure of the color- 
less elastic extensible substance 
of the red corpuscle, and that to this the form and 
the peculiar physical properties of the corpuscles 



It will be noted also {Continued on page 113) is due. It is supposed that the coloring matter 



Structure of the Red Cell in the Light of 
Shape Transformations, Dr. Eric Ponder.... 101 

Effect of Sea Water on the Radiosensitivity 
of Arbacia Sperm, Dr. T. C. Evans and 
J. C. Slaughter 101 

Some Aspects of Pigment Deposition in 
Feather Germs of Chick Embryos, Dr. Ray 
L. Watterson 107 



TABLE OF CONTENTS 

The Influence of Hormones on the Differen- 
tiation of Melanophores in Birds, Dr. H. L. 

Hamilton 109 

Conference at the University of Chicago 110 

Items of Interest Ill 

Studies on the Life History of Siphodera 
vinaledwardsii, a Trematode Parasite of the 
Toadfish, Drs. R. M. Cable and A. V. 

Hunninen 112 

Invertebrate Class Notes 113 




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bJ ^ 
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August 2, 1941 ] 



THE COLLECTING NET 



103 



can be separated from the stroma without causing 
the latter to lose its essential characters" (quota- 
tion from Norris, 1882; Thudicum, writing at the 
same time, goes further, and defines the stroma 
as a chemical skeleton, with which the hemoglobin 
is combined, an idea which has been revived by 
Lepeschkin, Adams, and others). Gough and 
Teitel-Bernard have suggested that molecules in 
the cell interior, and particularly hemoglobin mol- 
ecules, repel one another more in one axis than 
in another, and so give rise to the discoidal shape 
in a cell with a fluid interior, but this hypothesis 
cannot be held in view of the fact that hemoglo- 
bin-free ghosts are discoidal. 

The second point of view is by far the older, 
and although it is associated with the names of 
Schwann and of Hewson, it was originally ex- 
pressed by Bidloo in 1685 and by Wells in 1797. 
It was emphatically defended, particularly against 
the view of Rollett, by Schafer (1891), whose 
description of the cells as "vesicular bodies pos- 
sessing an external envelope enclosing a fluid in- 
terior" has come to be known as the "balloon 
theory". In 1882, Norris was impressed, as sev- 
eral others have been (Rice, 1914, Gough, 1924), 
by the similarity of shape between red cells and 
the myelin forms of lecithin. The latter are often 
circular discs about 5^ to lO/i in diameter, and 
are dumb-bell or ring shaped in cross section. 
They are apparently formed by physical forces at 
the interfaces between the droplets and the fluid 
surrounding them, and Norris suggested that the 
biconcave shape of the mammalian red cell is 
brought about in a similar way. "The remarkable 
properties displayed by myelin at once relieve us 
from the necessity of considering that one liquid 
or solution submerged in another must inevitably 
take on globular or spherical state. The fact is, 
the substance appears to represent the extending 
or spreading-out tendency, as opposed to the 
gathering-up or sphere-forming property. The 
biconcave and the annular forms seem to be re- 
lated to a kind of balancing of these two proper- 
ties. ... I consider, too, that the form which these 
bodies assume is as dependent on the constitution 
of the liquid in which they are submerged as on 
their own. ... It would therefore seem that the 
biconcave form is to be regarded as an arrested 
annular form. This annulating property belongs 
to the corpuscle as a substance (italics mine) for 
it occurs in fused masses of corpuscles, in single 
corpuscles, and in fractional parts of them" (Nor- 
ris). Or, as Gough puts it, there are two sets of 
forces operating, the first of which tends to pro- 



duce contraction of the surface and the spherical 
form, while the second tends to bring about ex- 
pansion and a very flattened form ; balanced 
again.st each other, the two sets of forces maintain 
the discoidal form. It is not diff'icult to see how 
mutual repulsion between molecules in the surface 
layers might arise, for hydrocarbons with polar 
groups directed towards the water (as in lecithin) 
might exhibit repulsion of each other. 

Some idea of the forces involved may be ob- 
tained by drawing a cross section of the red cell 
to scale, finding the two principal radii of curva- 
ture pi and p-2 at each point, and computing the 
]3ressure P which would have to be applied to 
keep a homogeneous cell membrane, with tension 
T, in hydrostatic equilibrium : 

P = T(l/p^+l/p,). 

It appears that we have to have a pressure di- 
rected outwards over the equatorial regions of the 
cell, and a smaller pressure directed inwards over 
the biconcavities, if we are to arrive at the shape 
in this way. The idea that such pressures really 
e.xist is, of course, untenable, but the "outward 
pressure over the equatorial regions" is the same 
thing as Gough's "expansive force". What really 
hap])ens is probably a variation in the tensions 
from point to point along the membrane, and this 
is the .same as saying that the membrane is not 
molecularly homogeneous, or has a "liquid crys- 
tal" structure. 

In 1926 (Ponder, 1933) I tried to put this at- 
tractive idea into mathematical form, with a sug- 
gestive, if disappointing, result. I evaluated the 
radii of curvature for the red cell of man, and, on 
the assumption that the tensions in the membrane 
are equal at all points, obtained a series of values 
for the pressures which would maintain the shape : 
if, as is probably the case, the pressures are equal 
and the tensions vary, the true relations are de- 
rivable from the same sets of values, and any 
theory proposed for the shape of the red cell must 
be one which gives these relations, at least sub- 
stantially. I then tried to find the shape of a 
body of given volume, given area (not necessarily 
the smallest area for the volume, for then the body 
would be spherical) and with the smoothest sur- 
face, and arrived at a form suggestive of that of 
the red cell ; the derivation, however, was faulty 
in that it dealt with a mathematical surface rather 
than with a surface with real elastic properties, 
but the result was interesting in showing how a 
problem of this kind might be approached, and 



The Collecting Net was entered as second-class matter July 11, 1935, at the Post Office at Woods Hole, Mass., 
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104 



THE COLLECTING NET 



[ Vol. XVL No. 143 



what a formulation of Norris' theory might in- 
volve. 

The first prolilem, then, is to decide whether 
the form of the red cell is due to forces or struc- 
tures in the interior, or to forces or structures at 
the surface. Microscopical observation, either with 
direct illumination, with the dark field. wiUi ultra- 
violet light, or with the electron microscope, tells 
us little one way or the other about the existence 
of a stroma, and evidence from microdissection 
studies is equally inconclusive ; sometimes the red 
cell disappears altogether when punctured, but 
sometimes there is a mass left behind which can 
be teased into shreds. This may lie a product of 
gelation, for the red cell has been shown to con- 
tain proteins other than hemoglol^in, e.g.. the stro- 
matin of Joqies (1932) and of Boehm (1935). 
which is capable of forming a remarkably rigid 
gel in low concentrations. Turning to the sur- 
face, we find a distinction between the cell "wall" 
(ir "envelope" and the cell "membrane", the for- 
mer lieing at least thick enough to be visible 
{ 0.05/i ) , while the thickness of the latter has been 
l)ut at anywhere from 30 A. to 300 A. or 
more. We have no indication as to how forces 
responsible for the shape of the cell might arise 
at such a surface, and both Rollett's theory and 
Xorris' theory demand some kind of special struc- 
ture (a "stroma", or a special molecular configur- 
ation at the surface), and we do not have much 
direct evidence that such a structure exists. We 
may accordingly turn to the shape transforma- 
tions of the red cell, and see what they tell us. 

1. Pisc-s/^licrc tnnisjoiiiiatioiis. 

.\s hitherto described, these are five in number, 
and the study of each brings out some special 
point which has a bearing on the subject of red 
cell shape. 

7. Tlic s/^licrical form bcHcccii tico surfaces. 
Mammalian red cells, suspended in 1 p.c. NaCl 
or any of the ordinary physiological salines, and 
examined in a hanging or uncovered drop, are 
discoidal although often crenated ; at all events. 
they are not spherical. If the same cells are cov- 
ered with a coverglass so that the amount of fluid 
between the two glass surfaces is very small, the 
cells become smooth spheres of the same volume 
as were the original discs. If the two surfaces 
are in close contact, the change is very rapid, but 
by constructing a wedge-shaped chamber it can 
be seen that the transformation involves inter- 
mediate forms ; the discs first crenate, the crena- 
tions becoming progressively finer until a prickly 
form, the "thorn-apple" form, is assumed ; the 
crenations then become finer still and are ulti- 
mately smoothed out, so that a glistening sphere 
results. If a small amount of serum or plasma is 



run under the coverglass, the spheres become cre- 
nated, the crenations become larger, and the dis- 
coidal form re-appears ; these re-converted discs 
may not be as perfect in shape as the original 
cells, and they are unusually sticky. Millar (1925) 
describes them as being mottled when seen with 
the dark field. 

Since the original description of the spherical 
form by Hamburger in 1895, there has been much 
s])eculation as to the cause of this transformation, 
but in 1940 Furchgott showed that it is due to 
(a) an increase in pH produced by diffusion of 
alkali from the glass surfaces, and (b) the re- 
moval from the cells of an "anti-sphering sub- 
stance", which he and I later showed to be the 
carbohydrate-poor fraction of serum albumin, or 
crystalbumin (Furchgott and Ponder, 1940). This 
substance is adsorbed from a suspension of red 
cells by glass surfaces, such as the surfaces of the 
slide and coverslip or the surface of glass beads, 
and cells freed of it will adsorb it again, quantities 
of the order of 800 mg. per 100 cc. of cells being 
involved. If all this were taken up at the red cell 
surface, it would form a layer only a few mole- 
cules thick, but such a layer would have a volume 
about one-third that of the estimated volume of 
the red cell membrane as a whole. 

2. The spherical form- produced by lecithin. A 
similar disc-sphere transformation, with thorn- 
apple forms as intermediates, occurs when lecithin 
is added to red cells suspended in serum, plasma, 
or saline. The lecithin is most conveniently used 
in the form of a sol. made by adding 0.5 cc. of a 
10 p.c. solution of lecithin in alcohol to 100 cc. of 
boiling saline. This sol keeps for weeks in the 
refrigerator, although some of the lipoid may sep- 
arate out, but the lecithin in it can be estimated 
from time to time. Again the spheres are formed 
without an}' change in volume ; they ultimately j 
hemolyse. but not for hours when minimal 
amounts of lecithin are used. I have recently 
done a considerable amount of work on the quan- 
titative aspects of this transformation, and the re- 
sults may be summarized as follows. ( 1 ) The 
quantity of lecithin required to bring about the 
disc-sphere transformation is remarkably constant 
for the cells of one species, and for washed human 
cells the amount needed is about 4 molecules per 
A-. of cell surface. This quantity is large, for the 
smallest cross-section of the lecithin molecule is 
]3robably itself about 14 A-. If the cells are sus- 
])ended in plasma, about seven-eighths of the 
lecithin is adsorbed or otherwise combined with 
the plasma proteins, and only the remaining 
eighth is available for the cells. (2) The amount 
of lecithin needed to complete the disc-sphere 
transformation is about double that required to 
initiate it ; quantities intermediate give forms with 
crenations of increasing fineness. (3) About half 



August 2, 1941 ] 



THE COLLECTING NET 



105 



the amount is needed at 37°C. as at 4°C., so the 
temperature coefficient is positive, but not large. 
(3) If tlie cells are fixed with fixatives such as 
forniol, the amount of lecithin required to produce 
spheres is increased roughly as the square of the 
formol concentration. As unfixed cells require a 
certain amount of lecithin to initiate the change 
from disc to sphere, the effect of formol is simply 
to increase this yield-point, a fact which has some 
bearing on the natural rigidity of the cell and on 
the increase of this rigidity on gelation. 

The disc-sphere transformation can be reversed 
by washing the cells in untreated plasma, serum, 
or saline, and the transformation and its reversal 
can be repeated several times under favorable cir- 
cumstances, although each repetition involves 
more and more lysis. The variations in the 
amount of lecithin required to transform the red 
cells of different animals has not been fully 
worked out, but such differences exist. 

3. The spherical joriii produced by rose beiigaL 
Red cells suspended in plasma, serum, or saline 
undergo a typical disc-sphere transformation with- 
out change in volume if small amounts of eosin 
(M/10-''). erythrosine (M/10^), or rose bengal 
(M/10^) are added in the dark, the effect on 
washed cells being much greater than on cells in 
plasma. In the light, smaller quantities of dye 
are needed, and the quantity of rose bengal which 
brings about the transformation in the rabbit red 
cell is such as would cover only about 1/20 of 
the cell surface, even if it were all concentrated 
there. The change in shape, in this case at least, 
is presumably Ijrought about by an effect on a sur- 
face component. The spheres can be re-converted 
into discs by the addition of plasma, and the active 
component seems to be plasma protein ; the re- 
converted discs show the same stickiness, slight 
crenation. and mottling as is seen in the case of 
discs re-converted from spheres between slide and 
coverglass. 

4. Spherical forms produced by other lysius. 
It is now recognized that all forms of hemolysis 
are preceded by a disc-sphere transformation, 
sometimes occurring immediately before lysis, as 
in the case of saponin, and sometimes a long time 
before, as in the case of lecithin. What happens 
seems to be that at a certain stage of the action of 
the lysin on the cell membrane a "shape compo- 
nent" gives way, and the cell, often quite sudden- 
ly, becomes a sphere ; after further action a "per- 
meability component" breaks down, and the cell 
hemolyses. Although this loss of shape and this 
loss of semi-permeability are two stages of one 
lytic ]irocess, the "shape component" can be de- 
stroyed without any important change in the per- 
meability properties. Further, there is no appre- 
ciable change in the resistance, capacity per unit 
area, or frequency dependence of the capacity. 



when the disc-sphere transformation occurs (Cur- 
tis, 1935), nor of the electrical mobility (Furch- 
gott and Ponder, 1940), which ap])ears to be un- 
changed, indeed, even after the cell is hemolysed 
in several different ways (Abranison, Furchgott, 
and Ponder, 1938). These observations lead not 
only to the conclusion that the "shape component" 
is distinct from the "permeability component", but 
that the phenomenon of lysis does not necessarily 
involve the surface of the cell membrane as a 
whole, and many diverse observations support this 
view (see Ponder, 1941, a, for a summary of the 
evidence). 

5. The spherical forui in hypotonic solutions. 
When the red cell is placed in a hypotonic solu- 
tion, it swells, sometimes as much as would be ex- 
pected if it were a simple osmometer and some- 
times less (Ponder, 1940), and in doing so be- 
comes cup-shaped or bell-shaped. If the solution 
is sufficiently hypotonic it more or less suddenly 
becomes spherical, and then hemolyses, and it has 
been shown (Ponder, 1937, Castle and Daland, 
1937) that the surface area of this sphere is sub- 
stantially the same as that of the original disc, al- 
though the sphere contains the greater volume 
(135 - 150 volumes as compared with 100 for the 
disc ) . Examination of the cells, now become 
ghosts, will reveal the fact that they are once more 
discs, of substantially the same volume as they 
were initially. The re-assumption of the discoidal 
form is ajajjarently spontaneous, the forces on the 
membrane having been relieved by the lysis of the 
cell. If sufficient salt is now added to make the 
system isotonic, the discoidal ghosts shrink to 
aljout 60 ]3er cent of their volume ("reversal of 
hemolysis" ) , showing that they retain some semi- 
permeability, but after a little while they swell 
to take up the initial volume once more (Ponder, 
1941, m.). 

The remarkable fact is that such ghosts cannot 
be turned into spheres by placing them between 
slide and coverglass, by adding lecithin, or by 
treating them with lysins such as rose bengal and 
saponin. They seem to have undergone a "per- 
manent set" in the shape of discs. 

2. The ultrastructure of the cell uieuihranc. 

The study of the disc-s])here transformations 
lead us to the conclusion that the changes are 
jirobably due to the action of substances which act 
at surfaces, and that the component which main- 
tains the shape is probabl)^ a surface component. 
This brings us back to Norris' theory, which de- 
mands a kind of "liquid crystal" structure of the 
cell surface layers. Chemical analysis shows the 
red cell membrane to be a protein-lipoid complex, 
and since both X-ray analysis and the methods of 
jxjlarization optics have shown protein-lipoid com- 



THE COLLECTING NET 



106 

plexes in other memljranes (e.g., the axon sheath, 
.Schinitt, 1936) to have a micellar organization, it 
is not surprising to find that the optical properties 
of the membranes of the ghost also show the pres- 
ence of an ultrastructure (Schmitt, Bear, and 
Ponder, 1936, 1938). This structure may be in- 
terpreted as consisting of layers of protein orient- 
ed tangentially, and layers of lipoid molecules 
(jriented radially, the phosphoric acid group of 
the cephalin molecules being towards the water 
side of the interface, and dominating the situation 
so far as the electrophoretic properties are con- 
cerned (Furchgott and Ponder, 1941). Judging 
from the quantities of jirotein and of lipoid found 
in the membrane by chemical means, there may 
he several such layers alternating with each other 
( as in the axon sheath ) , although not in the form 
of continuous films, for Parpart and Dziemian's 
(1940) figures show that the amount of extract- 
able li]ioid, all of which is contained in the cell 
membrane as we know it, is not sufficient to make 
more than a bimolecular layer 30 A. thick. The 
])rotein moiety would ])rovide layers with a total 
thickness of about 90 A., and adding the two con- 
tributions together, the thickness of the membrane 
would work out at about 120 A, 

There has been in the past, and still is, con- 
siderable doubt as to this total thickness, and this 
is partly because the analytical figures given by 
difterent investigators have not agreed very well 
with each other, partly because of an insistence on 
the necessity of continuous molecular films instead 
of a binding of phos])holipoid to protein, which 
would lead to an orientation of cephalin and le- 
cithin at particular loci around the protein mole- 
cules in the membrane ( Parpart and Dzieniian, 
1940), and partly because the contribution of 
water has not always been taken into account. Es- 
timates of the extent of this contribution range 
from 10 p.c. of the thickness to 100 p.c. of the 
thickness; Waugh and Schmitt (1940) give about 
25 p.c. Estimates of the total thickness of the 
membrane of the ghost range from 30 A. (Gorter 
and Grendel, 1926, Fricke, 1926), to 120 A. (Par- 
part and Dziemian, 1940), 120 A. exclusive of 
the contribution of water (Fricke, Parker, and 
Ponder. 1939), 135 A. at pH 7 and 220 A. al 
jdH 6 (Waugh and Schmitt, 1940), and even 
higher values. Recently Zwickau (1941) has ob- 
tained photographs of fixed and dried membranes 
of ghosts by means of the electron microscope, 
and sets the thickness as from 200 to 300 A. If 
the contribution of water is allowed for, his values 
would be among the highest yet suggested. The 
thickness of the membrane in the intact cell may, 
of course, be greater than it is in the ghost, for 
substances, perhaps not essential to the resistance, 
capacity, and semi-permeability of the cell surface 
may be washed out of it in the process of hemo- 
lysis. The anti-sphering substance, which makes 



[ Vol. XVI, No. 143 



u]) about one-third of the protein content of the 
membrane of the disc, is one such substance. I 
therefore feel that it is ])ossible that the cell mem- 
brane may approach 500 A. in thickness under 
certain circumstances, and so reach such dimen- 
sions as to be visible, although not resolvable, by 
the microscope. 

3. The interior. 

The conclusion that the red cell possesses a 
membrane with an ultrastructure and that the 
shape is governed by molecular arrangements and 
forces in the surface layers does not stand in the 
way of its possessing an interior structure as well, 
although the views of Rollett and of those who 
insisted on the complex nature of the surface have 
often been thought of as mutually exclusive. There 
are at least four good reasons for thinking that 
the interior is not occu]oied simply by a homogen- 
eous solution of hemoglobin and salts. 

1. There is a correlation coefficient of only 
about 0.5 between the density of the red cell and 
its hemoglobin content. In the absence of hemo- 
globin, its place is apparently taken by other pro- 
teins of about the same density, and these may be 
the precursors of hemoglobin found in the devel- 
oping cell. There is quite an extensive literature 
showing that "proteins other than hemoglobin" 
are present in the erythrocyte ; one of these is the 
anti-sphering substance, and another is the stro- 
matin of Jorpes, which Boehm believes to be 
anisodiametric and to fill the cell interior. 

2. The classical experiments of "cutting a red 
cell in half", perforating it with glass spicules, and 
some of the modern micrurgical observations lead 
to the conclusion that the cell interior may be 
gelated under certain circumstances. I have even 
gone the length of suggesting that such a gelation 
may contribute to some of the anomalous osmo- 
tic properties (Ponder, 1940). 

3. When ghosts are placed in solutions of 
hemoglobin, there is an adsorption of greater 
quantities of hemoglobin than are likely to be 
bound at the surface (Ponder, 1941, a). The pig- 
ment may be adsorbed on an internal stroma. In 
this connection, it is well known that it is exceed- 
ingly difficult to rid ghosts of the last traces of 
pigment. 

4. The shape of the red cell is not always that 
of a biconcave disc. In some persons the cells are 
oval ("ovalocytosis") like the erythrocytes of 
camels. These ovalocytes show typical disc- 
sphere transformations (Ponder, 1939). In ex- 
perimental and other anemias there appear irreg- 
ularly shaped cells (poikilocytes) which also show 
disc-sphere transformations, but where the dis- 



August 2, 1941 ] 

tortcd parts of the cell apparently "sphere up" 
with difficulty, as if there were some restraint. 
This is very cleaidy seen in the juvenile red cell, 
the reticulocyte, in which bands of material, ap- 
parently situated in the interior, can he stained 
with brilliant cresyl blue ; these persist unchanged 
in form after lysis of the reticulocyte, and appar- 
ently interfere with the disc-sphere transformation 
by binding down the parts of the surface to which 
the bands are attached. 

This last observation, that certain parts of the 
poikilocyte and reticulocyte surface are less mobile 
than others, brings us back to an important ob- 
servation by Furchgott (1940, b). He was able, 
by adjusting the pH of the medium, to make a 
preparation of cells which would become spheres 
as alkali diffused from the glass, but revert to 
discs on the addition of CO2 (by breathing on 
them). In such preparations he observed that the 
biconcavities, and even the larger crenations, ap- 
peared when the sphere turned into the disc at the 
same points at which they were present before the 
disc turned into the sphere. This provides excel- 
lent evidence that the structure of the membrane 
varies from point to point, and W^augh and 
Schmitt's leptoscopic observations (1940) lead to 
the same conclusion. 

4. Conclusion. 

Although it is impossible to set forth all the 
evidence in a review of this length, I think that 
a very fair case can be made out for Norris' theo- 
ry of the shape of the mammalian red cell. It is 
true that he laid rather too much emphasis on the 
liquid nature of the erythrocyte, and that he says 
that "its biconcave shape is due to the operation 



THE COLLECTING NET 



107 



of physical conditions and not to structural re- 
straint", but it is clear that by this he means in- 
ternal structural restraint, for he describes the 
"exquisitely delicate physical pellicle", and it is in 
this that he supposes the physical conditions to 
operate. We would speak of an ultrastructure at 
the surface, and of a special arrangement of mole- 
cules and forces between them. 

A real difficulty, however, remains, and this 
is to explain, on some fundamental grounds (and 
not just by saying "because it was made that 
way"), why the arrangement of molecules and the 
forces between them is such as to give rise to the 
particular biconcave form. This is probably a 
matter for the physicists and the chemists. There 
is, however, another line of investigation to which 
attention is not sufficiently called. The discoidal 
orthochromatic red cell is derived from the larger 
reticulocyte, in which there is a network which 
can be stained with brilliant cresyl blue ; this is 
derived from a nucleated cell, the normoblast, and 
this again from another nucleated cell, from which 
hemoglobin is absent, the pro-erythroblast. The 
discoidal shape seems to be assumed about the 
time when the nucleus of the normoblast breaks 
up and disappears, and we ought not to let the 
accomplishments of physics and spatial chemistry 
make us forget that the investigation of the shape 
of cells still lies in the domain of cytology. Re- 
markably enough, there are few reliable observa- 
tions, and no reliable measurements, on the shape 
of the precursors of the erythrocyte, and this is a 
line of research upon which the biologist can im- 
mediately embark with the expectation of adding 
substantially to our knowledge. 

(This article is based upon a lecture delivered at 
the Marine Biological Laboratory on July 18.) 



SOME ASPECTS OF PIGMENT DEPOSITION IN FEATHER GERMS OF CHICK 

EMBRYOS 

Dr. Ray L. Watterson 

Assistant in Biology, The Johns Hopkins University 



A combined histological and experimental study 
of the developmental history of pigment cells in 
the wing skin and feather germs of Barred Ply- 
mouth Rock embryos has demonstrated that po- 
tential pigment cells, which originate only from 
the neural crest, begin to migrate into the wing 
bud epidermis between 79 and 80 hours of in- 
cubation. Melanoblasts which have successfully 
invaded the epidermis in this way, and their deriv- 
atives by mitotic division, begin the formation of 
pigment between 7 and 71/2 days, thereby becom- 
ing definitive pigment cells ; and by 8 days the 
epidermis of the wing is filled with a network of 
melanophores. Feather germ formation begins 
about this time and is characterized by a thicken- 



ing of the epidermis — first by cell elongation, then 
by cell proliferation — and by an accompanying 
condensation of the underlying dermis, until a 
cap of cells finally protrudes above the surface as 
a definitive feather germ. Any pigment cells 
which are present in the epidermis increase in 
number by cell division and are carried along by 
the niorphogenetic changes in the epidermis. By 
10 days the feather germ is considerably larger 
and is filled with a complex network of pigment 
cells. These much enlarged melanophores appear 
to be distributed entirely at random, and, although 
each is packed with pigment granules, no pigment 
has been deposited as yet. A few hours later this 
random distribution disappears, and all pigment 



108 



THE COLLECTING NET 



[ Vol. XVI, No. 143 



cells in the upper two-thirds of the feather germ 
become grouped into 10 or 11 longitudinal rows. 
In order to understand the nature of this redis- 
tribution of melanophores, it is necessary to ex- 
amine the definitive down feather, and to project 
this structure back upon the feather germ. 

Each down feather consists of 10 to 15 barlis 
held together basally. and each barb bears 2 rows 
of Imrhules. Each barb plus its barbules is desig- 
nated collectively as a liarb-vane. If these com- 
(KMient parts of the l^arb-vane are projected back 
upon the feather germ, they exhibit different spa- 
tial relationships from those seen in the definitive 
feather. Within the feather germ each barb-vane 
is folded up in such a way that it forms a longi- 
tudinal l)arl)-vane ridge jirojecting inward towards 
the ])ulp and hounded peripherally by the feather 
sheath. The liarb lies at the apex, and one row 
of barl)ules occu]5ies each lateral margin of each 
ridge. Since all barb-vanes are folded up in this 
fashion within the feather sheath, the circumfer- 
ence of the feather germ is divided into as many 
longitudinal ridges as there will be barb-vanes in 
the completed feather. It is this breaking up of 
the walls of the feather germ into longitudinal 
ridges which brought aliout the change from the 
random distribution of pigment cells to a definite 
distrilnition into several rows, each row corre- 
sponding to one of these ridges. A somewhat 
earlier stage, characteristic of 11 -day feather 
germs, represents the one time during develop- 
ment that the barbule cells can receive pigment. 
The barl)s have not yet differentiated, and the cell 
bodies of the j^igment cells now lie at the apex of 
each ridge. Their jirocesses extend peripherally 
and carry pigment to the barbule cells. 

It is the nature of this relationship between the 
|)igment cell and the barbule cell which attracts 
our attention. Previously the pigment cell has 
been considered to be almost a micro-injection ap- 
imratus capa]:)le of injecting pigiuent granules 
into passive recipient I)arlnile cells. However, 
several lines of evidence seem to indicate that a 
much more active role is played by barbule cells 
during the pigmentation process. 

(1) Pigment cells are loaded with pigment 
granules at an early stage even before longitudinal 
ridges have formed. Nevertheless, pigment is not 
deposited into any epidermal cells until certain ot 
those cells Ijecome visibly differentiated in the di- 
rection of barbule cells, whereupon they begin to 
receive pigment. 

(2) Before any definite barbule cell differentia- 
tion occurs within a ridge, pigment granules ac- 
cumulate at the tips of pigment cell processes and 
are pinched off and come to lie freely among the 
cells of the ridge. Pigment liberated in this man- 
ner is later taken up by barbule cells. 

(3) Pigment is deposited in each row of bar- 



bule cells in a definite sequence. The melano- 
])hore process extends past the most centrally lo- 
cated barbule cells and first carries pigment to the 
most peripheral cells, and then progressively to 
more axial cells. This is the same order in which 
barbule cells undergo differentiation. The most 
]ieripheral barbule cells are the first to elongate 
and to form keratin, and only when these visible 
dift'erentiation processes begin can they receive 
]iigiuent. As this wave of differentiation spreads 
])rogressively toward the pulp, the more axial 
cells in turn become capable of receiving pigment. 

( 4 ) Pigment cell processes appear to be speci- 
fically attracted toward barbule cells. Under nor- 
mal conditions, none are directed toward cells ly- 
ing between the two rows of barbules. However, 
occasionally a melanophore process does carry 
pigment to a group of cells lying in this axial 
]ilate region. This at first appears to be contra- 
dictory to the idea of a specific attraction, but if 
the fate of these pigmented cells is followed, they 
are found to divide into two groups, and then the 
longitudinal ridge is divided apico-basally between 
them, whereupon these cells which originated in 
the center of one barb-vane ridge become distri- 
buted between two ridges and are distinguishable 
as barbule cells in each ridge. Since normally 
each barb-vane ridge forms one barb-vane, these 
split ridges must form split barb-vanes, which 
they do. Thus, the development of these unusual 
down feathers clearly demonstrates that pigment 
cell processes are specifically attracted to barbule 
cells at this time, even when they must leave their 
usual paths in order to reach them. 

(5) Pigment deposition appears to stinuilate 
the melanophores involved to undergo prolifera- 
tion. If the apico-basal distribution of pigment 
cells which are undergoing mitotic division is 
])lotted from an 11- and a 12-day feather germ, the 
mitotic figures are definitely concentrated within a 
relatively narrow region. It is only within these 
regions that barbule cells are actively receiving 
pigment. 

(6) Finally, a study of the growth curves of 
down feathers reveals an interesting relationship. 
During the first day of its development, the 
feather germ grows slowly, but beginning sudden- 
Iv at 10 days and 18 hours of incubation, the germ 
elongates rapidly, attaining its full growth by 18 
davs. Strikingly, the onset of pigment deposition 
coincides exactly with this onset of rapid growth. 
Lillie and Juhn have estimated that 90% of the 
axial growth of regenerating feathers is accom- 
plished by cell elongation. It would appear that 
])igment deposition begins suddenly at that phase 
of de^•elopment when barbule cells begin to elon- 
gate rapidly. 

The down feather of Barred Rocks is solid 
black in color, whereas the juvenile and adult 



August 2, 1941 ] 



THE COLLECTING NET 



109 



feathers of this breed exhibit alternate black and 
white transverse bars. Willier has demonstrated 
tliat rapidly growing juvenile feathers produce a 
more nearly solid black pattern than more slowly 
growing ones. The down feather grows more 
rapidly than any juvenile feather, elongating 5.5 
mm. between the twelfth and thirteen days, and 
it is indeed a tempting thought that this rapid 
elongation stimulates continuous pigment deposi- 
tion, so that a solid colored feather results. This 
hypothesis is strengthened by a recent observation 
of Mr. James Foulks. If Barred Rock pigment 



cells from a regenerating feather germ, where they 
would normally produce a barred pattern, are 
trans])lanted into feather germs of White Leghorn 
embryos, they deposit a solid black ])attern in the 
host down feathers. 

These several lines of evidence may indicate 
that barbule cells are more important in the pig- 
mentation process than we have previously 
realized. 

(This article is based uoon a seminar report pre- 
sented at the Marine Biological Laboratory on 
July 22.) 



THE INFLUENCE OF HORMONES ON THE DIFFERENTIATION OF MELANO- 

PHORES IN BIRDS 

Dr. Howard L. Hamilton 

Department of Biology. The Johns Hopkins University 



An examination of regenerating feathers from 
birds having red and black in their plumage (e.g.. 
New Hampshire Red fowl, Robin) shows that 
both red and black melanophores are responsible 
for the pigmentation. However, when explants of 
skin from embryos of these birds are grown in a 
tissue culture medium consisting of embryonic ex- 
tract and blood plasma, then black melanophores 
appear, but red ones occur very infrequently in 
the tissue. If sex hormones are added to the cul- 
ture medium, then many red melanophores as well 
as black ones differentiate in the explant. These 
effects on melanophores are similar to those which 
are obtained when hormones are either added or 
removed (by castration) from birds (see Domm, 
'39, for a discussion of plumage color changes). 

Several criteria indicate that red and black 
melanophores are two discrete t3rpes of cells : ( 1 ) 
the pigments are of different colors — orange- 
brown and black, (2) the granules are subspheri- 
cal or pebble-shaped in red melanophores and 
rod-shaped in blacks, (3) red melanin granules 
partially dissolve so that their boundaries become 
blurred when fixed in solutions containing picric 
and acetic acids, whereas black melanin is insolu- 
ble in such reagents, (4) the cytoplasm of red 
melanophores is very fluid as compared with black 
melanophores, because red granules show extreme 
Brownian movement, while black ones move 
slowly due to protoplasmic streaming, and (5) the 
two types of pigment cells react differently to the 
various hormones. 

In general, the sex hormones (estradiol dipro- 
]Monate, estradiol monobenzoate, testosterone pro- 
]iionate, estrone) increase the number of red me- 
lanophores which differentiate in treated explants 
from the New Hampshire Red and Rhode Island 
Red breeds. Sesame and olive oils also produce 
an appreciable stimulation, possibly due to traces 
of sterols. The responses of black melanophores 
from the red breeds to these same hormones are 



more involved. The two esters of estradiol in- 
hibit pigment cells, but estrone and testosterone 
favor their differentiation. 

Because of the similarity of its chemical struc- 
ture to that of testosterone, the adrenal cortical 
hormone, desoxycorticosterone acetate, was used 
on explants from red breeds. The result was a 
nearly complete inhibition of both red and black 
melanophores. A similar reduction in the number 
of melanophores occurred when skin from White 
Leghorn and Barred Plymouth Rock fowl was 
grown in the presence of the cortical hormone. A 
more extensive study on the Barred Rock, which 
possesses only the black type of melanophore, 
showed that sex hormones as well as the cortical 
hormone decrease the number of pigment cells. 
The extent of the inhibition depended somewhat 
on the age of the skin when isolated. Young tis- 
sue (5-6 days) often yielded no melanophores 
when grown in the presence of hormone, but more 
usually (when the hormones were dissolved in 
sesame oil ) there were expanded melanophores, 
but fewer of them than in controls. Skin from 
older embryos (7-8 days) already contains differ- 
entiated melanophores and localized centers where 
feather germs are to form. However, when ex- 
plants are treated with hormones, most of the dif- 
ferentiated melanophores clump and degenerate, 
and only the newly-appearing ones persist as ex- 
panded cells. Furthermore, melanophores aggre- 
gate at loci where feather germs should arise, but 
no structures are formed. This result cannot be 
explained on the basis of a general growth inhibi- 
tion of all cells, because the zone of outgrowth 
from the explant is approximately the same size 
in both the treated and control cultures. When 
crystalline hormones are used there is a reduction 
in number of melanophores, but this is not as 
striking as the delay in their differentiation. The 
red pigment cells in the treated portion of the iso- 
{Continucd on page 112) 



no 



THE COLLECTING NET 



[ Vol. XVL No. 143 



The Collecting Net 

A weekly publication devoted to the scientific work 
at marine biological laboratories. 

Edited by Ware Cattell with the assistance of 
Boris I. Gorokhoff and Judy Woodring. 

Entered as second-class matter, July 11, 1935, at 
the U. S. Post Office at Woods Hole, Massachusetts, 
under the Act of March 3, 1879, and re-entered, 
July 23, 1938. 



CONFERENCE AT THE UNIVERSITY OF 
CHICAGO 

As part of the program celebrating the Semi- 
centennial of the foundinj^ of the University of 
Chicago, a Conference on the Training of Biolo- 
gists will be held. According to an announcement 
of this conference, of which Dr. Paul Weiss is 
chairman, the preparation of a prospective scien- 
tist for his future task, to promote, propagate and 
apply scientific knowledge, requires careful plan- 
ning based on insight into the nature of science 
and its aims, methods, potentialities and limita- 
tions. To discuss the fundamental problems aris- 
ing in the planning of the education of students 
in the Life Sciences, twenty-four scientists and 
educators, representing a variety of disciplines 
bearing on these problems, will gather for a free 
exchange of views and integration of ideas. 

The conference will be held in five sessions, 
rotmd-table fashion, on Thursday, September 18, 
Friday, September 19, and Saturday morning, 
September 20. The general topics are as follows : 

First Session : Introduction — Presentation of 
educational programs in biology currently in op- 
eration in some major universities. Second Ses- 
sion : Contribution to the training of biologists 
from the physical sciences and other related dis- 
ciplines. Third Session : The basic educational 
needs of the biologist. Fourth and Fifth Sessions : 
The specific preparation of biologists for profes- 
sional specialization (research, teaching, medi- 
cine, etc.) 

The morning sessions will start at 9:30 A. M., 
the afternoon sessions at 2:00 P. M. 

LIST OF PARTICIPANTS 
From the University of Chicago: 

Paul A. Weiss, Chairman, Department of Zoology; 
Emmet B. Bay, Department of Medicine; John M. 
Beal, Department of Botany; William Bloom, De- 
partment of Anatomy; Anton J. Carlson, Depart- 
ment of Physiology; Merle Coulter, Department of 
Botany; Earl A. Evans, Jr., Department of Biochem- 
istry; Ralph W. Gerard, Department of Physiology; 
Victor Johnson, Dean of Students, Division of Bio- 
logical Sciences; Wilton M. Krogman, Department of 
Anthropology; George K. K. Link, Department of 
Botany; Carl R. Moore, Department of Zoology; 



William H. Taliaferro, Dean, Division of Biological 
Sciences; Ralph W. Tyler, Department of Education. 
From Other Institutions: 

Detlev W. Bronk, Professor of Biophysics and Di- 
rector of the Eldridge Reeves Johnson Research 
Foundation, University of Pennsylvania; Karl S. 
Lashley, Research Professor of Neuropsychology, 
Harvard University; Dwight E. Minnich, Professor 
and Chairman of the Department of Zoology, Uni- 
versity of Minnesota; Karl P. Schmidt, Chief Cura- 
tor of Zoology, Field Museum of Natural History; 
Francis O. Schmitt, Professor of Biology and Head 
of the Department of Biological Engineering, Mas- 
sachusetts Institute of Technology; Edmund W. Sin- 
nott. Sterling Professor of Botany, Yale University; 
Laurence H. Snyder, Professor of Zoology, Ohio 
State University; C. V. Taylor, Herzstein Professor 
of Biology and Dean of the School of Biological 
Sciences, Stanford University; Benjamin H. Willier, 
Heni-y Walters Professor of Zoology and Chairman 
of the Department of Biology, Johns Hopkins Uni- 
versity. 

DATES OF LEAVING OF INVESTIGATORS 

Baker, Gladys July 21 

Illick, J. T July 23 

Kreezer, G July 25 

Miller, J June 30 

Ronkin, R. R July 25 

Rothstein, A July 14 

Stowell, R. E July 27 

Warner, E. N July 25 

Weaver, Margaret A July 26 

ADDITIONAL INVESTIGATORS 
Benedict, Dora Milton Acad. (Mass.). Br 309. 
Birmingham, L. grad. asst. biol. Rochester. OM 39. 

Dr 3. 
Boyd, M. J. asst. prof, biochem. Cincirmati. Br 341. 
Castelnuovo, Gina zool. Missouri, lib. 
Cole, R. M. grad. fel. biol. Harvard. OM 39. Ka 3. 
Claude. A. assoc. Rockefeller Inst. Br 206. 
Davson, H. assoc. prof. phys. Dalhousie. Br 107. 
Grav, I. E. assoc. prof. zool. Duke. lib. 
Hendricks, Anne L. Cincinnati Med. Br 341. A 202. 
Hopkins, Marjorie G. grad. asst. zool. Mt. Holyoke. 

OM 39. H 3. 
Humm, Frances D. grad. fel. emb. Yale. lib. D 311. 
Keosian, J. asst. prof. biol. Newark. Br 315. 
Morgan, Lilian V. (Pasadena, Calif.). Br 320. 
Muir, R. M. grad. fel. bot. Michigan. Bot. Dr 6. 
Saunders, Grace grad. fel. biol. New York. OM 39. 

H 9. 
Shapiro, H. S. techn. biol. Williams. OM 26. Dr 15. 



CURRENTS 


IN THE HOLE 


At the follo^ving 


hours (Daylight Saving 


Time) the current in the Hole turns to run | 


from Buzzards Bay 


to Vineyard 


Sound: 


Date 


A.M. 


P.M. 


August 2 ,. 


11:57 




August 3 ... 


12:41 


12:57 


August 4 


1:42 


1:57 


August 5 


2:39 


2:53 


August 6 


3:32 


3:46 


August 7 


4:20 


4:35 


Atigust 8 


5:08 


5:23 


August 9 


5:51 


6:09 


In each case the 


current changes approxi- 1 


matelv six hours later and runs from the | 


Sound to the Bay. 




1 



August 2, 1941 ] 



THE COLLECTING NET 



111 



ITEMS OF INTEREST 



Dr. a. J. Waterman, who is instructing in the 
invertebrate course, has been promoted from as- 
sistant to associate professor of biology at Wil- 
Hams College. 

Dr. Henry Emerson, instructor in anatomy at 
the University of Michigan, has been appointed 
instructor in biology at Amherst College. 

Dr. Mary Sears, who is at present working in 
the Woods Hole Oceanographic Institution, has 
received a FacuUy Fellowship at Wellesley, and 
in addition has a grant from the Committee for 
Inter-American Artistic and Cultural Relations to 
conduct research work on Chincha Island, Peru. 
The work will consist of an investigation of plank- 
ton in connection with the feeding habits of Guano 
birds. Beginning August 15 her study will con- 
tinue about eight months, after which time she 
will return to the Oceanographic Institution. 

Dr. Frank A. Brown, Jr., associate professor 
of zoology at Northwestern University, is teach- 
ing a course in comparative physiology and an- 
other in invertebrate hormonal mechanisms at the 
University of Chicago. 

Dr. Samuel A. Mathews, who instructed last 
summer in the invertebrate zoology course, spent 
the month of July at the Scripps Institution of 
Oceanography. 

Dr. Ralph H. Cheney, who has worked at 
the laboratory in previous summers, is on the 
Pacific coast visiting the marine biological sta- 
tions at La Jolla, Corona del Mar, Pacific Grove, 
and Friday Harbor. 

Dr. James A. Miller, who in previous years 
conducted work at the Marine Biological Labora- 
tory, is on the staff of the invertebrate course at 
the Mount Desert Island Biological Laboratory 
this summer. In addition to lectures on the co- 
elenterates, flatworms, and annelids, Dr. Miller 
is making a Kodachrome moving picture of the 
activities of the course. Before leaving the East 
for the University of Michigan, he expects to re- 
turn to Woods Hole for several days. 

Mr. J. Paull, who was registered for the in- 
vertebrate course did not come to Woods Hole ; 
his place has been filled by Miss Louise E. Gross 
of Smith College. 

A new marine biological laboratory is being 
planned by the University of Texas. Located on 
the Texas coast of the Gulf of Mexico, the Lab- 
oratory has been granted $25,000 by the General 
Education Board. Additional funds will be neces- 
sary, however, before construction can be started. 

The Atlantis returned from dry dock at the end 
of this week, and will leave Sunday or Monday 
for the other side of the Gulf Stream. 



Dr. and Mrs. Charles O. Warren visited 
Woods Hole over the weekend. Both Dr. War- 
ren and his wife, the former Katherine S. Brehme, 
have worked at Woods Hole in the past. They 
are spending the summer at the Biological Lab- 
oratory at Cold Spring Harbor. 

Miss Annabelle Broomall was married to 
Dr. Richard Horn on June 11 in Wilkinsburg, 
Pennsylvania. She is working at the Laboratory 
on the genetics of a parasitic wasp, and this fall 
will work for her doctor's degree at the Univer- 
sity of Pittsburgh. Her husband is now interning 
in the Pittsburgh Medical Center. 

At the weekly staff meeting at the Woods Hole 
Oceanographic Institution on Thursday evening. 
Dr. Theodor van Brand, of the Catholic Univer- 
sity of America, spoke on "Experimental Studies 
upon the Nitrogen Cycle in the Sea." 

Motion pictures on "Seals in Alaska" were 
shown Thursday evening in the M.B.L. Auditor- 
ium. The film was provided by the Fish and 
Wildlife Service and explanatory comments were 
made by Dr. P. S. Galtsoff. 

Mr. Lower's Movies of Aquarium Life and 
Children this summer will be shown on Friday, 
August 8, 3 :00 P. M. in the Schoolhouse for the 
benefit of the microscope fund. 20c children, 40c 
adults. The Annual Exhibition of Children's 
Work will take place from 2 to 5 P. M., Friday, 
August 8. 

The program of the phonograph record concert 
at the M.B.L. Club Monday night is as follows: 
Beethoven, "Egmont Overture;" Prokoffiev, "Pe- 
ter and the Wolf;" intermission; Tschaikowsky, 
"Symphony No. 4." 

M.B.L. TENNIS CLUB 

The officers of the Tennis Club will appoint a 
committee to run a tournament of events in ladies' 
and men's singles and doubles, provided enough 
players care to enter. The entries must be in by 
Monday, August 4, and drawings will be made 
that evening. Play will begin immediately in or- 
der to finish the finals by the middle of August. 
Entries should be made on the sheets posted by 
the Mess court. 

The courts are less crowded than usual this 
summer, and the club needs the support of the 
community to reduce its indebtedness. The havoc 
caused to the new beach courts by the tidal wave 
in 1938 brought about heavy expenditures for re- 
construction. During the past two seasons several 
club members have saved the club about two hun- 
dred dollars by putting the surface into playing 
condition. In addition, the club is grateful to 
several people who have given generous aid and 
so helped to reduce the club debt. — D. E. L. 



112 



THE COLLECTING NET 



[ Vol. XVI, No. 143 



STUDIES ON THE LIFE HISTORY OF SIPHODERA VINALEDWARDSII, 
A TREMATODE PARASITE OF THE TOADFISH 

Dr.s. R. M. Cable .\n'd .\. V. Hunninen 
Purdue L'lin'crsitv aud Oklahoma City Uiik'crsity 



Experimental studies on the life history of 
Siphodcra viualcdtvardsii (Linton) have demon- 
strated that this trematode is related to the Heter- 
o]ihyidae as ]5ostulated by Manter. Price, and 
\\'ilhelnii on the basis of morphological and sero- 
logical investigations. The definitive host in the 
W^oods Hole region is the toadfish. Opsanus fau. 
jjractically all of which are infected. The small 
marine snail, Blttiuiu altcrnatuiu, serves as the 
molluscan host in which the cercariae develop in 
sim]5le, elongate rediae. The cercaria is a pleuro- 
lojihocercous form of an unusual type since the 
tail is inserted \'entrally and coiled when at rest, 
the fourteen jienetration glands have two instead 
of the usual four bundles of ducts in the region 
of the oral sucker, and the excretorv formula is 



2 [(2+2) -f (2+2)] = 16 flame cells. The cer- 
cariae penetrate and encyst in various species of 
flounders, developing into infective metacercariae 
in ap]5roximately two weeks. Metacercariae occur 
in the fins, body wall and even the myocardium of 
the fish. Feeding experiments thus far completed 
indicate that toadfish become infected by eating 
fish containing metacercariae. Three toadfish, 
isolated for four weeks, were fed fish containing 
1 3-day metacercariae. Two of these have been 
examined and found to contain large numbers of 
\ery young worms in addition to a few mature 
specimens from previous natural infection. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on 
•July 15.) 



THE INFLUENCE OF HORMONES ON THE DIFFERENTIATION OF MELANO- 

PHORES IN BIRDS 

(Continued from page 109) 



late contain fewer melanin granules, so that they 
appear lighter-colored, and granules are not elim- 
inated as soon from theuL Though chronologi- 
cally of the same age, these treated melanophores 
are physiologically younger than the controls. 

In summary, it is seen that genetic differences 
in the precursor cells must determine whether 
thev become red or black melanophores, since ]:)oth 
kinds develo]) adjacent to one another in the same 
piece of tissue and in the same clot. However, 
certain environmental factors must influence 
which of the two kinds of melanophores will pre- 
dominate. For e.xam]-)le, fast-growing feathers 
are mostly black in the Rhode Island Red breed, 
so that physiological differences in feather germs 
must favor the clift'erentiation of one or the other 
kind of pigment cell. This study also shows that 
hormones (and probably other sterols) increase 
or decrease the numbers of each kind of melano- 
phore. Hormones apparently act directly on me- 
lanophores, since they are fairly well-isolated /')) 
vitro from other cell types which might affect 
them. The evidence suggests that the hormone 
exerts some intracellular metabolic effect so as 
either to catalyze or inhibit melanin synthesis. Ap- 
parently there are no transitional stages between 
red and black melanophores ; hormones do not 
cause one kind of pigment cell to change into an- 
other. Red melanophores are not produced b)' 



sto]iping normal melanin synthesis at a red stage. 
If this were true, all black melanophores should 
]mss through a red phase in their formation, and 
this is not observed. Hormones cause the appear- 
ance of red melanophores only in those birds 
which have genes for red pigment (i.e., possess 
red-type precursor cells ) . These precursors or 
melanoblasts remain latent in the tissue until con- 
ditions are favorable, either in cultures or in 
feather germs, for them to synthesize melanin. 

.\n examination of human hair follicles shows 
that both red and black melanophores are present 
in some individuals, and that different proportions 
of these two kinds of pigment cells account for the 
\-arying shades of brown, sandy, and red hair. 
Dark brown and black hairs contain only black 
melanophores. The similarity of these two kinds 
of mammalian jjignient cells to those of birds sug- 
gests that they, too, might be influenced by hor- 
mones. Graying in hair and whiteness in feathers 
are similar in that both are caused by loss of pig- 
ment cells (due to decreased viability of melano- 
]ihores or adverse environmental conditions in the 
follicle) so that those few which remain are un- 
able to produce enough pigment to color the hair 
or feather. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on 
July 22.) 



August 2. 1941 ] 



THE COLLECTING NET 



113 



INVERTEBRATE CLASS NOTES 



We invertebrates, lifty-five in all. representinj:;- 
forty-eight colleges and universities, had our first 
meeting Thursday evening, July 24. At this time, 
jovial Dr. Bissonnette introduced our instructors, 
explaining the workings of the course, warned us 
about the tides, poison ivy. and making sea water ■ 
do the work of formalin. 

Protozoans were the sul^ject of three lectures 
liy Dr. Waterman, in which he not only reviewed 
the structural and ecological aspects, but also some 
of the important research problems of this group. 
For us beginners at least, the lab work became 
more difficult as we turned from attached forms 
to parasites and free-swimming forms. The lab 
was by no means deserted over Sunday, some of 
us striving to cover a respectable minimum, others 
to take advantage of the abundant material. In- 
deed, we have become quite familiar with the 
customary peroration : "There is sufficient mater- 
ial for a week's work. Please hand in your lali 
records by tomorrow noon." 

Dr. Lucas had the tough assignment of crowd- 
ing into a single afternoon a comprehensive lec- 
ture on the somewhat anomalous Porifera. to 1)e 
followed by our investigations of the several 
types available. To carry out our end of the job. 
many of us worked well into the night. The fail- 
ure of the power supply at one A. M. sent the 
last investigators groping to bed. 

Spicules and choanocytes duly observed and re- 
corded, we have turned our attention to polyps 
and medusae. Guided by Dr. Crowell's well or- 
ganized lectures we have been examining the di- 
verse types and phases of hydroid life. 

Our first two days were further enlivened by 
field trips, one to Stoney Beach, the other to 



Lackey's Bay. Equipped with the "Ark" — fully 
as capable as its Biblical prototype at taking, two 
by two or as fast as we find them, any and all 
marine invertebrates — watch glasses, forceps, hand 
lenses, we readily found representatives of most 
(jf the invertebrate phyla. Identification was an- 
other matter, one largely in the hands of our om- 
niscient instructors. By the second trip however, 
the "recording angel" was hard put to keep track 
of the staccato calls of "Amaroucium", "Nassa 
trevetata," "Diapatra," scarcely audible above the 
other sounds associated with a successful field 
trip. 

As we're a social noisy group, we received the 
news of the M.B.L. Mixer with enthusiasm. Few 
of us went with the sole purpose of meeting cele- 
brated investigators, but the rest of us preferred 
the frivolous chatter and dancing. With our back- 
ground in Protozoology we added an appropriate 
(purely scientific, of course) finish to the day by 
swimming in the biluminous sea. Several active 
souls finished the evening in the dab. boiling un- 
wanted lobster claws and playing bridge. 

During lab hours (i.e., any time) we hear the 
results of John Osmun's newl}' (quite newly) 
formed quartet with Bob Porter, Howy Miner, 
and Sid Pond. The one person that will eiifec- 
tively interrupt them is Gary Metcalf of the col- 
lecting crew who never fails to praise the crew's 
soft l:)all team. We have met the challenge by or- 
ganizing a volunteer eleven (not to mention a 
complete girl's team). Dr. Rankin has promised 
to be our first base man. This stroke of luck 
should remove all doubt concerning the final 
scores. A merciful rain postponed our first game. 
— Louise Gross and Bill Batchclor 



EFFECT OF SEA WATER ON THE RADIOSENSITIVITY OF ARBACIA SPERM 

(Continued from page 101) 

Table I. 
Percentage Fertilization 



Amount of radiation: 
76,500 roentgens. 



Irradiated 



Controls 



Cone, of sperm : "Dry" 1:5 1:10 1:100 "Dry" 1:5 1:10 1:100 

Tinie of insemi- 
nation after 
irradiation : 

60 min. : 100 100 87 3 100 100 100 100 

90min. : 98 62 19 2 99 100 91 98 



from the alcove data that another indication of in- 
creased sensitivity at greater dilutions is a de- 
crease in the ability to survive over a period of 
time. It appears that some sperm, not killed in- 
stantaneously, may be injured so that they die 
later. 



Because of the iJossibility that the increase in 
sensitivity might be due to production of toxic 
materials in the water, irradiated sea water was 
added to sperm but no definite injury was ob- 
served. 

A quantitative study of the change in resistance 



114 




THE COLLECTING NET 




[ Vol. XVI, No. 143 






Table II. 






Amount of 


radiation : 


None 1.160 1- 2.120 r 


3.180 r 


4.240 r 5.300 r 



Percent, fert. : 

Cone. .Sperm 1:2000 
Percent, fert. : 



100 


69.5 


45.0 


35.4 


41.5 


33.8 


100 


7.0 


7.1 


3.5 


13.2 


7.95 



Amomit of radiation : 



None 1,195 r 2.390 r 4.780 r 9,560 r 19,120 r 



Cone, of Sperm 1:10 
Percent, fert. : 

Cone. .s])erm 1 : 1000 
Percent, fert. : 



100 


100 


100 


100 


93.0 


93.0 


100 


73.0 


63.8 


36.6 


7.1 


1.77 



Amount of radiation : 



None 1,190 r 2,380 r 4,760 r 9,520 r 19,040 r 



Cone, of Sperm 1:100 
Percent, fert. : 

Cone, of Sperm 1:1000 
Percent, fert. : 



100 


97.2 


98.3 


99.3 


92.1 


56.9 


100 


94.7 


94.4 


21.7 


6.7 






Amount of radiation : 



None 5.600 r 1 1.200 r 22,400 r 44,800 r 89,600 r 



Cone. Sperm 1 :20 
Percent, fert. : 

Cone. Sperm 1 : 200 
Perct. fert. : 



100 


86.0 


86.0 


68.5 


48.8 


40.7 


100 


12.6 


1.5 












of the s|Derm at various concentrations was made 
in the followin"; manner. Sperm were collected 
in as concentrated a form as possible, such sperm 
lieing considered as "dry" or 100% sperm. The 
desired dilutions were made from the same lot of 
"dry" sperm, and, after 15 minutes, were irra- 
diated. Following irradiation, fertilization tests 
were made. Each lot of sjierm was diluted to the 
same concentration before insemination in order 
that the same amoiuit of sperm would be used in 
each case for the same number of eggs. The 
amount of sperm added to the eggs was not suffi- 
cient to give 100% fertilization. In this manner, 
an excess of sperm was avoided and small changes 
in ability to fertilize could be detected. At least 
two lots of eggs were fertilized in each case and 
several counts of each were made. The final 
counts were made at the time when most of the 
fertilized eggs were in the two cell stage. The 
results were expressed as percentage fertilization 
(as compared to controls). .As shown in Table 
II, the ability to fertilize is markedly altered by 
the proportion of water (present during the irra- 
diation. It should be noted that the dilution ex- 
periments were performed in pairs in order to 
prevent possible dififerences due to variation in 
radiosensitivity from one lot of sperm to another. 
Therefore the values to be compared are those 



olrtained by different dilutions in the same experi- 
ment and not necessarily from one experiment to 
another. However, the results of all of these ex- 
periments are in general agreement. 

Some exjieriments have been performed in an 
effort to obtain some information regarding the 
mechanisms controlling the change in radiosensi- 
tivity. It has been found that dilute sperm sus- 
pensions irradiated immediately after the addition 
of water { when the rate of oxygen consumption 
is high) are more susceptible than sperm that 
have lieen in sea water for 30 minutes (at which 
time the rate of oxygen consumption has dropped 
to a lower level). This, together with the fact 
that the addition of sea water greatly increases 
the activity and rate of oxygen consumption, in- 
dicates that these factors play a part in determin- 
ing the radiosensitivity of the sperm. Other fac- 
tors are being investigated in an attempt to eluci- 
date the mode of action of radiation on the fertil- 
ity of sperm. 

Preliminary experiments on Nereis sperm in- 
dicate a similar relation between dilution with sea 
water and radiosensitivity. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on 
July 29.) 



August 2, 1941 ] 



THE COLLECTING NET 



115 




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49 



116 



THE COLLECTING NET 



[ Vol. XVI, No. 143 



DeGa 



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THE COLLECTING NET 



117 



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Authors of biology texts and manuals fre- 
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118 



THE COLLECTING NET 



[ Vol. XVI, No. 143 



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THE COLLECTING NET 



119 




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120: 



THE COLLECTING NET 



[ Vol. XVL No. 143 




IMiisemn of Natural History, New York 



When Gorilla j"/7zy/^^/ Visits a City Classroom 



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Vol. XVI, No. 7 



SATURDAY, AUGUST 9, 1941 



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EFFECT OF AZIDE ON CYPRIDINA 
LUCIFERIN 

Dr. Aurin M. Chase 
Physiological Laboratory, Princeton University 

Luciferin is the substrate in the bioluminescent 
reaction. In the presence of an enzyme, hicif er- 
ase, and of oxygen in solution, light is produced. 

Reversible oxidation of luci- 

ferin can occur in the absence 
of luciferase but there is no 
luminescence in this case. (E. 
N. Harvey, Ann. Rev. Bio- 
chcm.. 1941.) 

The luciferin and luciferase 
used in the present experi- 
ments were obtained from the 
crustacean, Cypridina hilgen- 
dorfii. In the thoroughly dried 
organisms the luminescent ma- 
terials remain stable practical- 
ly indefinitely. The luciferin 
was extracted and ])urified by 
the method of Anderson (/. 
Gen. Physiol., 1935) and the 
luciferase was partially puri- 
fied b y prolonged dialysis 
against distilled water. The 

i-eaction mixtures, of pH 5.4 

and 6.6, were made up from 
phosphate buffer, 0.2 M. and the sodium azide 
solutions were also made up in these same buffers. 
Luminescence was (Continued on page 131 ) 



M' % % (Ealenbar 



TUESDAY. August 12, 8:00 P. M. 

Seminar: Dr. W. Trager: "Studies 
on Conditions Affecting the Sur- 
vival in vitro of a Malarial Para- 
site." 

Dr. Charles Hassett: "The Effect 
of Dyes on the Response to Light 
in Peranema." 

Dr. J. 0. Hutchens: "Utilization of 
Ammonia by Chilomonas Para- 
mecium." 

Miss Virginia Dewey and Dr. G. 
W. Kidder: "The Possibility of 
Thiamine Synthesis by Ciliates." 



FRIDAY, August 1.5, 8:00 P. M. 
Lecture: Dr. Franz Schrader: "The 
Mitotic Movements of Chromo- 
somes." 



PROTEINS IN ACTION 

Dr. Dorothy Wrincii: 

Professor at Amherst, Smith and 

Mount Hoi yoke Colleges 

One of the most interesting characteristics of 
this golden age of biological research is the way 
in \\-hich a new synthesis of ideas is emerging, 

as the result of the immense 

])rogress which has been 
achieved by specialist investi- 
gators working in a score of 
highly technical biological 
fields and the rise of the new 
science of structure chemistry. 
The original Imrriers between 
morphological and functional 
studies are giving way before 
the realization of the fact that 
function can only be under- 
stood in terms of structure, so 
that structure, far from being 
the concern onl)' of certain 
specialists, is seen to be of the 
first importance in a great 
variety of attacks on biological 
problems. Biological systems 
indeed are now seen to be 
superlatively fine examples of 
functional architecture. Simul- 
taneously structure chemistry, the science which 
studies the geometry of intra- and inter-molecular 
atomic arrangements, has come of age. This 



TABLE OF CONTENTS 



Proteins in Action, Dr. Dorothy Wrinch 121 

Effect of Azide on Cypridina Luciferin, Dr. 
A. M. Chase 121 

Nominations for Trustees 128 

Representation by Institutions at the M. B. L. 128 

Additional Investigators 128 



Dates of Leaving of Investigators 128 

Items of Interest 129 

Invertebrate Class Notes 130 

Review of "Embryology of Insects and My- 

riapods," Peter Gray 130 




ALONG WATER STREET DURING THE 1938 HURRICANE 

Above: The M.B.L. Club House at the height of the storm, with the Woods Hole Oceanographicj 
Institution behind it. The building between the two telephone poles at the left is Rowe's Drug Store,] 
while that to the right of the Oceanographic building is the Institution's pump house. 

Below: A churning mass of wreckage being tossed over the sea wall, photographed from the steps 
of the Yalden Sundail. At the extreme right is the M.B.L. pump house. On the horizon may be dis- 
tinguished Lawyer Hathaway's yacht "Rosemay," which battled the storm successfully by running its 
engines at full speed throughout the hurricane. (Photographs by L. A. Baker) 



August 9, 1941 ] 



THE COLLECTING NET 



123 



new science, with which the names of Lang'- 
muir, Lewis, and Pauling in this country, and 
W. H. Bragg and W. L. Bragg in England 
will always be associated, has already accom- 
plished for many inorganic materials, such as 
diamond, graphite, the silicates, and water, 
among countless others, just the type of analysis 
which is required for the deeper understanding of 
biological materials. It has shown how mechani- 
cal and physical properties can be explained in 
terms of atomic arrangements. The hardness of 
diamond and the flakiness of graphite, the softness 
of naphthalene, the water-holding capacity of the 
clay minerals, the variation in properties in the 
different classes of silicates, for example the platey 
character of the micas and the fibrous nature of 
the amphiboles — all can be and indeed have been 
seen to be direct consequences of atomic patterns. 
The new synthesis in biology now developing has 
as part of its objective the strictly similar task of 
the analysis and interpretation of biological data 
in atomic and molecular terms. In innumerable 
ways the triumphant transition from millimeters 
to Angstroms of the structure chemist working 
with inorganic materials and the simpler organic 
substances offers pointers to the biologist so that 
he may in due time accomplish a similar transi- 
tion, the narrower transition from the micron, 
even perhaps (with the help of the electron micro- 
scope) the decimicron, to the Angstrom world. 

Tonight I shall ask you to review with me re- 
cent findings in a number of experimental fields 
which can be interpreted in terms of structure 
chemistry and to consider a number of specific 
suggestions regarding the possible nature of bio- 
logical structure systems which result. 

Cytoplasm in Focus 

Structure problems in biology find a focus in 
the nature of cytoplasm in general including par- 
ticularly the biologically functioning membranes, 
the cell plasma membranes around all animal and 
plant cells, the nuclear membrane and the mem- 
brane of red cells. A great deal is already known 
as to the mechanical and physical properties of 
these systems, something also of their chemical 
nature. Perhaps the most striking data of all are 
those which indicate an inherent dichotomy in the 
nature of cytoplasm in general. Thus if, because 
of its high water content, we wish to consider 
cytoplasm as a liquid system, we must admit, as 
Frey-Wyssling and others have pointed out, it is 
a very anomalous one, since it does not satisfy the 
usual criteria. It is not inelastic, so it is a non- 



Newtonian liquid. It does not have a constant 
viscosity at a given temperature, so it is a non- 
Poiseuillian liquid. Small particles in cytoplasm 
do not rise or fall with constant velocity, so it is 
a non-Stokesian liquid. It is not under all cir- 
cumstances optically isotropic, so in a fourth re- 
spect it forfeits its title to being considered an 
ideal liquid. Furthermore, cytoplasm has a cer- 
tain rigidity ; it evinces under appropriate circum- 
stances a certain constancy of shape ; it possesses 
some tensile strength. 

Any attempt to think of cytoplasm in terms of 
ordinary ideas of solid structures is equally un- 
successful. Cytoplasm is extremely deformable 
and very flexible. It is characterized also by the 
power to shrink and swell reversibly on the grand 
scale, its essential structure, upon which its func- 
tional behavior depends, apparently remaining un- 
disturbed. It has a huge water content which ap- 
proaches even 97 per cent in the case of certain 
cells and the water easily allows the passage of 
certain substances, though not of others. 

This fact draws attention to the most character- 
istic property of all, namely the extreme specificity 
of the cytoplasm as evinced by its permeability 
properties in general and in other ways, a fact 
which indicates the necessity for a totally different 
type of structure from that of classical chemical 
systems, whether thought of as solids or liquids. 
The central points about cytoplasm were put in 
the clearest manner by Wilson many years ago. 
"Nothing is more remarkable," said Wilson writ- 
ing several decades ago, "than that a thing so 
delicate and plastic should run so true to form 
through countless generations. How this is pos- 
sible we can hardly imagine. We can but re- 
cord the observed fact that it is effected by an in- 
herent power of adjustment and self-regulation 
which holds the cell fast to its own type." What 
then is the nature of this structure by means of 
which, as Wilson says, "within certain well-de- 
fined limits the activities of every cell are of spe- 
cific type," such that "the muscle cell, the nerve 
cell or the gland cell displays its own characteris- 
tic performance" ? 

Proteins, the Clue 

The clue to the mystery is not far to seek. It 
resides in this fact : no consideration of any struc- 
ture problem in biology or medicine can be com- 
plete unless due account is taken of the protein 
constituent. I wish specially to make clear that 
no one with any appreciation of the recent ad- 
vances in knowledge of the parts played by many 



The Collecting Net was entered as second-class matter July 11, 1935, at the Post Office at Woods Hole, Mass., 
under the Act of March 3, 1879, and was re-entered on July 23, 1938. It is devoted to the seientifie work at 
marine biological laboratories. It is published weekly for ten weeks between July 1 and September 15 from Woods 

Hole, and is printed at The Darwin Press, New Bedford, Mass. ""- -^=^-=-- -«= ;i„„<.„j :„ to„„.:i„ ti„i„ 

Mass. Single copies, 30c by mail; subscription, $2.00. 



Its editorial offices are situated in Woods Hole, 



124 



THE COLLECTING NET 



[ Vol. XVI, No. 144 



non-protein components in living systems could 
be willing to assert that the protein alone is the 
whole story. We have only to think of the im- 
portance of the lipoid component in red cell mem- 
branes and of the carbohydrates, as for example 
in the pneumococcal polysaccharides, to see the 
necessity for this qualification. This aspect how- 
ever has not lacked emphasis. Since the turn of 
the century there has been a tendency to ignore 
proteins in physiological problems because exact 
studies on simpler substances of biological impor- 
tance have naturally taken precedence. In conse- 
quence there has been, in some quarters at least, 
a tardiness to assess at its real value the role of 
proteins in biological structure problems. 

Thus enzymes were denied protein status as 
late as 1926 by no less an authority than Will- 
statter. Curiously enough, in this same year 
urease was crystallized by Sumner at Ithaca and 
insulin was crystallized by Abel at Hopkins. 
Proof that many enzymes are proteins has since 
been given by Northrop and others. Certain cel- 
lular respiratory enz)^mes which operate through a 
prosthetic group depend for their specificity on the 
protein portion of the molecule. Further, several 
vitamins have l^een shown to be prosthetic groups 
which associate with proteins. 

Similarly, it has recently appeared that most 
hormones either are proteins, e.g.. insulin, the 
parathyroid and pituitary hormones, or else exist 
in the body in combination with a protein, e.g., 
thyroglobuiin. The sterol hormones are an im- 
portant group lor which evidence of protein com- 
bination is lacking, luit it is interesting to note 
recent evidence for a combination between choles- 
terol and pneumococcal hemolysin. 

The startling demonstration that the viruses are 
complex nucleo-proteins has also been a develop- 
ment of the last few years. The remarkable prop- 
erty of the viruses of being able to enter cells and 
increase in concentration, presumably by an auto- 
catalytic process, puts these molecules on the 
border line between the living and the inanimate. 
It may be permissible (as Muller, for example, 
has maintained) to picture the viruses as the pro- 
totype of the genie material itself, which is able 
to go further and elaborate a cellular environment. 
These developments mean that the time is ripe for 
a new appraisal of the significance of proteins in 
biology and medicine. Proteins can no longer be 
regarded as relatively inert materials "which exert 
a colloid-osmotic pressure" : they are emerging as 
the key substances in physiological processes. 
They are the key even in the sense that many 
other biologically-active molecules may owe their 
significance to their relationship to the proteins. 
It is my opinion that the realization of all the im- 
plications of these facts will mark the beginning 
of a new era in the biological sciences. 



Structure Forming Proteins 

But the question then arises as to how proteins 
play this trancendent role in biological systems. 
The answer has been stated with admirable preci- 
sion by R. S. Lillie over twenty years ago when 
he surmised that "the specific characteristics of 
every animal or plant may be determined ulti- 
mately In' their structure-forming proteins." We 
cannot doubt that proteins are the structural 
units which, with some other subsidiary materials, 
build the structures, which the histologists meas- 
ure in microns, out of the ordered atomic patterns, 
which chemists measure in Angstroms. There 
can be little doubt that proteins, in their structure- 
forming capacity, are the main components of the 
conductile and contractile mechanisms, and of the 
cell machinery involved in the process of secre- 
tion and absorption. An understanding of the 
processes involved in norma! growth and the 
healing of wounds naturally requires a knowledge 
of the mode of synthesis of tissue constituents, 
and therefore also of protein synthesis. 

The recent developments in our knowledge of 
enzymes, hormones, viruses and genes to which 
I have already referred are indeed a striking 
fulfillment of the judgment of a philosopher of the 
last century who said, already in 1878: "Life is 
the mode of existence of protein sulDStances." 
Just how minutely, recent accessions of knowledge 
of what we may call "the biology of the proteins" 
allow us to work out the picture of the proteins 
as the essential "structure-forming" components 
of living systems. I shall now try to show, even 
if only in the barest outline. 

The main sources of our information are the 
])hysical chemistry of proteins, studies of anti- 
bodies and the crystallography of native proteins. 
W'hile these are by no means the only sources of 
information relevant to our investigation of the 
"structure-forming proteins." I would emphasize 
one important aspect. All these techniques have 
proved themselves capable of yielding information 
about native proteins. In the attempt to under- 
stand the nature of such biologically active sys- 
tems as cytoplasm and plasma membranes, im- 
mense emphasis in recent years has been laid on 
the surmises and suggestions contained in early 
work on dead proteins such as wool, silk and 
gelatin, some of which have recently been retract- 
ed by the leading worker in the field, W. T. Ast- 
bury. Even if the whole subject of the structure 
of these inert proteins were not in a complete state 
of flux, there is no reason to think that this is a 
fruitful source of inspiration for those concerned 
with living systems, since these dead proteins have 
none of the properties which physiologists and 
biologists require of actively functioning proteins. 
It seems to be little realized how far developed is 
the present day picture of the native protein, and 



August 9, 1941 ] 



THE COLLECTING NET 



125 



how fundamentally it differs from what is known 
about the grosser properties of dead proteins and 
from what has been sugg^ested, admittedly on the 
basis of very inadequate X-ray and chemical data, 
for the atomic structure. I understand from the 
Director that it is not necessary that I try to hide 
from you the fact that, in my opinion, there is a 
very real crisis in this field of protein structure 
today. The tradition regarding these Evening 
Lectures was in fact laid down in 1893. "Some- 
what greater freedom in the expression of opinion 
than might be expected in strictly scientific com- 
munications must be permitted," writes the first 
Director of the Laboratory in the preface of the 
first of the familiar blue volumes of "Biological 
Lectures." "In fact it is one of the leading ob- 
jects of these Evening Lectures to bring forward 
the unsettled problems of the day and to discuss 
them freely." Discussing, then, this crisis freely, 
I would offer it, as m)' opinion, that, of all the 
elements in the complex protein field today which 
are holding up the progress of the understanding 
of structure problems relating to living matter, the 
one most responsible for the present deadlock is 
the hypothesis that polypeptide chains are the es- 
sential constituents of native proteins. The recent 
developments I mentioned make it quite possible 
that the picture of the structure of hair and the 
rest may need the most thoroughgoing revision, 
far beyond the modifications recently suggested in 
Nature. These modifications replace one naive 
picture based upon insufficient evidence by an- 
other, hardly less naive, and equally ill-ground- 
ed in fact. Be the situation in this field as 
it may, there seems to me to be no question 
but that the polypeptide chain hypothesis of 
hundreds of residues handcuffed in pairs is not 
only unproven but extremely mischievous, mis- 
chievous in its inhibiting effect upon attempts at 
protein synthesis now long overdue, mischievous 
also in the influence on the design of experiments 
in more than one field of physiology. Pressed to 
define where they stand on the polypeptide chain 

, theory, there prove to be few protagonists of this 
simple picture who will not admit that the chains 
(it present) must be interlinked in definite pat- 
terns. Yet the fact that this admission leads di- 
rectly to two or three dimensional patterns of the 
characteristic residues and all the far reaching 
implications of this obvious idea remain almost 
universally unappreciated. 

Tonight, drawing the most definite of lines be- 
tween dead and dying proteins and biologically- 

j active native proteins, I propose to assume that 

j the proteins in cytoplasmic and membrane struc- 
tures are in the latter form. It ma}' not be pos- 
sible, in the present crisis, to make out a water- 
tight case for the assumption, but it seems diffi- 
cult indeed to see where, in living organisms, they 

I would be in the native state if not here. It ap- 



pears, however, legitimate for the unprejudiced 
observer to read a striking confirmation of the as- 
sumption into the recent work of Ballentine and 
Parpart who found that erythrocytes subjected to 
crystalline trypsin at pH 6.8 showed no change 
in permeability even after 24 hours exposure to 
the enzyme, a result which mcidentally surely 
should cause acute embarrassment to polypeptide 
chain supporters. 

The Plain Unvarnished Facts 

If then we wish to understand the present day 
picture of the native protein, let us first consider 
the results obtained lay Svedberg with his ultra 
centrifuge. This work opened a new era in the 
understanding of proteins, for contrary to the ex- 
pectations of many whose techniques consisted in 
first tearing to pieces the delicate structures of 
these biologically-active substances and then study- 
ing the resulting chaos, he found that native pro- 
teins are made up of well-defined molecular 
species. Of course the air-tight proof of molecu- 
lar status for any one protein involves investiga- 
tions of the highest dehcacy, including above all 
solubility studies. But by and large the picture 
of protein units as genuine megamolecules emerges 
unmistakably from his researches. There also 
emerges a second characteristic of proteins, name- 
ly the dependence of their stability upon interlink- 
ing or association with other molecules or ions. 
The other molecules always, so far as we know, 
include water ; many other types of molecules, in- 
cluding other proteins, also play a part. This 
seems to be the meaning of the phenomenon of re- 
versible dissociation of native proteins, first uncov- 
ered by Svedberg's researches, of which there are 
many examples, including horse hemoglobin, the 
seed globulins, the hemocyanins and the erythro- 
cruorins. Such dissociable proteins (which fall, 
it is claimed into aliquot parts), break up into 
smaller units which still have the protein charac- 
ter. They are therefore more properly called pro- 
tein particles. That the stability of a protein unit 
or of a protein particle or colony of protein units 
can be ensured by linking of protein units together 
instead of the linking of each with a full comple- 
ment of waters or other "foreign" molecules is 
shown in such a case as horse hemoglobin, where 
plain dilution with water causes the presumably 
dimeric molecule present when the concentration 
is as high as one per cent to be replaced by (half- 
sized) monomeric molecules when the concentra- 
tion is below 0.5 per cent. Dissociations of other 
proteins by dilution with many other molecular 
species of polar molecules, including arginine, ly- 
sine and so on are also known to occur. 

The picture of the proteins as genuine mole- 
cules, whether monomeric or polymeric, which 
comes from these physico-chemical studies is amp- 
ly confirmed by crystallographic studies which 



126 



THE COLLECTING NET 



[ Vol. XVI, No. 144 



paint the same picture, with even greater sureness. 
These studies have shown that native protein 
molecules have definite atomic patterns. The X- 
ray pictures of horse hemoglobin are of such per- 
fection that there can be no doubt that each mole- 
cule consists of a highly organized framework of 
atoms. Further, the transition from the "wet" to 
the "dry" crystals of this and other proteins indi- 
cates that some or all the R-groups of the resi- 
dues, although rooted in definite spatial patterns 
on or in skeleton frameworks (whether of surface 
or volume type ) , are capable of considerable elas- 
ticity and flexibility. 

The immunological picture is entirely in accord. 
Marrack, in fact, goes so far as to picture certain 
constellations of R-groups in definite spatial ar- 
rangements dotted about on the rigid surface of 
the native protein as the most natural interpreta- 
tion of the imnnmological facts. How otherwi.se. 
indeed, can we interpret the subtle differences in 
the affinities of antibodies formed by an antigen 
formed from l-phenyl-2, 3-dimethyl-4-amino- 
5-pyrazolon diazotized and coupled with protein 
and that formed when the haptene changes the 
position of a double bond. 

The deduction by Alarrack that surfaces of pro- 
teins carry R-groups in definite patterns leads us 
to another aspect of the matter which is perhaps 
one of the most crucial of all for biological struc- 
ture problems. Evidently we have to assume that 
the patterns on one patch of a native protein, or 
shall we not rather say one face ( since the sur- 
face as a whole must have a definite morphology 
which e^•en in the most general case must be poly- 
hedral in type) fits with certain faces of certain 
other proteins but by no means with any face of 
any protein. Thus specificity in protein surfaces 
leads naturally to specificity in interlinks and in- 
terlinking will in general be by means of R-groups 
located on this or that face of each unit. On this 
basis, protein units may be expected to form pro- 
tein particles (as indeed Svedberg has already 
found in many cases) and structure systems of all 
degrees of complexity as T. Brailsford Robertson 
(in 1918) long ago' suggested. Such structure 
systems in the present picture will consist of rigid 
skeletons held together by interlinking groups. 
One main feature" of all the biologically-function- 
ing systems we are considering, namely their flex- 
ibUity, is now at once interpretable. A molecule 
such' as methane consists of atoms held in definite 
directions and at definite distances, allowing very 
little flexibility within the molecule. But an as- 
sociation of protein units into a protein particle or 
framework is of a totally different type so far as 
its mechanical properties are concerned. .\ny in- 
terlinking of protein units by means of R-groups 
allows some flexibility of the resulting structure. 
This result offers a reasonable and straight-for- 
ward interpretation of the deep-lying dichotomy 



found in cytoplasmic structure. On the one hand, 
this picture of the nature of association of pro- 
teins shows that it will be extremely specific, in 
so far as it involves the compatibility of patterns 
of R-groups. Association, being dependent upon 
R-group constellations fitting into each other suffi- 
ciently well to form a stable combination, is neces- 
saril)' conditional upon the maintenance intact of 
the characteristic skeleton, i.e., upon the condition 
that the native protein structure is preserved. Yet, 
on the other hand, such associations of protein 
units will give systems with definite flexibility. 
And this flexibility, let it be noted, is conjoined 
with and not in any sense incompatible with ex- 
treme specificity which is inherent not only in the 
individual units and their disengaged faces, if any, 
but also in the pattern of the various interlink 
types in the system as a whole. Putting these two 
superficially incompatible characteristics of speci- 
ficity plus rigidity of the skeletons and flexibility 
of the R-groups together we have a unit which 
promises well as the chief "structure-forming" in- 
gredient in biological structure systems. 

Geometry IV ill See Us Through 

From a totally different field of work, my own 
field of mathematics, there issues the same picture. 
This synthetic-geometrical attack has excited so 
much controversy from such able critics, con and 
pro, that there seems little doubt that, even if 
some of the arguments in favor of the theory may 
have so far been missed, practically all the ob- 
jections which can be l^rought against it have been 
meticuously tabled, a great advantage for any who 
might wish impartially to assess the situation re- 
garding native protein structure. On examination 
( see Langmuir, Proe. Ph\s. Soc. London, 51. 592, 
1939: Wrinch, J .A.C.S..'63, 330, 1941) these ob- 
jections seem to be lacking in force or cogency. 
Neither do they gain in these respects from con- 
stant repetition by those who thought them up, 
or by interested bystanders or workers in totally 
diff'erent fields, who didn't. Perhaps the most 
tenuous of the attempted onslaughts is that hav- 
ing to do with bond energies (known to be vari- 
able in different atomic environments) which 
might be possessed by a structure which has never 
been synthesized (so that no thermal measure- 
ments on it can be made) and their relation to a 
few thermal measurements on a single protein 
(denatured trypsin) whose structure is totally un- 
known. But the point which concerns us particu- 
larly in this new field is that the objections have 
focused attention upon details of the particular 
"fine structure" in terms of which the general idea 
of closed fabric structures for the globular pro- 
teins was clothed for expository, exploratory and 
general mathematical purposes. The actual fine 
structure or atomic pattern is, of course, quite in- 



August 9, 1941 



THE COLLECTING NET 



127 



susceptible of proof or disproof by gross chemical 
techniques in view of the lability of the native 
protein. X-ray analysis at first sight appeared to 
offer some hope of clinching the matter, and in- 
deed it was shown by Langmuir and myself that 
the predicted structure for insulin is compatible 
with the only X-ray data available. However in 
view of the subsequent pronouncement by a dis- 
tinguished X-ray authority that these data are too 
poor to be used for this purpose, this aspect of 
the matter is held up for the moment until data 
which these experts consider of value shall be 
available to test each and all of the predicted 
structures. 

I have stated that this concentration of the 
interest of my distinguished critics and others 
on the "fine structure" aspect of the work 
is of special interest to us here and now, for the 
following reason. The essential point of the work 
has, apparently, been entirely missed and it is this 
which is relevant to the protein structure systems 
which we now have under consideration, namely 
the possibility that the essence of these large 
structural units of proteins lies in a characteristic 
atomic fabric bent around to form a closed sur- 
face, a suggestion which could have a powerful in- 
fluence in the design of experiments. The emer- 
gence from the fields of physical chemistry, pro- 
tein crystallography and immunology of the gen- 
uine megamolecule, a structure consisting of thou- 
sands or millions of atoms which yet possesses 
definite physical, chemical and above all specific 
biological properties poses a new problem to 
physics and chemistry. The crisis is resolved and 
a beautiful new field of work is opened when it is 
seen that any patterned fabric of atoms gives the 
clue to this phenomenon. For any fabric of atoms 
will by its own pattern determine the types of 
closed surface which the metrical requirements of 
its constituent atoms will allow it to build. If the 
pattern of the fabric is a small pattern, then the 
resulting closed structures will give molecules of 
low molecular weight, like hexamethylene tetra- 
mine. But if the building units essentially require 
a large repeat in the pattern, as is the case with 
amino-acid and imino-acid residues, then the mo- 
lecular weights will be large. But, whether the 
molecules are large or small, the number of atoms 
or units in the skeletons will be determinate and 
a reason is then seen why even enormous protein 
structures should consist of perfectly definite num- 
bers of residue units. Indeed we can see one step 
further and predict that appropriate residue num- 
■ bers for protein skeletons should fall into definite 
series, necessarily quadratic functions of the na- 
tural numbers, since we are concerned solely with 
surfaces, i.e., two-dimensional distributions. It is 
on this basis that atomic fabrics which have been 
suggested in my investigations have yielded 72 as 
the first skeleton residue number, 288 as the sec- 



ond and so on, as the residue numbers for native 
protein units and multiples of any of these for 
protein particles, numbers which accord well with 
the molecular weights found by Svedberg. 

There are other striking consequences of this 
very simple idea. In a sense there is, as our col- 
league in China, Dr. Wu, was perhaps the first to 
emphasize, some general structural character 
which may be regarded as the essence of the pro- 
tein character. Yet there are astronomically 
many diff'erent proteins, all made up of a strictly 
limited variety of building stones. The cage or 
perhaps some other closed fabric structure like an 
anchor ring, with its obvious significance in terms 
of denaturation, according to which any break- 
down of the closed structure destroys the native 
character, may be, I submit, the essential protein 
character. So long and only so long as the R- 
groups have their roots held in this spatial pat- 
tern, is the specificity of the molecule retained. 
The ways in which the fixed number of places for 
residues in this highly organized, multiply-con- 
nected atomic skeleton can be filled by different 
complements of the residue varieties, or even by 
the same complement, are, very evidently, ex- 
tremely large in number. Each and every pattern 
of arrangement connotes potentially one very 
highly individualized and specific native protein 
unit. It is plain on this theory how the saine skel- 
eton can be used for the construction of native 
proteins with very diverse physical or chemical 
properties. Thus a skeleton having some definite 
number of residues may give rise to a molecular 
weight class — necessarily of wide spread, if any- 
thing like the available varieties of individual resi- 
due weights is made use of — containing proteins 
entirely distinct in their biological functions and 
chemical and physical identities. This situation is 
in fact realized in Svedberg's molecular weight 
class at about 36,000 for which the residue num- 
ber of 288 was predicted in 1936 on the basis of 
the cyclol theory. This class contains lactoglobu- 
lin, pepsin, insulin, chymotrypsin and a number 
of other entirely distinct protein species. 

A further aspect of the work requires a word 
of comment, in view of the necessity of relating 
any proposed structures to known facts regarding 
the high capacities and low conductivities of some 
biological structures. I refer to the fact that the 
fabric cages, which are here suggested, break 
down into monolayers with the utmost ease, (in- 
dicating once again the dependence of their sta- 
bility on interlinks with other molecules, water in 
this case) and to the possibility that the prodigi- 
ous insolubility of these monolayers may indicate 
that the native protein had in its interior a con- 
siderable complement of hydrophobic R-groups, 
shielded by the skeleton from contacts with a 
watery world. 

(Concluded in Next Issue) 



128 



THE COLLECTING NET 



[ Vol. XVI, No. 144 



The Collecting Net 

A weekly publication devoted to the scientific work 
at marine biological laboratories. 

Edited by Ware Cattell with the assistance of 
Boris I. Gorokhoff and Judy Woodring. 

Entered as second-class matter, July 11, 1935, at 
the U. S. Post Office at Woods Hole, Massachusetts, 
under the Act of March 3, 1879, and re-entered, 
July 23, 1938. 

NOMINATIONS FOR TRUSTEES 

The Annual Meeting of the Corporation of the 
Marine Biological Laboratory will be held in the 
atiditoriuni of the Lal:)oratory on Tuesday, Au- 
gust 12, at 11 :30 A.M., for the election of Officers 
and Trustees and the transaction of other busi- 
ness. The Trustees will convene the same morn- 
ing before the Corporation meeting and again in 
the afternoon. 

The Nominating Committee of the Corporation 
of the Marine Biological Laboratory has posted 
the following slate : 

For Trustee Emeritus — W. J. V. Osterhout 
For Treasurer of the Cor/^oration — Lawrason 

Riggs, Jr. 
For Clerk of the Corporation — Philip B. Arm- 
strong 
For Trustees of the Class of 1945— W. R. Am- 

berson, S. C. Brooks, W. C. Curtis, H. B. 

Goodrich, I. F. Lewis, R. S. Lillie, A. C. 

Redfield, C. C. Speidel 
For Trustee of the Class of 1943 to replace W. J. 

1'. Osterhout— G. H. A. Clowes 
For Aleinbership in Executive Committee — D. E. 

S. Brown, B. H. Willier 
Drs. Clowes and Brooks are proposed for 
Trusteeship for the first time; the other seven 
men are presented for reelection. 

ADDITIONAL INVESTIGATORS 

Addison, W. H. F. prof, normal histol. & emb. Penn- 
sylvania Med. 

Auger. C. Rockefeller Inst. Br 206. 

Bongiovanni, A. M. grad. biol. Villanova. Rock 3. 

Bowser. E. R., Jr. asst. fel. biol. Pittsburgh. Rock 7. 

Bowen. E. prof. biol. Gettysburg (Pa.). 

Deyrup, Ingrith grad. zool. Columbia. CM Phys. 

Green, J. W. grad. phys. Pennsylvania. Br 205. 

Gudernatsch, F. prof. biol. New York. Br 344. 

Kaylor, C. T. instr. anat. Syracuse Med. Br 226. 

Magnus-Levy, A. res. assoc. Yale. lib. 

Loewi. O. res. prof. Pharmacol. New York Med. lib. 

Maurer. Jane techn. Yale Med. Br 109. 

Meed, Margaret R. grad. biol. Brown. Br 312. 

Memhard, A. R. (Riverside, Conn.). Br 334. 

Nash, J. F. grad. asst. New York Med. Br 234. 

Oppenheimer, Jane M. instr. biol. Bryn Mawr. Br 
118. 

Ormsbee, R. A. grad. asst. Brown. Br 331. Ka 21. 

Packard, C. asst. prof. zool. Columbia. Br 102. 

Perkins, Patricia J. grad. asst. chem. Cincinnati. OM 
Phvs. WF. 

Rollason, H. D., Jr. grad. asst. biol. Williams. OM 
Phys. Dr 7. 

Sevag, M. G. asst. prof, biochem. Pennsylvania Med. 



DATES OF LEAVING OF INVESTIGATORS 

Costello, D. P Aug. 6 Mitchell, P. H Aug. 2 

Forbes, J Aug. 2 Mullins, C. P July 29 

Hager, R. P July 31 Runk, B. F. D July 26 

Henry, R Aug. 2 Schaffel, M July 29 

Hunninen, A. V Aug. 9 Spratt, N. T., Jr. July 29 

Levin, E July 29 Warner, E. N July 25 

REPRESENTATION BY INSTITUTIONS AT 
THE M. B. L. 

The following institutions are represented by 
three or more investigators registered (filled out 
registration blanks before August 8) at the Ma- 
rine Biological Laboratory this summer : 

Institution 1941 1940 

Pennsylvania 30 34 

New York 23 21 

Columbia 18 22 

Yale 13 8 

Hopkins 12 8 

Harvard 9 7 

Ohio State 9 8 

Chicago 8 13 

Rockefeller Institute 8 9 

Michigan 7 4 

Princeton 7 5 

Brown 6 5 

Cornell 6 6 

Pittsburgh 6 7 

Cincinnati 5 4 

Lilly Laboratories 5 4 

Milton Academy 5 4 

Vassar 5 2 

Villanova 5 2 

Washington 5 5 

California 4 5 

California Tech 4 2 

Iowa 4 4 

Mt. Holyoke 4 2 

Queens 4 4 

Rochester 4 2 

C. C. N. Y 3 3 

Illinois 3 2 

Miami 3 2 

Minnesota 3 2 

Missouri 3 6 

Oberlin 3 3 

Syracuse 3 5 

Temple 3 3 

Toronto 3 5 

Union 3 4 

Williams 3 3 



CURRENTS 


IN THE HOLE 


At the following 
Time) the current 
from Buzzards Bay 


hours (Daylight Saving 
n the Hole turns to run 
to Vineyard Sound: 


Date 


A.M. 


P.M. 


August 10 ., 
August 11 ... 


6:34 

7:18 


6:54 
7:40 
8:27 


August 12 .... 


8:01 


August 13 .. 


8:47 


9:15 


August 14 .... 
Ausfust 15 .... 


9:35 

10:26 


10:08 
11:02 
11:58 
12:13 


August 16 .. 
August 17 .. 


11:19 


In each case the current changes approxi- 
mately six hours later and runs from the 
Sound to the Bay. 



August 9, 1941 ] 



THE COLLECTING NET 



129 



ITEMS OF INTEREST 



Dr. W. E. Martin, who is instructing^ in the 
invertebrate course, has been promoted from as- 
sistant to associate professor of zoology at De- 
Pauw University. 

Dr. Edgar J. Boell, who worked here last 
summer, has been promoted from assistant to as- 
sociate professor of biology at Yale University. 

Dr. Frank H. Johnson, instructor of biology 
at Princeton University, has been promoted to as- 
sistant professor. 

Dr. John O. Hutchens, who has been a grad- 
uate student in zoology at the Johns Hopkins 
University, has been appointed instructor in phys- 
iology at the University of Chicago. 

A Research Institute of Endocrinology has 
been established at McGill University. Dr. J. B. 
Collip, who has been chairman of the department 
of biochemistry, will relinquish his position to 
become director of this Institute. 

On August 1, there were 278 investigators 
present at the Marine Biological Laboratory, as 
compared with 293 at the same time last year. 

The Trustees of the Woods Hole Oceanogra- 
phic Institution will hold their annual meeting on 
Thursday, the fourteenth of August. 

The second edition of Dr. James Mavor's text- 
book, "General Biology," was published in June. 
Dr. Mavor is professor of biology at Union Col- 
lege and is now working at the RIarine Biological 
Laboratory. 

At the staff meeting of the Oceanographic In- 
stitution on Thursday evening, Dr. Alfred C. 
Redfield spoke on "'The Distribution of Nutrient 
Substances in the Ocean." 

An informal statistical seminar will meet twice 
weekly during August in the Smoking Room at 
the Fisheries Residence with Dr. C. I. Bliss in 
charge. The first meeting was held on Thursday. 
This summer the design of experiments will be 
emphasized, but the program can be adjusted to 
fit the needs of those attending. 

M.B.L. CLUB NOTES 

In the ping-pong tournament, competition is 
planned in men's singles, women's singles, and 
mixed doubles, and perhaps in men's doubles and 
women's doubles. Persons wishing to compete 
should sign up on the chart at the Clul? House 
or see Teru Hayashi. 

The following composition will be presented at 
the phonograph record concert at the M.B.L. 
Club on Monday evening: Beethoven, "Missa 
Solemnis" in D (Opus 123), performed by the 
Boston Symphony Orchestra, conducted by Dr. 
S. Koussevitsky, with the Harvard Glee Club and 
the Radcliffe Choral Society. 



Professor Otto Loewi, who shared the Nobel 
Prize in 1936, arrived in Woods Plole last week 
with Mrs. Loewi. They will spend the balance 
of the summer here. 

Dr. James E. Kindred, professor of anatomy 
at the University of Virginia, who has worked 
here for a number of years, is supervising the 
building of a home in Charlottesville, Virginia, 
this summer. 

Dr. David Hand, assistant professor of bio- 
chemistry at Cornell University recently visited 
Woods Hole with Mrs. Hand on their 32-foot 
auxiliary cruiser Camara. 

Miss Helen S. Morris, teacher of biology at 
Evander Childs High School, in New York City, 
visited Woods Hole recently. She took the course 
in botany in 1927 and returned for research work 
for several summers afterwards. 

Mrs. Mildred W. S. Schram, executive secre- 
tary of the International Cancer Foundation, who 
worked at the Marine Biological Laboratory last 
year, is spending this summer at the Mt. Desert 
Biological Laboratory at Salsbury Cove, Maine. 

Dr. M. Perrot is working this summer at the 
Biological Laboratory at Cold Spring Harbor. 
During the academic year, he teaches at the Uni- 
versity of Missouri. 

Dr. Linville A. Baker was called to Anna- 
polis at the end of last week to take an ability 
test for second lieutenant in the Medical Admin- 
istrative Corps Reserve. He was away three 
days. 

Miss Grace Workman, who was here last 
year, was recently married to Dr. John W. Scott, 
of the Toronto General Hospital stafif. Dr. Scott 
took the physiology course at the Laboratory in 
1939. 

The Atlantis sailed on Wednesday on one of 
its regular cruises to the Gulf Stream under the 
direction of Dr. Alfred Woodcock. She is ex- 
pected to return in four weeks. 

M.B.L. TENNIS CLUB NOTES 

Play for the tournaments is now under way. 
Competition is going on in the men's and women's 
singles, and mixed doubles classes ; the finals in 
these classes are planned for Saturday, August 
16. The entries in the men's doubles are being 
held open for an additional week ; if eight teams 
sign up this class will also be included in the 
tournaments. 

The annual meeting will be hekl in the Old 
Lecture Hall next Wednesday at 7 :00. A pro- 
])osal to amend the by-laws to permit the mem- 
ijers of the immediate families of M.B.L. Corpor- 
ation members to hold office will be considered. 



130 



THE COLLECTING NET 



[ Vol. XVI, No. 144 



INVERTEBRATE CLASS NOTES 



With our second week at Woods Hole there 
has come a clearer understanding of the resources 
of the place, of how best to take advantage of 
them. In the lab one cannot examine all the 
material available ; with the aid of the lectures and 
alDundant mimeographed directions one can select 
material of particular interest. Our work on 
Coelenterates included a study of the transparent 
sea anemone NcinatostcUa. much of whose inter- 
nal structure is discernable without dissection. 
This ideal lab specimen with very limited geo- 
graphical distribution (Woods Hole, the Isle of 
Wight) was found in the Eel Pond three years 
ago by Dr. Bowen. The beautiful Ctenophore, 
Mnemeopsis, was also available. 

Our work with Platyhelminthes has revealed no 
beautiful specimens comparable to the comb-jelly, 
but Dr. Rankin has launched on a speedy efficient 
three-day defense of his pets, his accounts of 
weird life cycles being confirmed by our own lab 
.studies. 

Saturday's collecting trip to Lagoon Bridge on 
Martha's Vineyard provided an excellent antidote 
to a week spent in the lab. The somewhat dubi- 
ous weather made up its mind, by the time we got 
there, in our favor. How time flew as we scraped 
piles, passed mud and sand through the sieve, and 
turned over rocks ! The photographers among us 
were particularly busy during the picnic lunch on 
the beach. We got back to work before we had a 
chance to feel lazy, to be interrupted all too soon 
two hours later by the whistle announcing de- 
parture. So great was the force of the flowing 
tide that one of the boats was unable to point its 
bow into it. remaining hard against the bridge 
piles. By dint of much churning and a tow, we 
finally got headed for a sunny trip home. The 
trip was not complete, of course, until the speci- 
mens were placed in clean water, examined and 
labeled. 

BOOK REVIEW 



There were few people to be seen in the lab 
Monday evening. On this occasion, the hard hit- 
ting invertebrate team, paced by Bob Corder's 
home run in the first inning, trounced the highly 
touted crew team, ten to two. Much credit is due 
to the four-hit pitching of the Wabash star Howie 
Miner. \^'hile the crew was popping flies, the 
"Invert" Sluggers were busy collecting their six- 
teen hits. 

Conspicuous feature of the crew's game was the 
sparkling and vociferous play of catcher Jasper 
Trinkhaus. With the restoration of their original 
line-up, the crew expect to "clean the bases like 
(they) clean the lab." 

Not all the talk in the lab is of worms. There 
is proud papa Hauschka's delight in his 16-month 
old child's unmistakable flair for biology which 
has manifested itself in the latter's eating algae 
on the beach. It is now an acknowledged fact 
that Ann \\'eber was 1940's champion woman 
archer. They will never stop twitting Howie 
Miner about the letter he received from Gypsy 
Rose Lee. Arthur Culbertson injured his knee 
rushing to the dock to get a better glimpse of 
some interesting vertebrates on board an outgoing 
boat. Six members of the class, none of them ex- 
pert seamen, set out Sunday morning in one of 
Hilton's catboats. As they tacked back and forth 
they noticed squashes floating with the tide. It 
soon became apparent there was but one squash ; 
with all the maneuvering they scarcely kept up 
with the drifting vegetable. After a picnic lunch 
they commenced their return trip, riding the tide 
the other way. Just in sight of Woods Hole, one 
canny mariner discovered the wake was off the 
bo'cv! To make a long trip short, the suspecting 
Hilton arrived at 8 P. M. and towed them home. 

— Louise Gross and Bill Batchelor 



JOHANNSEN, 0. A. and F. H. BUTT. "Embryol- 
ogy of Insects and Myriapods". pp. xi + 462. 
New York, McGraw-Hill Book Company. 1941. 
$5.00. 

Periodically a work of real scholarship emerges 
from the welter of printed matter with which the 
scientific world is annually presented. That the 
present volume should fall under this classifica- 
tion is the more remarkable in that it is stated to 
be "the outgrowth of a course of lectures" ; most 
such outgrowths are either ijathological. being 
excised under the scalpel of the publisher's audi- 
tor, or insignificant warts which succumb to the 
caustic of public opinion. Johannsen and Butt are 
here presenting a source book on arthropod em- 
bryology which has reached, in its first printed 
edition, a degree of excellence which shows clearly 
that it has passed through many editions in manu- 



script. 

Two choices, either to write a clear account of 
the subject or to present both sides of all ques- 
tions, confront the author of a text of this nature; 
these choices form a dilemma on which has been 
caught many a promising volume. Johannsen and 
Butt avoid this impalement by attacking each 
horn separately. Their work is divided into two 
parts, the first ostensibly dealing with embryology 
under its biological subdivisions, and the second 
with the same subject under its taxonomic sub- 
divisions. The first part, however, gives a clear, 
well documented account of what appears to them 
to be the most probably true story. Doubtful 
points are cross referenced to the second part in 
which some 800 references have been reduced to 
a coherent and succint summary. Three hundred 
and seventy well-drawn and clearly labelled fig- 



August 9, 1941 ] 



THE COLLECTING NET 



131 



iires enrich the text: 

The reviewer, who is after all expected to criti- 
cise as well as praise, cannot refrain from one 
grumble. Chapter XII, which is called "Experi- 
mental Embryology", does not belong in a book 
of this general excellence. The authors appear to 
have been misled in their selection of material by 
considerations of what would appear most inter- 
esting to a vertebrate embryologist. Such purely 
entomological experiments as pupa grafting, trans- 



plantation of possible endocrine organs and the 
discovery of a pupal cuticle softening gland in 
certain coleoptera, have been ignored in favor of 
the few experiments which have been done on de- 
termination and organising centres. 

The publishers are to be congratulated on main- 
taining the high standard of production which has 
marked their previous publications in the zoologi- 
cal sciences. Peter Gray 



EFFECT OF AZIDE ON CYPRIDINA LUCIFERIN 

(Continued from Page 121) 



measured with an apparatus similar to that de- 
scribed by Anderson (/. Cell. Coinp. Physiol.. 
1933), involving a photoelectric cell, condenser, 
Lindemann electrometer, and potentiometer. 

There are a number of data now available 
which bear on the chemical structure of luciferin. 
First, it is definitely not protein and probably not 
lipoid, as indicated by its solubility characteristics 
(Anderson, /. Cell. Conip. Physiol.. 1936). An- 
derson has also called attention to the fact that its 
oxidation-reduction potential is similar to those 
of certain naturall)' occurring polyhydroxy ben- 
zene derivatives studied by Ball and Chen (/. 
Biol. Chem., 1933). The absorption spectrum of 
luciferin solutions (Chase and Giese, /. Cell. 
Comp. Physiol., 1940) bears a marked qualitative 
resemblance to those of certain naphtha- and an- 
thraquinone derivatives measured by Morton and 
Earlam (/. Cheiii. Soc., 1941). The visible ab- 
sorption spectrum of luciferin solutions undergoes 
changes during oxidation which may also indicate 
a quinoid structure (Chase, /. Cell. Comp. Physi- 
ol., 1940). Finally, Giese and Chase (/. Cell. 
Comp. Physiol.. 1940) found that cyanide com- 
bines irreversibly with purified luciferin to give a 
compound incapable of luminescence when lucifer- 
ase is subsequently added. They interpreted these 
results as indicating an aldehyde or keto group 
in the luciferin molecule as the point at which 
cyanide, and probably luciferase, acts. 

Chakravorty and Ballentine (/. Amcr. Chcni. 
Soc, 1941) have summarized most of the chemi- 
cal data on luciferin and, on the basis of these 
and of certain additional experiments of their own, 
have suggested a partial structure for the luciferin 
molecule, consisting of a hydroquinone nucleus 
and a keto-hydroxy side chain. The keto group 
in the side chain would be the site of action of 
cyanide and of luciferase, the hydroxy group 
would be the point of action of benzoyl chloride 
and other similar reagents, and the hydroquinone 
nucleus would be capable of reversible oxidation 
and would thus represent this characteristic of 
luciferin. 

Giese and Fisher (unpublished observations) 
have found that sodium azide markedly inhibits 
the luminescence of luminous bacteria and this 
observation prompted the present experiments on 



effects of azide on the luminescent reaction be- 
tween Cypridina luciferin and luciferase. At pH 
6.6 sodium azide, in concentrations from 0.01 to 
0.1 M., inhibits the luminescent reaction reversib- 
ly and in such a way that a combination between 
the azide and the luciferin is suggested. At pH 
5.4 a smaller concentration of sodium azide pro- 
duces the same degree of inhibition of lumines- 
cence. Comparison of results at these two pH's 
indicates that it is the undissociated HN3 which 
is effective, rather than the NaNa. When the 
data are tested in terms of the mass law equation 
by plotting logarithm of percent total luminescence 
obtained minus logarithm of percent total lumin- 
escence inhibited against logarithm of sodium 
azide concentration, they are found to be fitted by 
straight lines with slopes approximately equal to 
unity. This might indicate that a single azide 
molecule or ion combines with a luciferin mole- 
cule to produce the observed effect. 

The following interpretation of these results in 
terms of the supposed structure of the luciferin 
molecule is intended as purely tentative. Schmidt 
(Chcin. Abstr., vol. 19, p. 3248), Franklin (/. 
Amer. Chem. Soc., 1934), and others have de- 
scribed the effect of hydrazoic acid on certain al- 
cohols, aldehydes and ketones, for example, but 
the conditions of the reaction are very drastic 
compared with those of the present case and it 
seems unlikely that such a reaction would occur, 
even though the experiments with cyanide have 
indicated that an aldehyde or keto group may be 
present in the luciferin molecule. Fieser and 
Hartwell (/. Amer. Chem. Soc, 1935) have re- 
ported the action of hydrazoic acid on benzo- and 
naphthaquinones. An azido-hydroquinone is first 
formed and this may change to an amino-quinone 
with liberation of nitrogen. This type of reaction 
does not seem to require very drastic methods 
and, in view of the oxidation-reduction potential 
of the luciferin system and the absorption spec- 
trum of purified luciferin solutions, it seems pos- 
sible tentatively to interpret the effect of azide on 
luciferin as indicating a reaction with a quinoid 
structure. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on 
July 29.) 



132 THE COLLECTING NET [ Vol. XVI, No. 144 



To be ready for fall use — 

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THE COLLECTING NET 



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134 



THE COLLECTING NET 



[ Vol. XVI, No. 144 



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THE COLLECTING NET 



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136 



THE COLLECTING NET 



[ Vol. XVI, No. 144 




Haifa Glass Bead Says, "She'll Live" 



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INTERPRETATIONS OF EFFECTS OF CO 
AND CN ON OXIDATIONS IN CELLS 

Dr. Matilda M. Brooks 
Research Associate, University of California 

I. Effects on Rate of Oxygen Coiismnption. 

In these preliminary experiments, eight stages 
in the development of sea urchin or starfish eggs 
were used to see what effects certain accelerators 
or inhibitors of O2 consump- 
tion would have. The stages 
were unfertilized eggs, first 
cleavages, 32-cell to morula, 
blastula, early gastrula, late 
gastrula. early pluteus, and 
late pluteus in the case of Ar- 
bacia, and unfertilized eggs 
and first cleavages in the case 
of Asterias. Methylene blue 
which is assumed to be an ac- 
celerator and KCN and CO 
which are assumed to be in- 
hibitors of respiration were 
used. The experiments were 
done in triplicate by the War- 
burg method using aliquot por- 
tions of a suspension of eggs 
or larvae in sea water for each 
of the 12 Warburg vessels 
operated simultaneously. Con- 
centration of methylene blue 
was .00012 M ; of KCN, .00025 M and of CO 
as near to 100% as was possible to obtain. 

It was found that (Continued on page 150) 



M. %' If. €alcll^a^- 

MONDAY, August 18, 8:00 P. M. 

Lecture: Dr. R. W. Wilhelmi: "Ser- 
ology Applied to Pi'oblems In- 
volving Phylogenetic Relation- 
ships and Evolution of Animals." 

TUESDAY, August 19, 8:00 P. M. 

Seminar: Dr. D. Nachmansohn: 
"Electrical Potential and Activity 
of Choline Esterase in Nerves"; 
Dr. A. Claude: "Chemical Compo- 
sition of Mitochondria and Secre- 
tory Granules"; Dr. D. Wrinch: 
"Native Proteins and the Struc- 
ture of Cytoplasm." 

THURSDAY, August 21, 8:00 P.M. 
Lecture: Dr. H. Dam: "The Biolog- 
ical Significance of Vitamin K." 

FRIDAY, August 22, 8:00 P. M. 

Lecture: Dr. O. Meyerhof : "Nature, 

Function and Distribution of Phos- 

phogens in the Animal Kingdom." 



THE ORIGIN OF GAPS BETWEEN 
SPECIES 

Dr. Ernst Mayr 

American Museum of A^atural Historw 
Neiv York, N. v. 

Some day, when the history of the science of 
evolution will be written up, the historian will 
have to distinguish various periods. The fact that 
we seem to be right at the be- 
ginning of a new one of such 
periods makes the work in the 
tield of evolution so particu- 
larly interesting. Up to Dar- 
win, evolution was, more or 
less, a speculative subject. The 
publication of "The Origin of 
Species" in 1859 started a sec- 
ond period in the history of 
the science of evolution. In 
this period all the heavy artil- 
lery of the biological disciplines 
of paleontology, taxonomy, 
com]3arative anatomy and em- 
bryology was assembled to de- 
molish any doubts and objec- 
tions that were raised against 
the principle of evolution. 
Final victory of this battle was 
gained by the end of the cen- 
tury and the discussion moved 
from the question : Is there evolution ? to the 
question : How does evolution proceed ? This is 
the question on which the student of evolution 



TABLE OF CONTENTS 



The Origin of Gaps Between Species, Dr. 
Ernst Mayr 137 

Interpretations of Effects of CO and CN on 
Oxidations in Cells, Dr. M. M. Brooks 137 

Aging Phenomena and Factors Influencing 
Longevity in Mactra Egg Cells, Dr. Victor 
Schechter 143 



Proteins in Action (Cent.), Dr. Dorothy 

Wrinch 143 

Biology at the University of Texas 146 

Additional Investigators 146 

Drtes of Leaving of Investigators 146 

Items of Interest 147 

Invertebrate Class Notes 148 




THE MARINE BIOLOGICAL LABORATORY 



August 16, 1941 ] 



THE COLLECTING NET 



139 



has concentrated since 1900. 

Darwin entitled his epoch making- work not the 
"Principles of evolution" or the "Origin and de- 
velopment of organisms." No, he clearly realized 
that the species problem was the core of the proli- 
lem of evohition, and he, therefore, called his book 
boldly "On the Origin of Species." This is a fact 
that should not be forgotten. 

Speciation is the name we give to an evolution- 
ary process by which one former species is broken 
up into several separate units which have changed 
sufficiently to be considered different species. 
There are thus actually two processes involved : 
the development of diversity as well as the estab- 
lishment of discontinuities between the newly 
evolving species. 

The development of diversity is the more ob- 
vious one of these two aspects of evolution and the 
work of the geneticists between 1900 and 1930 
was primarily devoted to an elucidation of this 
aspect. This work consisted in an analysis of the 
differences between various species and other na- 
tural units, and in an investigation of the genetic 
basis of these differences and of the origin of new- 
variants, the so-called mutations. Some phases of 
this work are still incomplete, in particular what 
we might call the physiology of mutation and gene 
action, but the major principles of chromosomal 
inheritance and mutation are now well established 
and generally accepted. 

However, those geneticists who were most in- 
terested in the evolutionary aspects of genetics, 
men like Dobzhansky and Sewall Wright, were 
the first to realize that even if we knew everything 
about variation and mutation, we would still be 
far from an understanding of the origin of species. 
After all, it is quite thinkable that such variation 
would only lead to a single interbreeding com- 
munity of immensely variable individuals. But 
this is not what we find in nature. 

The taxonomist, who approaches the diversity 
of organic life, finds that it is not one homogen- 
eous mass of genetically different individuals, but 
rather that it can be broken up into smaller, na- 
tural units, the so-called taxonomic categories of 
families, genera, species, etc. Each one of these 
is more or less separated from the other cate- 
gories ; they form a series of discontinuities. Evo- 
lution is a continuous process, but the units pro- 
duced by evolution are discontinuous. 

I have already said that we seem to be at the 
beginning of a new phase of the study of evolu- 
tion and what I meant by this phase is the study 
of the origin of discontinuities. This is a typical 
border line field which must be studied jointly by 



the naturalist and ecologist, by the geneticist and 
taxonomist. 

Now let us study the question of interspecific 
discontinuities as exemplified in some of the fami- 
liar New England birds. The ornithologist unites 
for example, the thrushes in the genus Hylocichla. 
But if we examine the variation within this genus 
in more detail we find that it clusters very closely 
around five means to which we apply the familiar 
names wood thrush, veery, hermit thrush, gray- 
cheeked thrush and olive backed thrush. They 
are all quite similar to each other, but two or 
three of them may nest in the same wood without 
the slightest intergradation. Not a single hybrid 
is known between these five common species. 

To take another example : if we sail out to the 
tern islands of the New England coast, we will 
find the three species : the common tern, the 
roseate tern, and the Arctic tern. All three 
species again are very similar, so similar indeed 
that only an expert can tell them apart in the 
field, but there is not a single intergrade or hy- 
lirid known between the three species. Moreover, 
they differ in their ecology and behavior patterns. 
In all these cases the related species are separated 
by clearcut discontinuities or what Goldschmidt 
would call "bridgeless gaps." 

Such observations lead us to the assumption 
that the species is a basic and objective natural 
unit. The majority of the animal taxonomists, 
particularly of the better known groups, subscribe 
to this opinion. But there is a considerable min- 
orit}' of authors who say just exactly the opposite. 
To them, the individual is the only unit in nature 
which possesses any reality while species are 
merely abstractions. They claim that all organ- 
isms form a continuity, which the taxonomist 
chops up into species merely for the sake of con- 
venience, to be able to handle them better in the 
museum drawers. 

The real test of the question, whether species have 
an objective reality or not. can only be made in 
groups which are taxonomically well known. 
Birds are, unquestionably, such a group. Now if 
we ask ourselves, are species objective units in 
birds or not. we must ask at once, what are the 
criteria by which the objectivity of a systematic 
unit can be determined? Thinking this over, we 
come to the conclusion that such a unit is objec- 
tive or real, if it is delimited against other units 
by fixed borders, by definite gaps. 

Now we mentioned already that such definite 
gaps exist, if we compare all the New England 
thrushes or all the New England terns with each 
other. There is no question that they are good 



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140 



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[ Vol. XVL No. 145 



species. The same is true, with few exceptions, 
whenever we comj^are species tliat occur at the 
same locaHties. what I call sympatric species. But 
there is a second type of relationship between 
species, namely between species that represent 
each other geographically, the so-called allopatric 
species. Let us look at a bird with which most 
of you are familiar, the junco, the little gray spar- 
row, with the white outer tail-feathers, that comes 
to our feeding stations during the cold season. 
This bird occurs in a number of populations in the 
Alleghanies from Georgia north and in the con- 
iferous zone from New England to the Rock\' 
Mountains and to the northern tree limit. All 
these populations merge into each other and even 
though the junco of the southern Alleghanies is 
slightly different from the Canadian bird, there is 
no doubt that it belongs to the same species. The 
trouble starts in the Ftocky Mountains. Here we 
lind a series of mountain forms which extend 
from .Alaska south as far as Panama, and some 
of which are completely isolated from all other 
junco po]3ulations. Most of them, however, inter- 
breed where they meet each other, even though 
some of these forms are so distinct that the taxon- 
omist prefers to call them different species. Clear- 
ly these allopatric species are not separated from 
each other by bridgeless gaps ; they can not be 
delimited in an ol^jective way. Such a situation 
is. of course, not really surprising because it 
would be exceedingly difficult to understand evo- 
lution if we did not have such cases of incom- 
liletely separated species or incipient species. We 
have learned from this discussion that species can 
lie delimited from two kinds of other species, from 
svm])atric species, where the gap is complete, and 
from allojiatric six'cies. where the gaj) is frequent- 
ly incomplete. 

Let us now look a little more closely at this 
delimitation and begin with the simple situation 
of the gap between sympatric S])ecies. In well 
worked groups, such as birds, the taxonomist does 
not encounter any serious difficulties when he tries 
to delimit sympatric species against each other. If 
there are any difficulties the final analysis usually 
reveals that either ( 1 ) several stages or phases of 
a species are so different that they had been mis- 
taken for different species or (2) just the oppo- 
site, that several species that occur in the same 
locality are so similar that they were considered 
stages of one species. 

Illustrations for the first kind of difficulty are 
easy to find. They are, however, not of great 
theoretical importance. More interesting is the 
other class of difficulties where pairs or even 
larger groups of related species are so similar 
that they are generally considered as belonging to 
one species, until the final analysis clears up this 
mistake. A number of such recently analyzed 
species groups are known in the genus Droso- 



phila, such as for example Drosophila "ohscura" 
and D. "affiiiis." The species of the flycatcher 
genus Euipidonax are the closest parallel to this 
that we can find among North American birds, al- 
though the analysis of this case was completed 
more than a generation ago. 

"Sibling species," as I call these morphological- 
ly similar species, are common in some classes and 
orders of animals and rare in others. Their im- 
])ortance is the following : In the bygone days of 
a purely morphological species definition they 
were considered races of one species. And since 
many of these species were originally discovered 
on the basis of biological or ecological differences, 
they were originally established as biological races. 
The literature is full of references to "biological 
races" of this kind, but closer analysis showed in 
a great majority of the cases that these so-called 
"biological races" are perfectly good species on 
the basis of every single criterion except the mor- 
]ihological one. The sibling species around the 
Malaria mosquito. Anopheles maciilipennis. are 
])erha]5s the most instructive illustration of this 
]irinci])le. .Ml the various races described by 
earlier workers, such as atro parvus, mcsseae, 
iiiclaiiooii. sacharoz'i differ not only in their ecol- 
ogy and habits, but also fail to interbreed in na- 
ture, and are completely or partially sterile if they 
are artificially crossed in the laboratory. A more 
painstaking analysis has recently revealed fairly 
constant morphological differences between some 
of these species. 

I have discussed these sibling species in a little 
more detail because their existence has been 
((noted as evidence in support of the idea of sym- 
]5atric speciation, a subject which we will pres- 
ently examine more closely. Such an assumption 
is, however, not justified. There is much reason 
to believe that even in sibling species speciation 
l^roceeds by geographical variation. 

Geographical variation 
Let us now examine the gap that exists between 
allopatric, that is, geographically representative 
s]iecies. This can best be demonstrated by ex- 
amining the distribution maps of some bird species 
such as Pachycephala and Zosterops. We can 
summarize these cases and the previously men- 
tioned case of junco, by stating that we have in 
all these cases geographically representative forms 
which, although morphologically well character- 
ized, are clearly descendents of one common an- 
cestor and which have not yet lost their sexual 
affinity. This is proven by the fact of unlimited 
hvbridization wherever such species ranges meet. 
Morphologically, such forms are species, biologic- 
ally they are not. These groups of allopatric spe- 
cie's are' the final results of a process which we call 
geographical variation. Let us now examine the 
beginnings of this process. 



August 16, 1941 ] 



THE COLLECTING NET 



141 



If a iuiml)er of populations of any widespread 
species are compared with each other, it is usually 
found that tliey show certain frenetic dififerences. 
This has lieen proven alumdantl}' by the work of 
Sumner and Dice on Pcroniyscus. of Goldschmidt 
on Lyiiianti'ia dispar. of Dobzhansky, Timofeeff, 
Patterson, and others on Drosophila, to mention 
just a few of many similar investigations. The 
taxonomist finds the same in his work. Some- 
times these dififerences are only slight differences 
of size, very often they are more pronounced. 

Populations, with definite geographical ranges 
and fixed systematic characters, are called geo- 
graphical races or subspecies. As long as these 
suljspecies are distributed continuously as in the 
case of the mainland races of Myiolcstcs, there is 
no question that they all belong to the same spe- 
cies ; they are one continuity. But wherever geo- 
graphical or ecological factors produce barriers, 
we find allopatric species gaps and we must use 
our judgment if we want to decide whether or not 
we consider them species. This is, for example, 
true for the insular races of Myiolcsfes. 

Such isolated populations are of the greatest 
importance for the question of evolution. When 
they first become isolated they may not be differ- 
ent at all from the parent population from which 
they split off. However, the three factors of 
mutation, difference of selective factors and differ- 
ences of random gene loss will produce an in- 
creasing difference between the two populations. 
After some time the difference will be sufficiently 
large to be noticed by the taxonomist. He says 
the i.solated population is a different subspecies. 
Eventually, this subspecies will be different 
enough to be called a different species by some 
taxonomists. But is it really a different species ? 
Are there any criteria by which we can determine 
this ? The old-fashioned taxonomist will say 
yes. there is a good criterion. If this isolated pop- 
ulation is characterized by a well defined charac- 
ter which in its variation shows no overlap with 
the ]>arent po])ulation, it is a species. But such a 
purely morphological species definition is not ac- 
ceptable to the modern biologist, l:)ecause species 
are, after all, not the creation of museum taxono- 
mists, but products of nature. 

We should, therefore, let nature make the deci- 
si<in what is to l^e considered a species, and not 
rely entirely on the subjective judgment of the 
taxonomist. Reproductive isolation or the lack 
of it, is nature's criterion. In other words, if two 
forms meet in nature and intergrade or freely in- 
terbreed, they must be considered as belonging to 
one species. If, on the other hand, two forms 
nuet in nature and act like two different species, 
that is they do not interlsreed, we must consider 
them as good species. A species definition based 
nil these Ijiological criteria would have to be for- 
mulated about as follows: "A species consists of 



(I f/roiip of populations which replace each other 
i/eo(jraphica!ly or ecologically and oj zdiich the 
neighboring ones intergrade or inlerhrecd tvher- 
erer they arc in contact, or zvhich arc potentially 
capable oj doing so in those cases where contact 
IS prevented by geographical or ecological bar- 
riers." 

On the other hand, we do not include in the 
same species isolated populations that have 
changed to the point that they are no longer cap- 
able of interbreeding with the parent race. We 
are not concerned here with the question what 
kind of differences must develop between these 
isolated populations, to bring about this repro- 
ductive isolation. This is a question to be ex- 
amined by the geneticist. The naturalist believes, 
and the majority of the geneticists endorse this 
idea, that the mere accumulation of genetic dift'er- 
ences will eventually lead to a difference suffici- 
ent to prevent interbreeding. 

What the taxonomist is interested in is the 
]3urely descriptive fact that the isolation of certain 
]iopulations together with geographical variation 
will in due time lead to the formation of new 
species, in fact that this is the normal process of 
species formation. This is the concept held by 
the vast majority of the thinking taxonomists, but 
this point of view has by no means remained un- 
challenged. The most severe critic has probably 
Iieen Professor Goldschmidt in his recent book 
"The Material Basis of Evolution." 

Goldschmidt argues that the small variants 
which ordinary gene mutations produce are in- 
sufficient to account for the basic differences 
which exist between species. He, therefore, pos- 
tulates that species must originate by revolution- 
ary changes in the genetic make-up, his so-called 
systemic mutations, and not by a gradual accumu- 
lation of differences with the help of geographical 
isolation. It would lead too far, in this connec- 
tion, to attempt a point by point refutation of 
Goldschmidt's argumentation. I will merely point 
out that Goldschmidt confuses completely the two 
kinds of gaps between species, namely between 
sympatric species and between allopatric species. 
The gaps between sympatric species are absolute 
or bridgeless as Goldschmidt says correctly, other- 
wise they would be no good species : the gaps be- 
tween allopatric species are relative, otherwise 
there would not be any evolution. 

Goldschmidt claims that geographical variation 
leads only to diversification within the species, but 
not to the formation of new species, as the taxon- 
omist claims. Instead of demolishing Gold- 
schmidt's arguments we shall try the more direct 
a])]3roach and marshal the evidence of the taxono- 
mist in favor of the importance of geographical 
speciation. These arguments are : 

( 1 ) Every character that is known to separate 
good species has also been found on close study to 



142 



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[ Vol. XVL No. 145 



vary geographically. This concerns morphologi- 
cal and physiological characters and includes in- 
terspecific sterility. Far distant geographical 
races of some species may be partly or completely 
sterile with each other, while neighboring races 
are, of course, always completely fertile. 

(2) There is no criterion by which it can be 
determined whether isolated forms, such as Pach- 
ycephala, Boiiibiis and Zosterops, are subspecies 
or species. Geographical variation coupled with 
isolation, completely blurs the borderHne. 

( 3 ) Well isolated forms may show extreme 
morphological specializations which under ordin- 
ary circumstances would be considered as of 
generic value. 

(4) Isolation on islands leads to species for- 
mation as proven by double colonizations. 

(5) Slight overlaps may occur after the 
breakdown of the isolating barriers. 

(6) If the final links of a chain of races over- 
lap, they may act like good species, even though 
connected by a series of intergrading populations. 

All these cases prove that if a population of a 
species becomes isolated and remains large enough 
in this isolation, it may acquire characters in this 
isolation which may enable it to e.xist as separate 
species beside the parental species, by the time the 
isolation barrier has broken down. The discon- 
tinuity which had originally been artifically main- 
tained by geographical barriers becomes physio- 
logical and reproductive and the one time single 
species has developed into two. 

Occasionally it happens that the geographical 
barrier breaks down before -the reproductive iso- 
lation has become completed. The result is ex- 
tensive hybridization in the zone of contact. A 
celebrated case is that of the flickers. 

Sytiipatric Spcciation 

1 have treated speciation, up to this point, en- 
tirely from the point of view of the ornithologist. 
Wt know that in birds all incipient species have 
to go through the stage of geographical races. 
New species can develop only in geographical iso- 
lation. We might call this "geographical" or "al- 
lopatric" speciation. The question remains 
whether it is not possible in other animal groups 
to have speciation through ecological specialisa- 
tion. The discontinuity, which would eventually 
lead to a gap between good species, would have 
to develop, in such cases, within the population 
of one geographical district. We might call this : 
sympatric speciation. 

Some authors believe that this type of specia- 
tion is very wide-spread. Careful recent studies 
have tended to disprove this assumption. Many 
cases that were formerly quoted as proof for sym- 
patric speciation have now been unmasked as 
sibling species. Some authors forgot that sym- 
patric speciation can operate only if some ecologi- 



cal device exists which makes the cross-mating of 
the two diverging lines impossible. Such isolating 
mechanisms are, for example, distinct and non- 
overlapping breeding seasons, or in the cases of 
parasitic or monophagous species strict host spe- 
cificit}' with the mating taking place on the host. 
Enough such cases have been described to make 
me believe that sympatric speciation is of com- 
mon occurrence in certain animal groups with 
very specific ecological requirements, but almost 
completely absent in all other animal groups. 

Our knowledge on the process of speciation — 
that is, the establishment of discontinuous natural 
units — is still very obscure in many animal 
groups, for example in fresh water and marine 
forms. Considerable difficulties are also caused 
by the so-called cosmopolitan groups. The Clado- 
cera, rotifers, tardigrades, to name just the most 
prominent ones, are groups that are composed 
primarily of cosmopolitan species. The same iden- 
tical species may be found on every single conti- 
nent and in every latitude from the Arctic to the 
Antarctic. This is not surprising if we remember 
that all these forms, or at least their eggs, can 
encyst. Such dry cysts can easily be carried by 
wind ciuTents clear around the world. The diffi- 
culty for the student of speciation lies in the fact 
that the continuous swamping of every population 
by new immigrants does not permit the establish- 
ment of new forms, at least so it seems. No ade- 
quate explanation for speciation in these families 
and orders has yet been advanced. There are sev- 
eral possibilities. One is that there is actually 
no longer any speciation in the most successful 
and most widespread of these species. Speciation 
is restricted to the more localized and, from the 
point of view of dispersal less successful species. 
The other possibility is quite diiiferent. Most of 
the above mentioned groups (the tardigrades are 
a notable exception) have parthenogenetic genera- 
tions sandwiched in between the bisexual ones. It 
is possible that a radically new mutation in a 
single individual affecting, for example, the mat- 
ing l)eha\-ior. can build up a sufficiently large 
population during the parthenogenetic period, to 
be able to survive afterwards as the ground stock 
of a new species. 

I have mentioned this case merely to indicate 
that there are many inisolved problems in the field 
of speciation. The geneticists have contributed 
their share, by unraveling a good many of the 
problems concerning the diversification which 
leads to new species. However, the origin of dis- 
continuities is a problem outside of this field, no 
matter how important it is in relation to genetics. 
And Dobzhansky says : "The aspect of discontin- 
uity should be emphasized, not because it is the 
more important of the two [processes of specia- 
tion], but because it is the less obvious one to 
the superficial observer." Actually many of the 



August 16, 1941 ] 



THE COLLECTING NET 



14.5 



laboratory workers do not seem to understand the 
significance of the fact that the vast variability of 
nature is not continuous, but broken up into a 
liierarchy of separate groups, of which at least the 
species has an objective reahty. 

The ornithologists, and to some extent also the 
students of mammals, butterflies and beetles, have 
gone a long way toward understanding how the 



discontinuities develop which precede the origin 
of new species in their groups. It will now be the 
task of the students of other animal groups to in- 
vestigate how far these findings arc applicable in 
their own field, or else what additional evolution- 
ary mechanism they can discover. 

(This article is based upon a lecture delivered at 
the Marine Biological Laboratory on July 31.) 



AGING PHENOMENA AND FACTORS INFLUENCING LONGEVITY IN MACTRA 

EGG CELLS 

Dr. Victor Schechter 
Assistant Professor of Biology, College of the City of Nezv York 



The unfertilized egg cells of the clam. Mactra 
solidissinia, go through an interesting series of 
morphological changes with age. The most out- 
standing of these are, in time series : 

( 1 ) Indentation of the cortical region proceeds 
until the egg is grotesquely collapsed. 

(2) Inability, upon insemination, to cast off 
polar bodies. 

(3) Loss of ability to cleave. 

(4) Lack of germinal vesicle maturation, upon 
insemination. Cortex still responds to sperm by 
partial rounding up. 

(5) Spontaneous germinal vesicle breakdown 
at extreme age. Polar bodies occasionally formed. 
Cortex rounds up. 

It is conceivable that these changes, on the 
whole, might be explained by a loss of osmotic 
materials within the egg, combined with a stiffen- 
ing of the cortex. The final spontaneous germi- 
nal vesicle breakdown could be due to a release of 
salts, like calcium, from the egg cortex to the in- 
terior. This hypothesis suggests the experimental 
procedure of varying the concentration of salts in 
the medium. 

The results are clear-cut, in showing : 

( 1 ) A prolongation of life in low calcium solu- 
tions. 

(2) A lesser prolongation of life in high cal- 
cium solutions. 

(3) The most marked prolongation in low cal- 
cium combined with high magnesium. 

The anomalous effect of high calcium is difficult 
to explain in terms of the working hypothesis 
adopted above. Its effect might be to decrease 
permeability and retard the flow of materials 
across the cell membrane. The effect of magne- 



sium can easily be thought of as due to the dis- 
placement of calcium in the egg cortex. It is also 
known thac the breakdown of germinal vesicles by 
radiation is inhibited by magnesium. 

There are other factors which are known to in- 
crease egg cell longevity. Among these are : 
acidity ; KCN ; low temperature ; thyroxin ; egg 
albumin ; alcohol ; glucose ; gum arable ; reduced 
bacterial numbers. 

That any of these are effective in any nutritive 
way seems ruled out by the facts that an activated 
egg cell will continue living without food at least 
through the embryonic stage ; and an unfertilized 
egg cell will live for a longer period when placed 
in sterile sea water. The relationship of egg cell 
longevity to the presence or absence of bacteria is 
an interesting problem in itself. With regard to 
the other factors listed above it may be significant 
that several of them affect viscosity and with vis- 
cosit3^ pH, iso-electric points and salt binding 
power are colligative properties. 

Although many interesting parallelisms may be 
drawn between grov\'th, differentiation, matura- 
tion and senescence in the short pre- and longer 
post-fertilization life of an egg it would be unwise 
to attempt to extend to the compound animal 
body, as a whole, the findings with regard to the 
relationship of calcium to aging in egg cells. This 
is obviously because of the role of calcium in many 
functional and organ-forming processes. As a 
general cellular phenomenon the hypothesis is 
worthy of further investigation. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on 

August 5.) 



PROTEINS IN ACTION 

Dr. Dorothy Wring h 

Professor at Amherst, Smith and Mount Holyoke Colleges 

(Continued from last issue) 



Bonds, Bridges and Links 

On the basis of this closed fabric structure for 
the native proteins, there seems then to be no dif- 
ficulty in interpreting and integrating the findings 
from the three important experimental fields we 



have discussed. To see rather clearly what the 
implications are regarding the ways in which such 
structural units can build structure systems, it is 
then only necessary to study what is known, from 
structure chemistry, as to how the R-groups, 



144 



THE COLLECTING NET 



[ Vol. XVL No. 145 



whose nature is well-known, may be expected to 
interlink or associate. There may, of course, be 
cases in which such interlinking is covalent in 
character, e.g., in the case of cystine residues or 
basic and acidic groups which have formed a pep- 
tide bond. There are, however, a considerable 
variety of other types which seem to be indicated. 
These include hydrogen bridges between gluta- 
mines and asparagines through their CO-XH;; 
groups, similar interlinks between acidic and basic 
groups, e.g., in glutamic or aspartic acid and argi- 
nine and, most important for our biological prob- 
lems, the bolting together of similarly ionized 
groups by means of divalent ions of opposite sign. 
It is well-known that hydrogen laridges have a 
definite energy, say 4 to 8 kcal./mole. Contrast- 
ing this with the energ>' in a carl)on-carbon bond, 
wliich we may take as (say) 60 kcal./mole. we 
see that such liridges are essentially weaker links 
than covalent links. Even the simultaneous for- 
mation of ( say ) 6 would give a total energy of 
less than a single C-C Ijond. The nature of the 
treatments by which such protein particles as 
horse hemoglobin, the seed globulins, the hemo- 
cvanins and the erythrocruorins can be dissociated 
into smaller units and particularly their mildness 
suggest that weaker links of this polar type, rather 
than covalent bonds, may be the means of asso- 
ciation in such cases. 

An inventory of all the types of intermolecular 
association discovered in crystal analyses calls our 
attention also to a second type, which is still 
weaker, namely that due to van der W'aals attrac- 
tions between hydrocarbon groups, such as e.xist 
in hydrocarbon crystals. Such association will 
de]iend u]ion a favorable ojjposition of what we 
may call the van der W'aals outlines of R-group 
Cdustellalions on op]50sed faces. While describing 
such a situation in terms of interlinks, it will be 
understood that the integrated effect cannot be 
analyzed into localized associations of individual 
atoms or links in the usual sense. 

In this and other aspects, it is interesting to 
notice the similarities and dissimilarities between 
these two categories of association to be expected 
when protein units associate by means of their 
R-groups. Both types are properly called "weak", 
if thev are considered in terms of individual pairs 
of R-groups and if our norm is the covalent bond, 
l)ut a sufficient amount of simultaneously opposed 
groupings permits an association of considerable 
stability. Next, in both cases the specificity of the 
]irotein surfaces is the all-important point. While 
some measure of association may occur between 
almost any protein surface and another, since 
acidic and basic groups are present in most cases 
and non-polar groups can also stick together, it 
will only be possible to have a comparatively 
stable association when the particular constella- 
tions of many R-groups are favorable. This is, 
to my mind, the essence of the protein as a "struc- 



ture-forming" component, to use Lillie's phrase. 
The s]iecific pattern of the protein itself deter- 
mines whether or not it will associate in stable 
combination with another specific protein or in- 
deed with any other molecule, such as a sugar, or 
a phosphatide. This fact is, I think, at the root 
of the non-antigenic character of certain molecules 
in certain cases. This also is the essence of the 
formation of antibody-antigen complexes. It ac- 
counts, too, for the way in which certain molecules 
and ions can change the permeability of cell mem- 
branes. The very low order of magnitude of the 
number of molecules required in such cases is of 
great interest. 

Specificity will also play a role in the van der 
Waals type of association. A highly organized 
structure like a .sterol may form a comparatively 
staljlc association with the surface of a protein if 
the R-group constellation has an appropriate mor- 
phology. The difference between the two types 
of association resides in the higher stereochemical 
exigeicy imposed by the polar groupings. Crystal 
anal}^ses ha\^e shown a comparatively small range 
of variation of the lengths and directions of hy- 
drogen bridges. There is in fact a special char- 
acter in hydrogen bridges, which is absent in the 
van der \Vaals type of association. We can pic- 
ture methyl groups so arranged that those from 
one ])rc)tein surface pack into pockets of methyl 
groups from any other protein surface and vice 
Z'crsa. b\' means of a kind of cushioning effect, 
such that the surfaces can be pressed rather closer 
together by means of the spreading of methyl 
grou]:)S, the methyl-methyl distance remaining sub- 
stantially unchanged. But take a hydrogen bridge 
association in which ( say ) 9 or more glutamines 
from a face of one protein interlink with an equal 
number from the face of another protein and the 
situation is quite different. Several arrangements 
of the -CH2- groups, giving larger and smaller 
distances between the faces, might be possible, but 
the transition between the various states would 
not be a continuous one but rather a snapping into 
position. It is this last point which makes clear 
why. in a sense, protein structure systems will 
have pores which allow the passage of molecules 
of appropriately small size even though they are 
not lipoid soluble. Actual attempts to build 
structure systems in which protein units are inter- 
linked by polar side chains will show that it is not 
possible to pack these hedgehog-like units together 
without interstices. There will inevitably be 
lacunae or pores surrounded by rigid boundaries 
of polar groups. In this way we can understand 
the free penetration of water, a phenomenon 
which has always proved difficult to account for. 
when models of membranes involving complete 
monolayers of lipoids are insisted upon. W'ith 
molecules which penetrate through points in the 
framework where the association is of the van der 
Waals type the situation is quite different. In the 



August 16, 1941 ] 



THE COLLECTING NET 



145 



case of cushioning associations of this kind there 
will be no pores of definite size defined by rigid 
atomic boundaries. The penetration of molecules 
b}^ means of nosing their way between non-polar 
groups will thus not be a question primarily of 
size, but far more a question of affinity to the 
groups in the surfaces of the cushions, i.e., of 
lipoid solubility. This picture is oiifered as a pos- 
sible interpretation of the generalization regarding 
permeability of membranes in which Jacobs has 
summed up a large number of his own findings, 
according to which if molecules are sufficiently 
small they will enter the cell regardless of their 
lipoid solubility ; whereas, if they are sufficiently 
lipoid-soluble they will enter the cell regardless 
of their size. 

Even this shght preliminary discussion of per- 
meabilitjr shows that the most significant essential 
points in the picture of structure systems of these 
native protein units is the implication regarding 
the varied nature of the interlinks. Just as the 
specificity of a native protein resides, on this 
theory, in the spatial patterns in which the various 
R-groups are rooted in precise positions in the 
rigid skeleton, so the specificity of a membrane, it 
is suggested, resides partly in these patterns on 
the skeletons and partly in the nature of the inter- 
hnks. With this picture, we can visualize systems 
which have wholly different specificities in virtue 
of different patterns of interlinking, even if there 
is httle or no difference in the specificities of the 
constituent protein units. It follows that great 
differences in permeability by no means imply 
correspondingly great differences in the constitu- 
ents of a membrane or cytoplasmic system. We 
are all familiar with the classical work of Dakin 
and Dudley which showed (T think for the first 
time) that chicken and duck albumin differ in 
their molecular structures, even though no differ- 
ence in chemical composition could be found, a 
finding which points directly to spatial patterns of 
R-groups. As Parpart has pointed out, species 
differences as determined by permeability studies 
are not found to be correlatable with large differ- 
ences in composition. Evidently, as he remarks, 
it is the molecular orientation which is the im- 
portant factor, a suggestion I venture to interpret 
in terms of spatial patterns, not only of R-groups 
but also of interlinks. It is this gradually grow- 
ing conviction among the experts in the field 
which finds in the present picture a fundamentallv 
simple and straightforward interpretation in that 
the specificity of any protein structure system 
whatsoever must necessarily be a function not only 
of the specificities of its constituents but also of 
the specific patterns of the interlinkages. 

The Protein in Modern Dress 

We are then presented with the native protein 
in modern dress, a figure markedly akin, as I shall 



be able to show, to the picture in the minds of 
many Ijiologists during the last eighty years. This 
protein unit is large, Init of definite skeletal struc- 
ture which gives it its protein character. It is 
rigid, so far as the skeleton is concerned, but it 
is covered by flexible R-groups. It may be that, 
so far as its skeleton is concerned, it exists in very 
few varieties of sizes, but each such protein genus, 
covered by this or that selection of the strictly 
limited number of different R-groups, can give 
rise to an astronomically large number of highly 
individualized protein species. Thus we see how 
the protein character can be conjoined with a 
variety sufficient to account for even so many pro- 
teins as would be formed were every suitable 
molecular species injected in turn into every 
species of living animal. Finally the native pro- 
tein, this hedgehog-like structure consisting of 
hard kernel and softer bristles, depends for its 
stability on interlinking with other molecules : it 
cannot exist in isolation. Native proteins will 
hold native proteins by means of groups of bristles 
emerging from definite patches on the kernels and 
so preferentially in certain directions. Since the 
bristles are themselves of varied types and are 
rooted in a definite spatial pattern, whatever this 
may be, it will hold these molecules, on which its 
life depends, in specific fashion. Not any two 
faces of any two proteins will be capable of form- 
ing a stable association, since this depends upon 
the favorable arranffement of not one but many 
R-groups on each face. Cases may be expected 
in which protein faces not adapted to direct inter- 
linking may be able to unite in a structure system 
Ijy using as intermediaries molecules with affini- 
ties to the two faces. The possibility exists that 
the considerable complement of cephalin known to 
be present in erythroc}'te s_\'Stems and thought by 
many experts to be integral parts of the mem- 
brane structure may be playing just such a role 
as this, in view of the groups, both polar and non- 
polar, which are present in these molecules. 

What then can we do with this native protein 
unit, if we wish to build structures possibly rele- 
vant to the nature of cytoplasm and cytoplasmic 
membranes? The answer is that we can build 
protein structure systems of a wide variety of 
types — wide enough I think to provide a starting 
point for a new cxperiuicntal attack on the struc- 
ture of cytoplasm and cytoplasmic membranes in 
general. 

Thus we may start in the obvious way by con- 
sidering systems of the three types, linear, area! 
and volume. A set of native protein united in a 
one dimensional array we shall call a molecular 
fibril, open when it is like the links of a chain, 
closed when it is like the beads in a necklace. 
(A molecular fibril or chain, which may be sug- 
gestive as a hypothesis in a variety of physiologi- 
(Continued on page 148) 



146 



THE COLLECTING NET 



[ Vol. XVI, No. 145 



The Collecting Net 

A weekly publication devoted to the scientific work 
at marine biological laboratories. 

Edited by Ware Cattell with the assistance of 
Boris I. Gorokhoff and Judy Woodring. 

Entered as second-class matter, July 11, 1935, at 
the U. S. Post Office at Woods Hole, Massachusetts, 
under the Act of March 3, 1879, and re-entered, 
July 23, 1938. 

BIOLOGY AT THE UNIVERSITY OF TEXAS 

The school year 1940-41 has been an eventful 
one for biological departments at the University 
of Texas. 

Dr. T. S. Painter, professor of zoology and in- 
ternationally known investigator on salivary gland 
chromosomes, was named Distinguished Profes- 
sor by the University Board of Regents, in line 
with a recently adopted policy of giving recogni- 
tion and increased remuneration to outstanding 
nicml^ers of the regular staff. Dr. Painter is a 
member of the National Academy of Sciences and 
five other American scientific societies, has a long 
list of publications, and was one-time holder of 
the academy's Daniel Giraud Elliot Medal. He is 
starred in American Men of Science. 

In January the botany and bacteriology depart- 
ment held its second annual statewide seven-day 
short course for health officers and laboratory 
technicians, under joint sponsorship with the 
State Department of Health. A week's work in 
L^niversity laboratories was opened to staff" men 
from hospitals, clinics and county health units. 

The Clayton Foundation of Houston awarded 
tliis year a $45,000 grant to the University to 
make possible an extensive study of the presence 
and behavior of vitamins in all kinds of living 
tissue. Dr. Roger J. Williams. University bio- 
chemist and di.scoverer of the vitamin, pantothenic 
acid, was placed in charge of the work. Named 
to assist Dr. Williams are Dr. Maxwell Pollack, 
formerly at Northwestern University and later 
employed by the Pittsburg Plate Glass Company 
of Barberton. Ohio, for research in synthetic 
resins ; and Dr. Alfred Taylor, formerly assistant 
professor of zoology at Oregon State College. 

The Clayton Foundation also made an exten- 
sive grant to establish a new laboratory to study 
brucellosis, a recurrent fever which blights dairy 
cattle, under direction of Dr. Vernon T. Schu- 
hardt. University bacteriologist. Two Eastern 
experts are employed to assist Dr. Schuhardt — 
Dr. Norman B. AlcCuIlough of Detroit, Mich., 
and Leo Dick of Marshfield, Wise, both former 
employees of Parke, Davis & Company. 

A $34,520 grant has been received from the 
Rockefeller Foundation for three years of genetics 
research, under direction of Dr. J. T. Patterson, 
professor of zoology; Dr. Wilson Stone, associ- 
ate professor of zoology ; and Dr. A. B. Griffen, 
research associate in the University's Research 
Institute. 



ADDITIONAL INVESTIGATORS 

Burt. R. L. jr. res. fel. biol. Brown. OM 22. Ka 2. 
Furth, J. assoc. prof. path. Cornell Med. Br 317. 
Lucas, A. M. assoc. prof. zool. Iowa State. OM 29. 
Velick, S. F. res. fel. biochem. Yale. Br 110. 

DATES OF LEAVING OF INVESTIGATORS 

Ballard W. W Aug. 14 Henry, R Aug. 2 

Brooks, M. M Aug. 14 Horn, A. B Aug. 7 

Bullock, T. H Aug. 15 Klotz, J. W Aug. 10 

Costello, D. O Aug. 6 Mitchell, P. H Aug. 2 

Dumm, Mary E Aug. 8 Plough, H. H Aug. 1 

Dytche, Maryon ....Aug. 7 Wolf, E. A Aug. 13 

Forbes, J Aug. 2 Woodward, A Aug. 4 

Oilman, L Aug. 11 Y'ntema, C. L Aug. 13 

Dr. R. Ruggles Gates is rewriting this sum- 
mer at the laboratory his book on "Biological 
Botan}-," which was to have been published by an 
English firm last fall. The typeuxitten manti- 
script, consisting of twenty-five chapters, to- 
gether with drawings for 150 plates, was burned 
when the offices of the publishers were destroyed 
in an air raid on London last Deceinber. The 
handwritten manuscript, from which the typewrit- 
ten one was made, had been mailed back to the 
United States, but was lost in transit. Chapters 
in the l)ook included such topics as chlorophyll, 
])hotosynthesis. stomatal action, rare elements in 
metaiiolism. hormones, vitamins, viruses. X-ray 
patterns in relation to cellular structure, chromo- 
some structure, etc., embodying much recent work. 

Dr. John Blxk, assistant professor of zoology 
at the University of Rochester, with his wife, Mrs. 
Elizabeth Mast Buck, is collecting and making 
experimental observations on fireflies in Jamaica 
this summer. His work is supported by a grant 
from the American Philosophical Society. The 
Bucks will sail from Jamaica on August 21. Dr. 
Gardner Lynn, associate professor of zoology at 
[ohns Hopkins L'niversity, under the auspices of 
the Brooks Fund, is also on the island studying 
the de\eIo]iment of frogs \\-hich have no free- 
swimming larval stage. With these two men are 
John Marbarger and James Dent, both of whom 
took the course in invertebrate zoology last sum- 
mer. The group is working at an elevation of 
about 2.000 feet in the Blue Mountains of Jamai- 
ca in a mansion on a coffee plantation which has 
lieen converted into a laboratory and which has 
Ijeen provided for them by the museum at Kings- 
ton. 



CURRENTS 


IN THE HOLE 


At the following 


hours (Daylight Saving 


Time) the current in the Hole turns to run | 


from Buzzards Bay 


to Vineyard 


Sound: 


Date 


A.M. 


P.M. 


August 17 




12:13 


August 18 


12:50 


1:07 


August 19 ,, - 


1:42 


1:57 


August 20 


2:30 


2:45 


August 21 


3:19 


3:30 


August 22 


4:01 


4:18 


August 23 


4:45 


5:03 



August 16, 1941 ] 



THE COLLECTING NET 



147 



ITEMS OF INTEREST 



At the meeting' of the Corporation of the Ma- 
rine Biological Laboratory on Tuesday Drs. G. 
H. A. Clowes, S. C. Brooks and Columbus IseHn 
were elected trustees of the Laboratory. A re- 
port of the meetings of the Board of Trustees 
and of the Corporation by Dr. Charles Packard 
will be printed in the next issue of The Col- 
lecting Net. 

At the annual meeting of the Woods Hole 
Oceanographic Institution on Thursday, Dr. Van- 
nevar Bush, president of the Carnegie Institution 
of Washington, was elected a Trustee to serve 
until 1944. The six Trustees whose terms ended 
this year were reelected. Among these is Dr. T. 
H. Morgan. 

Dr. Edmltnd V. Cowdry. professor of cytolo- 
gy at the School of Medicine of Washington LTni- 
versity in St. Louis, has been appointed head of 
the Department of Anatomy, succeeding Dr. Rob- 
ert J. Terry. 

Dr. R. W. Gerard has been promoted from 
associate professor to professor of physiology at 
the University of Chicago. 

Dr. N. T. Mattox. who is instructing in the 
Invertebrate course at the Marine Biological Lab- 
oratory, has been promoted from instructor to 
assistant professor of biology at Miami Univer- 
sity, Oxford, Ohio. 

Dr. Carl Herget, who worked at the Marine 
Biological Laboratory two years ago, has been 
appointed instructor in physiology at Cornell 
University Medical College. 

Miss Dorothy Dole, who graduated from 
Bates College in June and who is taking the in- 
vertebrate zoology course at the Marine Biologi- 
cal Laboratory, has been appointed graduate as- 
sistant in zoology at Vassar College. 

M.B.L. TENNIS CLUB 

The annual tournaments of the Tennis Club are 
under way : the finals in some of them are sched- 
uled for this afternoon. 

In the mi.Ked doubles, the finals will see S. 
Lumb and D. Humm playing against L. Te Win- 
kel and D. Lancefield. 

In the men's singles, the following results were 
reached in the second round : Stunkard defeated 
Mavor, 6-2, 6-3 and Evans defeated Williams, 
6-1, 6-1. 

In the women's singles, Mary Chamberlin will 
be one of the finalists, having defeated G. Saun- 
ders, 6-2, 4-6, 6-4. 

In the men's doubles Stunkard and Evans have 
qualified as finalists by defeating Jones and 
Speidel, 6-2, 6-1. 



Dr. Kenneth S. Cole, associate profes.sor of 
physiology at Columbia University, has received 
a Guggenheim fellowship to study the electrical 
aspects of the structure and function of the living 
nerve. 

Dr. George L. Kreezer, assistant professor of 
psychology at Cornell University, has been 
awarded the Snyder grant of $1,000 to continue 
his research on the effect of pre-natal deficiences 
in nutrition upon the development of behavioral 
capacities in the rat. He worked at the Marine 
Biological Laboratory in July. 

Mr. James McInnis, manager of the Supply 
Department of the Marine Biological Laboratory, 
has been appointed chairman of the Falmouth 
Division of the Massachusetts Committee on Pub- 
lic Safety, which has charge of all home defense 
activities. His latest assignment is to organize 
a Gasoline Conservation Committee. 

Dr. C. E. Wilde, Jr., instructor in zoology at 
Dartmouth College, who reported recently for 
work at the Marine Biological Laboratory, was 
drafted shortly after his arrival and is now sta- 
tioned at Camp Edwards. 

Dr. Robert Chambers left on Tuesday morn- 
ing for Cleveland. 

The Eleventh Annual Meeting of the Ameri- 
can Malacological Union will be held at Thomas- 
ton and Rockland, Maine, from August 26 to 29. 

A general scientific meeting will be held Tues- 
day and \\'ednesday, August 26 and 27 in the 
Auditorium for the presentation and discussion of 
work done at the Marine Biological Laboratory 
during the present season. 

At the staff meeting of the Woods Hole Ocean- 
ographic Institution on August 11, William C. 
Derrington of the Fish and Wildlife Service pre- 
sented a paper on "Fluctuations in Fish Popula- 
tion and the Problem of Optimum Yield." 

M.B.L. CLUB 

Tomorrow evening the third of the occasional 
Sunday phonograph record concerts will be pre- 
sented. The opera "Dido and Aeneas" by Henry 
Purcell will be played at 7:15 P. M. in the Fire- 
place Room. 

On Monday, liecause of the lecture in the Audi- 
torium at 8 :00 P. M., the concert will begin at 
9 :00 and will last only forty minutes. The fol- 
lowing numbers will be presented : Sibelius, "Ro- 
mance in C for Strings" ; Sibelius, "Swan of 
Tuonela" ; Sibelius, "Symphony No. 7 in C Ma- 
jor." 



148 



THE COLLECTING NET 



[ Vol. XVI, No. 145 



INVERTEBRATE CLASS NOTES 



A well organized and extensive study of the 
marine annelids together with two long field trips 
made up the scheduled bill of fare for the past 
week. In addition to the dissection of Nereis and 
Arenicola. we tried identifying the different local 
s]3ecies with the help of a key. Through the ef- 
forts of the collecting crew fifty-one of the fifty- 
nine species were available. The large number 
of people working in the lal:> Sunday indicates the 
interest taken in this aspect of the work. 

Though most of a week had elapsed since Dr. 
Rankin's last lecture, a student was recently heard 
laughing explosive!}', having just caught on to 
one of the former's "jokes." The time lag is un- 
derstandable ; "The difference between a bird with 
two wings and a bird with one is just a matter of 
opinion." Explanation supplied on request. 

Our collecting trips to Kettle Cove and Cutty- 
hunk proved very successful, both from the point 
of view of the number of specimens collected and 
of sheer good fun. For there is the two-hour boat 
trip to Cuttyhunk, with group singing, jokes, and 
banter ; the sustaining if brief picnic lunch, and 
the thrill of discovery. As many as 135 speci- 
mens were collected by a single team. The old 
Winijrcd whistled herself hoarse before the last 
team reluctantly gave U]) the chase. 

Saturday night twenty-four invertebrates left 
the dance, boarded row boats, and made for 
Devil's Foot Island. Two trips were required to 
ferrv the entire group. Suj.iper of hamburgers 

PROTEINS 

(Continued fr 

cal problems, is of course a linear iiattern of mole- 
cules. It is to be distinguished clearly from an 
atomic fibril or chain which is a linear pattern of 
atoms, such as have enjoyed for no good reason 
so far as I can see, some vogue in certain quarters 
in diagrams of supposed physiological situations. 
WHien the native protein units are arranged 
in two dimensional formation we shall talk of a 
fabric or skin, which may be open like the sail of 
a ship or a piece of paper, or closed like the sur- 
face of an orange or of a torus or anchor ring. 
A set forming a three dimensional array completes 
the three basic categories. Each type is of course 
flexible, somewhat extensible, has some tensile 
strength. Each type represents a system having 
organization in a definite sense, even if this sense 
is not entirely easy to make perfectly precise. The 
specificity we have talked aliout as characteristic 
of all protein structure systems is present, present 
as before explained in a dual form. There is 
specificity of the R-group patterns on the faces 
of the native protein units which are disengaged 
and there is specificity in the interlinks, derived, 
of course, from the R-group patterns of the faces 



was followed by ever popular group-singing, this 
to be interrupted by peals of thunder. The wind- 
driven rain sent the picnicers scurrying to the 
boats. The trip to the mainland, ordinarily re- 
quiring twenty minutes, required considerably 
more time with overladen boats and opposing 
wind and tide. Indeed, the last group, in spite of 
the capable oarsman, was obliged to abandon the 
crossing. The boys succeeded in dragging the 
boat to Ram Island, where, in the crowded shelter 
of the overturned boat, they awaited a more fa- 
vorable opportunity. After about two hours, the 
eight adventurers once more headed homeward 
only to find they lacked one oarlock. After con- 
triving a substitute from the remains of a chair 
and several shoe-laces, the group arrived at 5 :30 
A.M., to be welcomed by a hardy few, determined 
to miss none of the fun. 

What with the annelid drawings being due 
.Monday evening at seven, a beach party was or- 
ganized to take advantage of the unexpected holi- 
day. A marshmallow roast at Gansett beach 
made for less threat from tide and storm, and 
about as much fun as Saturday's adventure. 

A certain member of our class, now called "the 
ancient marooner," left his girl stranded at Pen- 
zance Point Light when the latter shoved the boat 
into the Woods Hole current. The valiant col- 
lecting crew effected a rescue three hours later. 

— Louise Gross and BUI Batchclor 

IN ACTION 

om page 1-15) 

which are interlinked. 

It need hardly be pointed out that combinations 
of these three basic types will give compound fib- 
rils of any cross section, compound fabrics of any 
thickness, and so on. Since it is essential to pay 
special attention to the orders of magnitude in- 
volved, it should be noticed that a fibril of length 
ten microns will contain something of the order of 
3000 units the size of insulin and that for a cross 
section of one tenth of a micron in diameter we 
shall need some thirty strings of single units ar- 
ranged in parallel. A skin, one molecule thick, 
will contain say 5 X 10* units the size of insulin 
per square micron, so that a protein complement 
of the order of 10~^- mg. per square micron as 
found in the red cells of many species can corre- 
spond to a fabric, one molecule or at most a few 
molecules thick. 

In the case of cytoplasm, structure systems 
within such a membrane can be devised forming 
one and the same system with the membrane, 
having as high a proportion of water to protein 
as we wish. Thus the structure systems already 
devised (Cold Spring Harbor Symposium. 1941) 



August 16, 1941 ] 



THE COLLECTING NET 



149 



in which long molecular fihrils are joined in (say) 
four-way native protein units (topologically of the 
ty]5e of the diamond structure) give water com- 
plements of 70 to 99 per cent, corresponding to 
fihrils of lengths say 100 A to 0.7 microns. Evi- 
dently with such structure systems even the nu- 
torious jellyfish, with its immense water comple- 
ment, can be accommodated. 

It is not possible to demonstrate in detail on 
this occasion all the ways in which these simple 
types of structure system suggest interpretations 
of many facts regarding cytoplasm and membrane 
behavior. It seems of importance to call attention 
to the localization of different interlink types in 
different parts of the membrane structures, which 
explains why permeability properties can be modi- 
fied by fewer molecules than are required to cover 
the surface with a layer one molecule thick. Per- 
meability to certain polar molecules will evidently, 
on this model, be profoundly modified in the pres- 
ence of suitable molecules only at certain interlink 
regions of specific nature where such molecules 
normally enter. Size is of importance, we notice, 
specially with respect to molecules entering the 
rather definite pores, outlined by polar groups rig- 
idly held in hydrogen bridges. Size will be less 
important in the case of the penetration by the 
comparatively non-polar molecules whose natural 
entry points will be the hydrocarbon interlink re- 
gions, in which suitable groups cushion against 
one another, no definite pores of strictly defined 
size being present. 

Among a great variety of relevant considera- 
tions, the fundamental one appears once more to 
relate to specificity. The bewildering variety of 
red cell memliranes is here seen to be compatible 
even with a great economy of native proteins. 
Thus, even if, as some have suggested, there is a 
considerable similarity in the proteins involved, 
exploitation of the many ways in which the same 
units can form structures of utterly different spe- 
cificities by different patterns of interlinkings will 
provide sufficient variety to cover all the facts so 
far elicited. As an example of the way in which 
specific permeabilities may be dependent upon 
quite minor and in any case strictly localized R- 
group situations a possible correlation between 
permeability to urea and the presence of the car- 
boxylic acid groups in the form of glutamine and 
aspargine may perhaps be suggested. May it not 
be possible that reptiles differ from the birds in 
some such way? As an example of ways in 
which local specificities of membranes may have a 
far reaching influence on structure systems, we 
would call attention to the possibility that the hy- 
drophilic R-group patterns on the outside of a 
plant plasma membrane may determine the pat- 
tern in which cellulose is laid down. 



The Protein in Modern Dress 

In summing up, I should like to come back to 
my starting point. The native protein unit in 
modern dress, the true megamolecule, is essential- 
ly a structure-forming substance ; as such it ful- 
fils the essential requirement of the biologist. Its 
capacity to form structures, we have seen, allows 
us to picture systems of immense size and im- 
mense chemical complexity in which great flexi- 
bility is conjoined with rigid foci. Nevertheless 
such systems are built in essence upon simple 
themes. Specificity of the units implies specificity 
also of interlinks and it is this higher order of 
specificity which, I would suggest, is the true ex- 
planation of organization which has been postu- 
lated by biologists for so long. "We must," said 
Briicke writing in 1861, "attribute to living cells, 
beyond the molecular structure of the organic 
components which they contain, still another 
structure of a different type of complication. It is 
this which we call by the name of organization." 
Again in the writings of Wilson we read, "We 
assume as our fundamental working hypothesis 
that the specificity of each kind of cell depends es- 
sentially upon what we call its organization. . . . 
We cannot, it is true, say precisely what organiza- 
tion is, but we can hardly think of it as other than 
some kind of material configuration of the protein 
substance and one that involves a differentiation 
of parts and their integration to form a whole, as 
Herbert Spencer long since urged." Into all these 
statements we can, without forcing, read the new- 
est picture of the native protein, which is thus seen 
to be the direct descendent and the present-day 
embodiment of Pfliiger's living molecule, postu- 
lated in 1875 and Verworn's biogen. Perhaps 
the clearest exposition of this point of view is the 
statement which occurs in the course of Whit- 
man's celebrated attack on the cell theory de- 
livered as an Evening Lecture at this Laboratory 
in 1893 and published in the first volume of "Bio- 
logical Lectures." Leaving the accepted cell 
theory in rags and tatters behind him, the first 
Director of the Laboratory proceeds to discuss 
what can be the nature of the "formative process- 
es" in living matter. "The answer to our ques- 
tion," he says, "may be difficult to find, but we 
may be quite certain that when found it will rec- 
ognize the regenerative and formative power as 
one and the same thing throughout the organic 
world. It will find ... a common basis for every 
grade of organization and it will abolish those fic- 
titious distinctions we are accustomed to make be- 
tween the formative processes of the unicellular 
and multicellular organisms. It will find the secret 
of organization, growth, development, not in cell- 
formation, but in those ultimate elements of living 
matter, for which idiosomes seems to me an ap- 
propriate name. . . What these idiosomes are, and 



150 



THE COLLECTING NET 



[ Vol. XVI, No. 145 



how they determine organization, form and differ- 
entiation, is the problem of problems on which we 
must wait for more light. All growth, assimila- 
tion, reproduction and regeneration may be sup- 
])osed to have their seat in these fundamental ele- 
ments. They make up all living matter, are the 
liearers of heredity and the real builders of the 
urganism." 

Logic and History 

In this statement, attributing to the protein not 
only a dominant but the preeminent role in bio- 
logical structures. Whitman perhaps goes further 
than many, aware of the great importance of lip- 
ids, carbohydrates and other molecules, would be 
willing to go today. But especially in view of the 
protein crisis at the present time, I for one would 
wish to give great weight to the considered judg- 
ments of practicing biologists, who in their wide 
ex]ierimental studies may perhaps get from the 
feel of the material a truer view, even if it be 
partly intuitive, than those who lack such intimate 
contacts with protein in its living state. After all, 
it is (unhappily) only the biologist who has a 
special stake in the attack on the living protein : 
alone aware of all the challenges that a living or- 
ganism can make, he must decide on the adequacy 
of theories of ])rotein structure in the last resort. 

To my way of thinking, the idea in the mind 
uf Whitman and many others such as Fletcher 
who preceded him, when translated into modern 
dress, finds a ready interpretation in the picture 
of the native protein drawn from the work of 
Sxeclberg, the work of the immunologists. the work 
of the protein crystallographers, which finds also 
generalized and precise expression and confirma- 
tion in modern and classical geometry. The native 
protein, the megamolecule. the most specific of all 
substances, is a structure-forming unit whose sta- 
bility indeed depends upon its state of association. 
Such stabilization is accomplished, apparently, in 
the mixed structure systems in which protein. 
Water and other substances are organized with 
superspecificity. But the essence of all such struc- 



ture systems resides, so far as I can see, in one 
thing, namely the maintenance intact of the native 
protein structure. In saying this, I am, of course, 
only saying what all workers with living material 
have always known, at least in their unconscious 
minds. What, then, is this essential structure, 
upon which, if these workers have read the 
facts aright, life itself depends? I suggest it 
is the closed fabric structure, a distinctive 
structural type (as Wu has shown is absolutely 
necessary) which is potentially immortal as it 
passes from generation to generation through the 
ages and yet is of the utmost lability and can 
l)reak down in myriad ways. The general tenor 
of one's thinking, perhaps also of one's physiologi- 
cal processes, will no doubt play a large part in 
determining for each of us whether the idea of 
molecular structures in the form of closed atomic 
fabrics, carrying substituents rooted in spatial pat- 
terns on the surface, appears fantastic and ob- 
scure, as it has J^een adjudged by some, or emi- 
nently reasonable and straightforward, as it has 
been adjudged by others. Even were the major- 
ity reaction of the former type, we need hardly 
be alarmed. Such matters have their relevance 
in the history of Science, but none in the logic of 
Science. It has happened more than once that an 
idea, ultimately useful as a tool in subduing the 
seeming disorder of the external world, did not at 
first sight appear sober and reasonable to all the 
most distinguished contemporary thinkers. To 
estalilish its right to a place in the sun, an idea 
requires no universal assent. To be useful, it is 
sufficient that some should foster its development 
l)y attempting to understand it and that others 
should study its implications by experiment and 
ol)servation and reflexion. Now that its century- 
long period of gestation is over and its embryonic 
stages are nearing completion, we await with con- 
fidence not unmixed with impatience the serious 
study of its implications in biological structure 
jjroblems. 

(This article is based upon a lecture delivered at 
the Marine Biological Laboratory on July 25.) 



INTERPRETATIONS OF EFFECTS OF CO AND CN ON OXIDATIONS IN CELLS 

(Continued from page 137) 



dift'erent eft'ects upon the rate of O- consumiition 
were i)roduced. depending upon the stage of de- 
velopment. The results were as follows : 

Methylene blue produced a rate of 130% in the 
unfertilized stage of Arhacia as compared with the 
control; ISO^f in the first cleavage stage; a slight 
increase in the jiluteus and either a decrease or no 
effect in the gastrula stage. 

Carbon monoxide produced a slight decrease to 
about 92% of the controls in unfertilized eggs; 
a considerable decrease to about 20% of the nor- 
mal in the first cleavage stages, after which the 



decrease was less, so that in the late gastrula 
stage, the rate was 50%. In all of these cases the 
addition of methylene blue increased the rate 
about 10%. 

KCN caused a varying degree of decrease. For 
example in Astcrias eggs, unfertilized, the rate 
was 63% of the control; while in the blastula 
stage of Arbacia it was 25% and in the late gas- 
trula stage of Arbacia it was 6%. 

These experiments suggest new aspects of the 
respiratory enzymes and the redox systems direct- 



August 16, 1941 ] 



THE COLLECTING NET 



151 



ly associated with them. In living cells methy- 
lene blue is about half reduced and half oxidized 
(the ratio of the sum of the reductants over the 
sum of the oxidants, 



[Red] 
[Ox] 



50 



50 



so that its maximum poising action is available. 
Any system has its maximum electron transfer- 
ence as the potential changes when the ratio of 
oxidants to reductants is one. Since methylene 
blue induces different changes in the rate of O2 
consumption at dififerent stages of development of 
the embryo, one may well inquire whether the 
optimum redox potential at which the respiratory 
enzyme and its associates function is dififerent in 
dififerent stages of the same animal. If the redox 
potentials are different, then the enz)'me systems 
must be different or they must assume different 
roles at different stages of development. 

The above interpretation would also explain 
the differences in cyanide-sensitivity as found in 
different types of living cells. Since cyanide is 
known to produce a negative redox potential, 
those respiratory systems which normally have a 
50/50 ratio at the redox potential approximating 
that produced by cyanide, would be less affected 
than systems whose potentials are farther re- 
moved. The former would be known as '"cyan- 
ide-insensitive" systems and the latter as "cyan- 
ide-sensitive". In other words, if the ratio of the 
sum of the oxidants over the sum of the reduc- 
tants was changed from 50/50 to 5/95 by KCN, 
there would be present a greater concentration of 
reductants which could not reverse back and forth 
between oxidized and reduced forms. This would 
produce a slower rate. Furthermore, if the redox 
potential of the systems were not changed by 
cyanide, then no change in the rate would occur. 
This theory eliminates the necessity of trying to 
explain the action of cyanide by assuming that 
there is an affinity with cyanide in one case and 
none in other cases. This theory amplifies a 
former hypothesis of the writer on the action of 
cyanide in living cells, namely, that there is no 
combination with the respiratory enzyme, but 
merely a lowered redox potential, "freezing" in 
the bivalent form a large concentration of elec- 
trons of the Fe of the heme radical which would 
otherwise be able to shift reversibly back and 
forth from the bivalent to the trivalent form. This 
shift, Fe++ ^ Fe+ + + , which must occur for 
respiration, remains mainly in the Fe++ state, so 
that the concomitant transfer of activated hydro- 
gen to oxygen, which is the object of respiration, 
cannot take place. 

In the case of methylene blue, the same princi- 
■ pies may be applied. If methylene blue raises the 



redox potential, for example, of a comj^onent of a 
respiratory system so that its ratio of o.xidants to 
reductants is 50/50, the rate of respiration should 
he increased. If however the system affected has 
its equilibrium ratios changed by the addition of 
methylene l)lue so that the concentration of reduc- 
tants is greater than that of oxidants, or vice 
versa, then there should be a decrease in the rate. 

This tentative explanation is offered of the ef- 
fects of these accelerators and inhibitors on O2 
consumption. It is difficult to account for the ef- 
fect of CO, because so far, no stable potentials 
have been found in the presence of CO with the 
methods in use. 

Theorell's (Publications A.A.A.S. ?^14, 1940, 
p. 136) conclusions on cytochrome C which also 
has a heme radical as its active group and a high 
redox potential, are of interest in this connection. 
He states that at physiological pH values, i.e. 
around neutrality, there are amino residues at- 
tached to the Fe of the heme radical which pre- 
vent it from forming additional compounds with 
cyanide or CO. Only in alkaline solutions does 
ferricytochrome-C form a complex with cyanide ; 
and only in acid and alkaline solutions does ferro- 
cytochrome-C form complexes with CO. These 
explanations may well apply to the respiratory 
enzyme although nothing so definite is at present 
known. The experiments of Clark (Cold Spring 
Harbor Monographs. VII. 18, 1939) and those of 
Barron (lour. Biol. Client. 121, 285, 1937) on 
hemochromogens and cyanide were done at high 
pH values up to pH 13. Their results may well 
fit in with the above theory of Theorell when al- 
kaline solutions are used, but should not be used 
to explain what takes place at pH 7.0 in living 
cells. 

II. Effects on Cleavage. 

A few of the observations on cleavage are as 
follows : KCN produced multiple aster formation 
without cell division even in the presence of meth- 
ylene blue at this concentration. The blastula 
stage of starfish remained alive 8 days without 
further development. CO produced cytolysis but . 
methylene blue protected the embryo or tgg from 
cytolysis. 

Development of the embryo took place after 
the removal of CO. However development in 
the blastula stage of Arbacia was stopped even 
when CO was replaced by air. Other stages were 
in the main not permanently affected by CO. 
Metln'lene blue has doubled the life of both Ar- 
bacia and Astcrias ; it ]M-oduced faster develop- 
ment, it increased the size of the plutei in Arbacia 
from 280 fx. an average size in the controls, to 
420 /x in length. 

(This article is based upon a seminar i-eport pre- 
sented at the Marine Biological Laboratory on 
August 5.) 



152 



THE COLLECTING NET 



[ Vol. XVI, No. 145 



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August 16, 1941 ] 



THE COLLECTING NET 



153 




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154 



THE COLLECTING NET 



[ Vol. XVL No. 145 



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THE COLLECTING NET 



155 




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156 



THE COLLECTING NET 



[ Vol. XVL No. 145 




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Vol. XVI, No. 9 



SATURDAY, AUGUST 23, 1941 



Annual Sub.=icription, $2.00 
Single Copies, 30 Cents. 



THE UTILIZATION OF AMMONIA BY 
CHILOMONAS PARAMECIUM 

Dr. Jopin O. Hutciiens 
The Joliiis Hopkins University 

Cliiloinoiias paranicciuui is a cryptomonad flag- 
ellate about 30 micra long and 15 micra wide 
which was found bv Alast and Pace (Prof o plas- 
ma, Bd. 29, S. 326-359, 1933^. 

and subsequently lay others to 
grow in a variety of solutions 
using single amino acids, urea 
or aminonia as a source of ni- 
trogen, and lactate, acetate, 
formate, or CO2 as a source 
of carbon. Table I (Solution 
A) shows the composition of 
one of the solutions used b}' 
Mast and Pace and used by 
me in earlier work (Hutchens, 
/. Cell, and Com p. Plivsio!.. 
V. 17, pp. 321-332, 1941). It 
will be seen that ammonia is 
the sole source of nitrogen, 
and that acetate is the only 
source of carbon. In such 
solutions the chilomonads at- 
tain a population of only 5-6 
thousand/cc. and die on the 
fourth and fifth days of the 
culture. They can be subcultured indefinitely, 
however, if transfers are made on the second or 
third days. {Continued on page 170) 



M. IS. IC. €alcn^c^r 

TUESDAY, August 26, 9:00 A.M. 
General Scientific Meeting 

Continued at 2:00 P. M.' 

WEDNESDAY, Aug. 28, 9:00 A.M. 
General Scientific Meeting 

WEDNESDAY, Aug. 28, 8:00 P.M. 
Motion pictures of local fauna and 
flora, George G. Lower. 

THURSDAY, August 29, 8:00 P.M. 

Lecture: Dr. Milton Levy: "Chem- 
ical Studies of the Chick Em- 
bryo." 

FRIDAY, August 30, 8:00 P.M. 
Lecture: Dr. Rudolf Schoenheimer: 
"The Dynamic State of the Body 
Constituents." 



THE OFFICIAL MEETINGS OF THE 
MARINE BIOLOGICAL LABORATORY 

Dr. Charles Packard 
Director 

The meetings of the Corporation and Board of 
Trustees of the Marine Biological Laboratory, 
held on August 12, were uneventful in the sense 

that no controversial matters 

cai::e up for discussion. The 
Officers and Trustees eligible 
for re-election were voted in 
by unanimous consent, as were 
also the three new Trustees 
and the fourteen ne^v members 
of the Corporation. Dr. W. J. 
V. Osterhout was elected a 
Trustee Emeritus. The Treas- 
urer, Mr. Riggs, reported that 
the Laboratory is free of all 
indelitedness and that its fi- 
na'"ces are in good condition. 
The numl)er of investigators 
and students in attendance this 
summer is only slightly below 
the average of the past five 
years, which is 485, though 
considerably below that of 
1937 and 1940. In those sea- 
sons, more than 500 were reg- 
istered, and the Laboratory was uncomfortably 
crowded. We may congratulate ourselves that in 
spite of wars and rumors of wars, investigation 



TABLE OF CONTENTS 



The Official Meetings of the Marine Biological 
cal Laboratory, Dr. Charles Packard 15V 

The Utilization of Ammonia by Chilomonas 
Paramecium, Dr. John O. Hutchens 157 

Comparison of the Respiratory Rates of Dif- 
ferent Regions of the Chick Blastoderm 
During Errlv Stages of Development, Dr. 
F. S. Philips 159 

Studies on Conditions Affecting the Survival 
in vitro of a Malarial Parasite (Plasmo- 
dium lophurae), Dr. W. Trager 162 



163 



164 



The Effect of Dyes on Response to Light in 
Peranema trichophorum. Dr. C. Hassett.... 
Program of the Summer Meetings of the Gen- 
etics Society of America at Cold Spring 

Hfibor, Long Island, N. Y 

Summer Activities of University of Chicago 

Biologists 166 

Items of Interest 167 

Invertebrate Class Notes 168 

Some Recent Books in the Biological Sciences 168 



§ 


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s 


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August 23, 1941 ] 



THE COLLECTING NET 



159 



and instruction have gone on almost unhampered. 

The comjiletion of the new wing of the Lihrary 
marks another stage in the development of this 
important part of our institution. In earlier days 
it was housed on the Protozoology Lalioratory : 
then in Room 217 of the Crane Building, which 
soon proved inadequate. When the Brick Build- 
ing was erected, the shelf space allotted to the 
Library was deemed sufficient to accommodate the 
growth of many years. But the rapid increase in 
the number of back sets, current periodicals, and 
separates, made imperative a very substantial ad- 
dition to the stack room. This we now have, 
thanks to the generosity of the Rockefeller Foun- 
dation. The excellent arrangement of the journals 
and the adequate provision for readers make the 
Library a most attractive place to the investigator. 

During the past year, the storage battery, which 
was damaged by the flood of 1938, was disman- 
tled. This means that we now have no source of 
light and power other than the town supply. If 
this fails, as it has on more than one occasion 
during the present season, our pump stops, and 
our supply of running sea water is rapidly de- 
pleted. Now sea water is our life-blood ; without 
it we could not continue our observations and ex- 
periments for a single day. It is necessary to 
keep the storage tank full so that if the pump 
stops temporarily there may be enough water to 
supply our needs until the electric current is re- 
stored. Only by using sea water sparingly can 
this be done. Investigators can ensure the con- 
tinuance of their experiments by using a minimum 
of sea water. 

The Supply Department has made two changes 
in the method of collecting and keeping certain 
kinds of live material. It no longer uses its own 
fish trap, but relies on the catch taken in the much 
larger nets of local fishermen. This plan has 
worked out very well this season. Urchins and 
stars, dredged from the sea bottom, are either de- 
livered to investigators directly from the boat, or 



placed in shallow tanks indoors, where they ap- 
parently keep in better condition than they did in 
the wire cages under the floats. 

Dr. Pond called attention to the fact that many 
items essential to experimental work are becoming 
increasingly difficult to secure. For example, 
copper, brass, tin, ruljber and aluminum can be 
obtained only in odd lots, if at all. Acetic, citric 
and carbolic acid, solvents and many other chem- 
icals are in the same class. Fortunately ethyl al- 
cohol can be purchased, though some restrictions 
are placed on it. Every effort will be made to 
obtain these supplies, but obviously investigators 
can be furnished only very limited amounts. 

The relation of the Laboratory to the present 
emergency was summarized by Dr. Lillie in the 
following vvords : 

'"During the present international and national 
crisis it is both the privilege and duty of educa- 
tional and research institutions to fulfill their nor- 
mal functions as effectively as possible. We have 
done so this season and will continue so to act. It 
is not necessary for me to emphasize to the mem- 
bers of the Marine Biological Laboratory the 
value and dignity of our role in the advancement 
of science and education, nor, I am sure, the re- 
sponsibility that rests upon us to defend and pre- 
serve our institution. 

"We shall, of course, stand ready to make sac- 
rifices in the interests of national defense if called 
upon to do so. Our location might make the use 
of some of our facilities for inshore patrol work 
valuable to the Navy Department ; I understand 
that there are plenty of rumors about, that they 
may be requisitioned in whole or in part. We 
have had no official notification that this is likely 
to be the case, and we feel assured that our own 
interests will be respected as far as possible in any 
demand that might be made. It is my ow-n opin- 
ion that it is improbable that any such demanri 
will be made this year, but I have no opinion 
about the subject beyond that time." 



COMPARISON OF THE RESPIRATORY RATES OF DIFFERENT REGIONS OF THE 
CHICK BLASTODERM DURING EARLY STAGES OF DEVELOPMENT 

Dr. Frederick S. Philips 

Osborn Zoological Laboratory, Yale University ; Department of Physiological Cheiuistry, 

Yale University, for the academic year 1941-1942 



The consideration of the regional differences in 
embryonic processes which exist in the head- 
process embryo of the chick, or for that matter in 
any embryo, demands an analysis of the nature of 
the biochemical phenomena which must be in- 



volved in such morphogenetic activity. A few in- 
vestigators have already attempted the beginnings 
of this sort of analysis in the chick. Rulon, 1935. 
and Miller, 1941, have both found that the vital 
dye, Janus green, under anaerobic conditions is 



The Collecting Net was entered ns second-class matter July 11, 193,5, at the Post Office at Woods Hole. Mass., 
under the Act of March 3, 1879, and was re-entered on July 23, 1938. It is devoted to the scientific work at 
marine biological laboratories. It is published weekly for ten weeks between July 1 and Sejitember 15 from Woods 
Hole, and is printed at The Darwin Press, New Bedford, Mass. Its editorial offices are situated in Woods Hole. 
Mass. Single copies, 30c by mail; subscription, $2.00. 



160 



THE COLLECTING NET 



[ Vol. XVI, No. 146 



most rapidly reduced in the region of the embryo 
which contains Hensen's node. Jacobson, 1938, 
has reported a diminution of glycogen and an in- 
crease in lipoid material in epiblast cells which 
are invaginating in the primitive streak. The 
present work is a study of the rate of oxygen con- 
sumption in different regions of the head-process 
embryo in order to investigate the existence of 
possible correlations betvveen one phase of respir- 
atory metabolism and the various regional differ- 
entiations in embryonic activity previously de- 
scribed. 

A second point of interest in the present work 
arises out of the recently published observation 
( Philips, 1941 ) that there is an increase in the 
rate of oxygen consumption of the whole chick 
blastoderm during the first four hours of incuba- 
tion. In extension of this earlier observation the 
present investigation considers the question of 
whether a resjjiratory enhancement takes place in 
the area pellucida of the blastoderm during this 
same early period. It also considers the changes 
in rate of oxygen consumption of the embryonic 
material up to the time of the formation of the 
head-process embryo. 

After a preliminary period of incubation at 
38°C. blastoderms were removed from the yolks 
of eggs ( ^Vhite Leghorns and in a few cases. 
White Rocks ) in Ringer-buffer solution. Dissec- 
tions of the embryonic regions were carried out 
with fine glass and steel needles and the area of 
the isolated pieces was measured by means of an 
ocular micrometer. Oxygen consumption was de- 
termined in the Cartesian diver microrespirometer 
of Linderstrom-Lang with the modifications intro- 
duced by Needham, Boell, and coworkers. The 
volume of the divers used was about 30 c. mm. 
with rate constants of about 20 m. fi\. O- con- 
sumed for every centimeter change of the levels 
of the Brodie fluid. The embryonic tissues after 
isolation from the embryo were placed in the diver 
bulbs with 2 c. mm. of Ringer-buffer containing 
0.4 per cent glucose. A stream of oxygen was 
passed through the divers before placing the al- 
kali seals in their necks. After the oxygen con- 
sum]ition had been followed for about two hours, 
the pieces were removed from the divers and their 
total-nitrogen determined. The nitrogen estima- 
tion was made by a modification of the micro- 
Kjeldahl method of Needham and Boell, 1939. 
Dr. Boell, who plans to pijDlish a description of 
this modification in the near future, has found it 
reliable between 0.5 and 25 y N. Since the value 
of the total-nitrogen was known, the Qoo' of the 
embryonic pieces could be calculated as the 
m. (111. Oo consumed/hoiuVy N. 

In order to test the reliability of the described 
techniques for the present analysis the rate of 
oxygen consumption was compared in right and 
left halves of the area pellucida of the same head- 



process blastoderm. The halves were produced 
by a median cut through the head-process and 
primitive streak of the isolated area pellucida. 
Each half contained about 2 y total-nitrogen and 
consumed about 0.25 c. mm. Oo in two hours. 
The ratio of the rate of the right half to that of 
the left half was 1.02 according to the average of 
results from twenty determinations. Moreover, 
the average Qoo' of 24 right halves and 28 left 
halves was in both instances 78. Since there is 
such a close agreement in the value for the two 
halves, it appears that the techniques used for the 
present determinations give reliable results. It 
may also be concluded that the rate of oxygen 
consumption under the conditions of the present 
experiments is similar in both halves of the area 
pellucida of the head-process embryo. 

The next observations concern the study of the 
different regions of the head-process pellucida 
area. The head-process embryos used varied in 
development from the early head-process to the 
early head-fold stages. The area pellucida was 
divided into six pieces containing respectively the 
head-process, node and anterior streak, middle 
streak, posterior streak, and right and left lateral 
regions. In order to obtain sufficient material for 
the nitrogen analyses the same regions from two 
or three blastoderms were analysed together in the 
same divers and Kjeldahl flasks. Two or three 
pieces of the node and anterior streak region con- 
tained about 2 y N. 

In Table I it can be seen that the head-process, 
node and anterior streak, and middle streak re- 
gions have very similar Oo.,' values. The other 
regions have somewhat higher rates, but from the 
values of the standard deviations it does not ap- 
]5ear that the differences are significant. In a few 
determinations of the latter regions the amounts 
of material were small enough to give serious er- 
rors in the micro-Kjeldahl determination used. It 
is also probable that in these determinations the 
total nitrogen values were incorrect due to losses 
of tissue material during the transferring proce- 
dures which would result in rate values that were 
too high. However if the small differences noted 
are real, they may be of considerable importance 
in the regional differentiation of the head-process 
embryo. 

The results of the observations on the Qoo' of 
the area pellucida from the stage of the unincu- 
bated blastoderm to the 37-hour embryo (11-12 
somites) show that there is an increase in the rate 
during the first 17 hours of development and then 
the rate remains similar through the latest stage 
determined. For these determinations as much of 
the area pellucida was used as possible. The Qoj' 
of the tissue in the unincubated blastoderm was 
about 33. By the definitive streak stage the value 
had increased to about 75. Similarly the values 
for the head-process, 5-6 somite, and 11-12 somite 



August 23, 1941 ] 



THE COLLECTING NET 



161 



Table L 

The Qo/ (m. /* 1. O2 consmned/hour/y N.) of different regions of the area pellucida during 
the first hour of measurement. Averages of 10 determinations are given with the standard deviations. 



Head- 
Process 


Node and An- 
terior Streak 


Middle 
Streak 


Posterior 
Streak 


Right 
Lateral 


Left 
Lateral 


77 

± 9.5 


75 

± 7.3 


74 

± 10.8 


85 

± 13.5 


89 

± 19.8 


92 

± 19.9 



stages were respectively 81, 75, and 78. These 
latter values may be converted into dry weight 
Qo, rates by dividing them by the factor 9. (This 
latter factor is based on the observation that the 
dry weight of chick embryos older than 4-days 
contains about 1 1 per cent total-nitrogen. ) It 
then can be seen that the present observations 
give values for the stages between 17 hours and 
37 hours of development which fall between 8.3 
and 9.0 c. mm. O2 consumed/hour/mg. dry 
weight. Using the Fenn respirometer similar 
values have been found for stages ranging from 
the 1-day area pellucida to the 3-day embryo, 
namely 9.2 to 12.1 (Philips, 1941). With the use 
of the Warburg apparatus Romanoff has shown 
in unpublished experiments that from the 4-day 
embryo to the 7-day embryo the Qo., also remains 
at a similar level, between 9.0 and 10.5. These 
data indicate that the level of the rate of oxygen 
consumption of the whole area pellucida and the 
embryo remains relatively constant between the 
definitive streak stage and the seventh day of de- 
velopment. 

The Qo.,' of the area pellucida of the unincu- 
iDated blastoderm is of the same magnitude as the 
value for the whole lilastoderm at this stage 
(Philips, 1941). Similarly the increase in rate 
of the area pellucida during the first four hours 
of incubation is like that found in the whole blas- 
toderm. The continued increases in rate of the 
area pellucida up to the seventeenth hour of de- 
velopment have a possible explanation. There ap- 
pears to be very little change in the total nitro- 
geneous material contained in the whole area pel- 
lucida during this early period of development 
when the cell-number is being tremendously in- 
creased. Thus this region contains about 3 y N 
in the unincubated blastoderm and about 3.9 y N 
in the head-process embryo. Accompanying the 
increasing cell-number there is, however, a 
marked increase in the total amount of oxygen 
consumed/hour, from about 100 m. fx\. in the 
pellucida area of the unincubated blastoderm to 
about 313 m. ;ul. in the head-process stage. It, 
therefore, appears that during the first 17 hours 



of development nitrogenous, as well as other ma- 
terials, are being converted from inert, storage 
elements in the form of yolk granules into actively 
metabolizing cellular constituents. In subsequent 
stages of development the increase in the total ma- 
terials of the area pellucida and embryo more or 
less keeps pace with the increasing cell-number 
and oxygen consumption. 

One of the stated objects of the present work 
was the investigation of possible correlations be- 
tween differences in the rate of oxygen consump- 
tion of the various regions of the head-process 
embryo and the differences in embryonic activity 
known to exist in these regions. Under the spe- 
cific conditions of the present experiments no re- 
gional differences of any significance have been 
observed with regard to the rate of cellular oxida- 
tions. However, caution must necessarily be ob- 
served in relating this condition to the various 
regionally differentiated embryonic activities of 
the head-process embryo. It is entirely possible 
that the embryonic activities may be suspended 
on removal of the regions from the embryo and 
during the measurement of their respiration m 
vitro. Nevertheless, with this possibility in mind 
it is apparent that these results do not support 
Child's axial gradient theory of development. No 
gradient in the rate of cellular oxidations has been 
found where according to the theory it ought to 
exist. Furthermore, on the basis of the theory- 
one might expect that embryonic stages as widely 
different in embryonic activity as the definitive 
streak, head-process, and 7-day embryo would 
possess widely differing rates of oxygen consump- 
tion. This is not the case. 

There still remains the apparent discrepancy 
between the results of Rulon, 1935, and Miller, 
1941, and the present experiments. The results 
of these authors may be used as evidence proving 
the existence of a respiratory gradient in the chick 
head-process embryo. There is one objection 
which may be raised to such an interpretation. 
The node region which shows the highest rate of 
Tanus green reduction under the induced anaero- 
IdIc conditions of their experiments is the thickest 



162 



THE COLLECTING NET 



[ Vol. XVI, No. 146 



region of the head-process area pelliicida. Such 
a region ought to show the presence of the re- 
duced form of the dye Ijefore other regions since 
it contains a greater number of cells per unit area 
of pellucida area material. In confirmation of this 
morphological picture of the head-process embryo 
the present observations show that there is the 
greatest amount of nitrogen per unit area in the 
node region. Furthermore, the relative thick- 
nesses and the relative quantities of nitrogenous 
material per unit area agree with the relative rates 
of dye reduction reported. It seems unlikely that 
the dye experiments really show a gradient in the 
respiratory activity of individual cells in the vari- 
ous observed regions. 

It cannot be denied that when it is possible to 
make determinations on even smaller amounts of 
material than were used in the present work some 
significant differences in rate of oxygen consump- 
tion may be found. Furthermore, oxygen con- 
sumption is only one respiratory activity of living 
cells. When other metabolic properties like gly- 



colysis are investigated for the same stages, re- 
gional differences may be discovered. 

By way of conclusion this work is considered a 
phase of descri]5ti\'e embryology. It might be 
called specifically chemical morphology. Com- 
plete analytical investigations based on the present 
data can only come sometime in the future when 
the very general function, the Qoo. can be dis- 
sected into the component factors from which it is 
derived. Only these factors can give some insight 
into the way energy from combusted food sub- 
stances is used in the various embryonic processes 
which result in differentiation. Furthermore, un- 
limited possibilities exist in the manifold consti- 
tution of all the components of the respiratory 
systems for the provision of a variety of metabolic 
differentiations without it being necessary that 
differentiation be reflected in the over-all rates of 
oxygen consumption. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on 
August 5.) 



STUDIES ON CONDITIONS AFFECTING THE SURVIVAL IN VITRO OF A 
MALARIAL PARASITE {PLASMODIUM LOPHURAE) 

Dr. William Tracer 

Rockefeller Institute. Princeton. N. J. 



The malaria parasites comprise a rather homo- 
geneous group of protozoa which inhabit the 
blood and blood forming cells of certain verte- 
brates. They are transmitted from one vertebrate 
host to another by an insect vector in which they 
undergo a se.xual cycle of de^■elo]5ment. There 
are 4 species of malaria parasites of human beings, 
no one of which has e\'er been successfully trans- 
mitted to any other host. There are several species 
of malaria parasites of monkeys and these again 
are highly specific for monkeys. Then there are 
about 12 species of malaria parasites of birds, 
which are specific for certain groups of birds. 
These bird malaria parasites provide good material 
for experimental studies in malaria and one of 
them. Plasuwdiiiui lophurae. has been used for 
the work to l^e reported here. It is an especially 
good organism for experimental purposes as it is 
infectious to the young chicken, a cheap and read- 
ily available host animal. 

All the malaria parasites have an essentially 
similar life cycle. In the vertebrate host, a small 
form, the merozoite. invades a red blood cell and 
grows in size. Its nucleus then divides repeatedly 
to form a segmenter having perhaps 6 to 20 nu- 
clei, depending on the species. A small amount 
of cytoplasm gathers around each nucleus and the 
host red cell bursts and liberates the newly formed 
merozoites. Each merozoite is then capable of 
infecting a new red cell and repeating the process. 
This asexual cycle continues freely in any one 



host until the host succumbs to the infection or 
develops a resistance. Under laboratory condi- 
tions, the asexual stages can be kept going in- 
definitely by the inoculation of blood from an in- 
fected to a fresh host. Since all the experiments 
]Dresented in this paper were concerned exclusive- 
ly with the asexual stages, the sexual stages which 
occur in the mosquito vector need not be con- 
sidered. 

It is evident from this brief review of the biol- 
ogy of malaria parasites that they are highly spe- 
cialized organisms. So far as present knowledge 
goes, they are just as much obligate intracellular 
parasites as are any of the viruses. No one of 
them has ever been cultured in vitro. Indeed, 
previous attempts at their cultivation have failed 
not only of their ultimate object, but have also 
failed to give much information concerning even 
the simplest conditions which might favor the sur- 
vival of the parasites in vitro. 

It therefore seemed desirable to make a com- 
parative study of the effects of various environ- j 
mental conditions on the length of life of the para- 
sites outside of their living host. In these studies 
I have attempted to find, not conditions for the 
preservation of the parasites in a state of sus- 
pended animation, as at very low temperatures, 
but conditions which would favor their survival 
under circumstances which would ]5romote either 
some development or rapid death. Hence, all the 
experiments were performed at temperatures of 



August 23, 1941 ] 



THE COLLECTING NET 



163 



40-42°C., about the body temperature of the 
chicken. Now, how can we judge the survival of 
malaria parasites? They are small and tyijically 
non-motile. Something; can be said as to their 
condition on the basis of their microscopic ap- 
pearance in fresh and stained films, but the only 
reliable criterion of survival is their ability to in- 
fect a susceptible host — in this case a baby chick. 

The experiments were conducted in the follow- 
ing manner. The media to be tested were placed, 
with aseptic precautions, in appropriate sterile 
containers. Blood was taken aseptically from the 
heart of an infected chicken and the red cells were 
centrifuged down and resuspended in a special 
balanced salt solution. Suitable amounts of the 
parasitized red cell suspension were measured into 
the experimental containers. These were held in 
an incubator and removed daily for the taking of 
a small sample. Each sample was used for the 
preparation of a stained smear and for the inocu- 
lation of two 2-day old chicks. If these chicks 
showed an infection by the 7th day after inocula- 
tion, the infectivity of the sample was designated 
+ -f-. If they showed no infection on the 7th day, 
but did show an infection on the 11th day, the in- 
fectivity was designated -\-. If they showed no 
infection by the 11th day, the infectivity was — . 

The balanced salt-glucose solution, called solu- 
tion K, used for the preparation of media and 
parasitized red cell suspensions, was prepared on 
the basis of available knowledge of the inorganic 
composition of red blood cells, and of certain gen- 
eral considerations. It differs from solutions such 
as Locke's and Tyrode's chiefly in having a much 
higher potassium content, a somewhat higher 
phosphate and magnesium content and a lower 
pH (7.2). Comparative tests of survival of P. 
lophitrae in solution K and in Locke's or Tyrode's 
showed always longer survival in solution K. An- 
other factor which was early found to have a fa- 
vorable effect on survival was the presence of red 
cell extract. This was prepared by freezing and 
thawing chicken red blood cells once. The frozen- 
thawed material was suspended in solution K and 
a clear extract obtained by centrifuging out the 
nuclei and cell remnants. Such red cell extracts 
were used in tests of all the other factors to be 
considered. 

Adequate aeration had a marked effect on sur- 
vival. For example, parasites in red cell extract 



in a tulie to a depth of 15 mm. showed no in- 
fectivity by the 3rd day, while those in the same 
red cell extract in a flask to a depth of 4 mm. still 
had a +-(- infectivity on the third day. In a 
preparation held in a vial with air bubbled 
through the infectivity was + on the 5th day, 
while in the control without a current of air the 
infectivity was already — on the 3rd day. But 
if pure oxygen was bubbled through, survival was 
shorter than with COo — free air. 

If fairly dilute red cell extracts were used, an 
effect of the added carbohydrate in the solution K 
could be found. Twelve millimols of glucose per 
liter gave a -)-+ infectivity on the 4th day, as 
compared with a — infectivity for 8 millimols of 
glucose per liter. Again, on the 3rd day, 12 mil- 
limols of glucose per liter gave a + infectivity, no 
added glucose a — infectivity, and 24 millimols 
per liter a — infectivity, showing a toxic effect of 
high glucose concentration. Glucose could be re- 
placed by glycogen. Thus, on the 4th day, the in- 
fectivity with no added carbohydrate was — , with 
0.2% glycogen it was -| — \-. In such dilute red 
cell extracts added glutathione affected survival. 
With 12 millimols of glucose as added carbohy- 
drate, the infectivity was -|- on the 4th day and — 
on the 5th day in the absence of added glutathi- 
one, while in the presence of added glutathione it 
was +4- on the 4th day and still -|-+ on the 5th 
day. 

In a similar way, it has been found that the sur- 
vival in vitro of P. lophurae. as judged by infec- 
tivity, is favored not only by a balanced salt solu- 
tion of high potassium content, by red cell extract, 
by adequate aeration, by appropriate concentra- 
tions of glucose or glycogen and by glutathione, 
but also by daily renewal of the medium, by a 
suitable density of parasites per cu. mm. and by 
certain concentrations of chick embryo extract, 
chicken liver extract and chicken serum or plasma. 

In the best preparations, at least 40% of the 
l^arasites were alive on the 3rd day, at least 20% 
on the 4th day, about 17f on the 5th day and 
about 0.057^ on the 6th day, the last day of the 
tests. In such preparations there was a small in- 
crease in oarasite number on the first day of life 
in z'itro. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on 
August 12.) 



THE EFFECT OF DYES ON RESPONSE TO LIGHT IN PERANEMA TRICHOPHORUM 

Dr. Charles C. H.vssett 



Department of Zoology, Tin 

Peraneina trichophornm is a colorless flagellate 
without an eyespot. It moves by swimming or by 
crawling; when it crawls it moves slowly and is 
Easily kept under observation. It responds to a 
sudden increase in luminous intensity by ceasing 



Johns Hopkins C'niz'ersity 

forward motion, bending sharply and then moving 
off at an angle to its former line of progression. 
This response is known as a shock-reaction. 
Mast and Hawk (1936) and Shettles (1937) 
studied various phases of this response. Shettles 



164 



THE COLLECTING NET 



[ Vol. XVI, No. 146 



showed that cocaine chloride increases the reac- 
tion-time and that strychnine sulfate decreases it. 
In the present work it was decided to study the 
effects of dyes on the response. 

Raab, in 1900, found that the dye acridine pos- 
sessed the power to sensitize paramecia to light 
and that animals in acridine solutions were un- 
harmed if kept in the dark, but were killed if il- 
luminated ; the stronger the light, the more quick- 
ly they were killed. 

The object of these experiments was to ascer- 
tain ( 1 ) the nature of the effect of dyes on the 
reactions of Perancuia and (2) the importance of 
some of the factors involved, e.g., fluorescence, 
wave-length absorbed by the dye, structure of the 
dye molecule. 

Neutral red, eosin, rose bengal, orange G, aura- 
mine O, brilliant green, methylene blue and Nile 
blue sulfate were used; each dye was dissolved in 
Chalkley solution (the culture fluid used in grow- 
ing the peranemae ) , and its absorption spectrum 
was measured on a Coleman spectrophotometer. 
All of the dyes except neutral red and brilliant 
green were found to absorb light in relatively 
limited parts of the visible spectrum ; as a group, 
they cover all parts of the spectrum. 

The peranemae were mounted on slides in 
various concentrations of each dye, then were put 
on the stage of a microscope, brought to focus in 
weak red light and suddenly stimulated by the 
application of a light of 20.35 meter candles in- 
tensity. The reaction-time was measured with a 
stop watch. This was repeated with 50 animals 
at each of a number of concentrations of each dye. 
Two hundred animals from the same cultures 
were used as controls, they were tested while 
mounted on slides in the regular culture fluid. 

The results were as follows : the reaction-time 
of animals which were not treated with dyes was 
found to be 12.1 seconds. Several dyes were 
found to decrease the reaction-lime to approxi- 
mately 1 sec. when the concentration of dye was 
2 X 10"^ M, these were rose bengal, eosin, neu- 
,tral red and methylene blue. More dilute solu- 
tions gave longer reaction-times up to 12 sec. for 
concentrations of 1 X 10*' M. Nile blue sulfate 
and auramine O were less effective, relatively 
strong solutions only produced a minimum reac- 
tion-time of ca. 5 sec. Orange G had no effect. 
Brilliant green increased the reaction-time at a 
concentration of 5 X 10"^ M ; more dilute solu- 



tions produced shorter reaction-times down to 
12 sec. at 1 X 10"^ M. This dye is much more 
to-xic than the others used and the increase in re- 
action-time which was observed may have been 
caused by the toxicity of the dye. 

All the dyes used are lethal to Pcrancma even 
in the dark when strong solutions are used. The 
observations were therefore made with solutions 
which had no harmful effects after as long as 24 
hours in the dark. 

From these results several conclusions can be 
drawn. Concerning fluorescence, in this experi- 
ment the dyes used were of varying degrees of 
fluorescence, the order being approximately : 
eosin > rose bengal, neutral red > Nile blue 
sulfate, methylene blue > auramine O, orange G, 
brilliant green. Comparing this with the order of 
effectiveness in decreasing the reaction-time of 
Perancuia, it can readily be seen that there is little 
correlation. These results agree with those of 
Raab ( 1900), who found that fluoresced radiation 
from dyes not in contact with paramecia had no 
harmful effects. 

The molecular structure of the dyes is diverse, 
(Conn, 1940). Of the four most effective dyes, 
eosin and rose bengal are similar to each other 
but differ greatly from neutral red and methylene 
blue. Hence no specific structural characteristics 
seem to be involved in photodynamic action. 

The wave-length of light necessary to produce 
photodynamic action is dependent on the absorp- 
tion of the dye being used. Although white light 
was used in this work, it has been shown (Blum. 
1941), that action spectra coincide closely with 
absorption spectra. The dyes which were effec- 
tive in reducing the reaction-time of Pcrancma 
vary from auramine O, which absorbs light main- 
Iv in the region 3900-4300 A to methylene blue, 
which alisorbs mostly red light of 5800-6400 A. 

In summary : ( 1 ) several dyes were found to 
exert a photodynamic effect by reducing the reac- 
tion-time of Pcrancma ; ( 2 ) the effect of these 
dyes is inversely proportional to their concentra- 
tion ; (3) no direct correlation was found between 
the effect of the dyes and degree of their fluores- 
cence ; (4) several types of dye molecules are ef- 
fective ; ( 5 ) the active dyes possess diverse ab- 
sorption spectra. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on 
August 12.) 



PROGRAM OF THE SUMMER MEETINGS OF THE GENETICS SOCIETY OF AMERICA 
AT COLD SPRING HARBOR, LONG ISLAND, N. Y., AUGUST 27 TO 29, 1941 



Officers of the Genetics Society of America 

President, Th. Doezhanskv, Columbia University. 
Vice-President, E. \V. Lindsteom, Iowa State College. 
Secretary and Treasurer, B. P. Kaupman, Carnegie In- 
stitute of Washington. 



Wednesday, August 27, 8:30 P. M., Blackford Hall, 
Evening Lecture 

A. 11. Sturtevant, California Institute of Technology* 
Comparative Genetics of the Species of Drosophila. 



August 23, 1941 ] 



THE COLLECTING NET 



Thursday, August 28, 9:30 to 12:45, Davenport 
Laboratory, Demonstrations 

(1) A'rwooD, Sanford, S., U. S. Regional P.Tsturo 
Rt'scarcli Laboratory, State College, Pa.: The multiple 
oi)positional alleles causing cross-incompatibility in Tn- 
folvum re pens. 

(2) Bishop, D. W., University of Pennsylvania, 
Philadelphia, Pa. : Cytologieal demonstrations of chro- 
mosome breaks soon after X-radiation. 

(3) Brehme, Katherinb S., Carnegie Institution of 
Washington, Cold Spring Harbor, N. Y. : A survey of 
the Malpighian tube color of the eye color mutants of 
DrosophiJa me!anogaster. 

(4) BusHNELL, Ealph J., The University of Con- 
necticut, Storrs, Conu. : Incompatible matings in inbred 
families of the bean weevil. 

(5) Demerec. M., Hollaendek, Alexander, Houla- 
HAN, M. B., and Bishop, M., Carnegie Institution of 
Washington, Cold Spring Harbor, N. Y., and National 
Institute of Health, Bethesda, Md. : Effect of mono- 
chromatic ultraviolet radiation on Drosophila melano- 
gaster. 

(6) EiGSTi, O. J., University of Oklahoma, Norman, 
Ckla.: A comparative study of the effects of sulfanila- 
mide and colchicine upon mitosis of the generative cell 
ill the pollen tube of Tradescantia oecidentalis (Brit- 
ton) Smyth. 

(7) Pankhauser, Gerhard, and Crotta, Eita, 
Princeton University, Princeton, N. J. : The frequency 
of spontaneous aberrations of chromosome number 
among larvae of the newt, Triturus viridescens. 

(8) Gordon, Miron, New York Aquarium, New 
York, N. Y. : Dominant and recessive responses of the 
Sd factor in natural and domesticated fish populations. 

(9) HiNTON, O. Taylor, and Atwood, K. C, Colum- 
bia University, New York, N. Y. : A comparison of the 
specificities of terminal adhesions of salivary gland 
chromosomes in two strains of Drosophila. 

(10) Kamenopf, Ralph J., City College, New York, 
N. Y. : A cytologieal study of the embryonic livers (16- 
18 days) of normal and flexed-tailed (anemic) mice. ' 

(11) Kimball, R. F., Johns Hopkins University, 
Baltimore, Md. : A gene affecting the manner of swim- 
ming in the ciliate protozoan, Euplotes patella. 

(12) Laanes, T., and MacDowell, E. C, Carnegie 
Institution of Washington, Cold Spring Harbor, N. Y. : 
Screw-tail, a new mouse mutation. 

(13) Power, Maxwell E., Yale Universitty, New 
Haven, Conn. : Neurological effects of mutants reducing 
facet number in the eyes of Drosophila melanogaster. 

(14) Riddle, Oscar, Dunham, H. H., and Schooley, 
J. P., Department of Genetics, Carnegie Institution of 
Washington, Cold Spring Harbor, N. Y. : Genetic her- 
maphroditism in a strain of pigeons. 

(15) Robertson, G. G., Yale University, New Haven, 
Conn. : Increased viability of homozygous yellow mouse 
embryos in new uterine environments. 

(16) SONNENBLicK, B. P., Queens College, Flushing, 
N. Y. : The question of cell constancy in various em- 
bryonic and larval tissues of Drosophila melanogaster. 

(17) Sparrow, A. H., McGill University, Montreal, 
Canada : Spiralization in microspore chromosomes of 
Trillium. ^ 

(18) Spencer, W. P., College of Wooster, Wooster, 
Ohio : Inherited variations in wild populations of Clay- 
tonia virginica. 

(19) Wilson, G. B., and Boothboyd, E. R., McGill 
University, Montreal, Canada: Differential reactivity in 
the chromosomes of Trillium species. 

(20) Wilson, G. B., and Sparrow, A. H., McGill 
University, Montreal, Canada: Partial fusion of un- 
treated root tip chromosomes of Trillium erectum L. 

Afternoon Session, beginning at 2 o'clocit 

Inspection of Exhibits prepared by resident investiga- 
tors of the Department of Genetics, Carnegie Institu- 



165 

tioii of Washington and of the Biological Labor.-itory 
of flic Long island Biological Association. 
Late Afternoon and Evening 

Swim, I'unic Sapper, and Dance. 

Friday, August 29, at 9:30 A. M., Auditorium of 
Blackford Hall 

(1) Bernstein, Marianne E., Carnegie Institution 
of Washington, Genetics Record Office, Cold Spring 
Harbor, N. Y. : The incidence and Meudelian transmis- 
sion of mid-digital hair in man. 

(2) Lindegren, Carl C, and Lindegren, Gertrude, 
University of Southern California, Los Angeles, Calif.: 
X-ray and ultraviolet induced mutations in Neurospora. 

(3) Brink, R. A., and Cooper, D. C, University of 
Wisconsin, Madison, Wis.: Somatoplastic sterility as a 
function of the endosperm genotj'pe. 

(4) Nebel, B. R., Wilson, G. B., and Maeinelli, L., 
Ne\v York Agricultural Experiment Station, Geneva, N. 
Y., McGill University, Montreal, Canada, and Memorial 
Hospital, New York, N. Y.: X-ray dosage curves in 
Tradescantia. 

(5) Giles, N. H., and Nebel, B. R., Yale Univer- 
sity, New Haven, Conn., and New York Agricultural Ex- 
periment Station, Geneva, N. Y. : An analysis of the 
intensity factor in X-ray induced chromosomal aberra- 
tions in Tradescantia. 

(6) Caspaei, Ernst, Lafayette College, Easton, Pa.: 
Genetic and environmental factors influencing testis color 
in Ephest.a kuhniella. 

(7) Muller, B. H., and Pontecorvo, G., Amherst 
College, Amherst, Mass., and the University of Edin- 
burgh, Edinburgh, Scotland: Recessive genes causing 
interspecific sterility and other disharmonies between 
Drosophila melanogaster and simulans. 

(8) Lewis, E. B., California Institute of Technol- 
ogy. Pasadena, Calif.: The Star and asteroid loci in 
Drosnphi'a melanogaster. 

(9) Steinberg, Arthur G., McGill University, Mon- 
treal, Canada: Further studies on the histological devel- 
opment of the wild type and Bar eyes of Drosophila 
melanogaster. 

(10) Ives. P. T., Amherst College, Amherst, Mass.: 
Allelism and elimination of lethals in American popula- 
tions of Drosophila melanogaster. 

(11) Neel, J. v., Dartmouth College, Hanover, N. 
H. : A case of high mutation frequency in DrosoplMa 
melanogaster. 

Afternoon Session 

Inspection of Exhibits — Further opportunity to visit 
exhibits prepared by resident investigators. 



THE GROWTH OF THE LABORATORY 

The following table shows the total number of 
investig-ators registered at the Marine Biological 

Laboratory each year since its foundation. 

1888 8 1906 68 1924 194 

1889 19 1907 60 1925 207 

1890 20 1908 52 1926 252 

1891 24 1909 66 1927 294 

1892 50 1910 62 1928 323 

1893 41 1911 82 1929 329 

1894 58 1912 93 1930 337 

1895 53 1913 122 1931 362 

1896 74 1914 129 1932 314 

1897 58 1915 137 1933 319 

1898 70 1916 129 1934 323 

1899 71 1917 129 1935 315 

1900 70 1918 93 1936 359 

1901 65 1919 134 1937 391 

1902 76 1920 136 1938 380 

1903 76 1921 172 1939 352 

1904 51 1922 182 1940 386 

1905 68 1923 176 



166 



THE COLLECTING NET 



[ Vol. XVI, No. 146 



The Collecting Net 

A weekly publication devoted to the scientific work 
at marine biological laboratories. 

Edited by Ware Cattell with the assistance of 
Boris I. Gorokhoff and Judy Woodring. 

Entered as second-class matter, July 11, 1935, at 
the U. S. Post Office at Woods Hole, Massachusetts, 
under the Act of March 3, 1879, and re-entered, 
July 23, 1938. 

SUMMER ACTIVITIES OF UNIVERSITY OF 
CHICAGO BIOLOGISTS 

Dr. Carl Moore, professor and chairman of the 
department of zoology, is spending the summer 
in the north woods at Rapid City, Michigan, 
working in his own private laboratory on the 
hormone control of reproduction. In January he 
received the American Academy of Arts and 
Sciences award for his work which served as a 
basis for the isolation and identification of "tes- 
tosterone", the male sex hormone. 

Dr. Sewall Wright, Ernest D. Burton Distin- 
guished Service professor of zoology, is at Cold 
Spring Harbor. Long Island. 

Dr. Alfred Emerson, professor of zoology, is 
conducting an inspection trip of eastern museums, 
using his summer home at Hewlett Landing in 
upper New York as a base. He is a specialist in 
the field of ecology. 

Dr. Frances Oldham, research assistant in the 
department of pharmacology, is spending her sec- 
ond summer at the Eureka Whaling station at 
Field's Landing in Northern California. At this 
station the whales are carved and prepared in 
public — for an admission charge — before they are 
marketed. Dr. Oldham will bring back pituitary 
glands for her work with Dr. Eugene Ceiling, 
professor and chairman of the department of 
pharmacology. 

M.B.L. TENNIS CLUB 

Two of the five tennis tournaments this year 
have already been completed : the finals in the 
other three are scheduled for this afternoon. 

In the finals of the men"s singles. D. E. Lance- 
field defeated A. Sttmkard, 6-6, 7-5. 

In the finals of mixed doubles. Lancefield and Te 
Winkel defeated Lumb and Hunim, 6-2. 6-2. 

The ladies' singles is one of the tournaments 
scheduled to be completed this afternoon. Mary 
Chamberlain will be one of the finalists, having 
defeated P. Saunders, 7-5. 6-3, and she will play 
against D. Baitsell, who had previously defeatel 
G. Gorokhofif, 6-2, 6-4. 

The finals of the men's doubles will see Stunk- 
ard and Evans ( who have defeated Jones and 
Speidel, 6-2, 6-1 ) playing against the winners of 
the semi-finals in the other bracket — Humm and 
Hayashi vs. Lancefield and Krahl. 

The finals of the junior singles are also sched- 
uled for this afternoon. 



WOODS HOLE CHORAL SOCIETY 

The fourteenth annual concert of the Woods 
Hole Choral Society will be presented Monday 
evening at 8 :30 in the Woods Hole Community 
Hall. Twelve numbers, equally divided between 
secular and sacred music, have been prepared by 
the members of the Club, who have been rehears- 
ing twice weekly since the beginning of the sum- 
mer. 

The concert will be conducted by Professor 
Ivan T. Gorokhoff, director of choral music at 
Smith College. Dr. Eliot R. Clark is president 
of the Club, and Dr. Charles Packard is secretary- 
treasurer. Miss Galina Gorokhofif is accompanist. 

The members of the Society, which is composed 
primarily of Laboratory workers and members of 
their families, are as follows : 

Stella Anderson, Barbara Brainerd, Jane Collins, 
Grace Crecelius, Emily H. Lower, Edith Mitchell, 
Louise Thorne Sprenger, Evelyn G. Watterson, 
Helen Willier, Eleanor Linton Clark, Helen M. 
Crossley, Eva Stokey Evans, Alma G. Stokey, Wil- 
liam J. Blake, Stanley Sprenger, Peter J. Wilhousky, 
William H. Batchelor, John W. Brainerd, Eliot R. 
Clark, Boris I. Gorokhoff, Arthur Truslow, and 
George G. Lower. 

The program is as follows : 
Break forth, O beauteous, heav'nly light J. S. Bach 
Hail Holy Light! Alexander Kastalsky 

Rejoice in the Lord alway Henry Purcell 

Ave Verum Corpus William Byrd 

Gospodi Pomiluy (Lord, Have Mercy) Lvovsky 

When His loud voice (Jeptha) Georg F. Handel 

Brightly Dawns Our Wedding Day (The Mikado) 

Gilbert and Sullivan 
Good-day, dear heart Orlando di Lasso 

The Beetle's Wedding German Folk-Song 

A Legend P. Tschaikowsky 

Round the Good Father's Door A. Arkhangelsky 
Moon Magic Three Russian Folk Songs 

M.B.L. CLUB 

The jiing pong tournament of the M.B.L. Club 
is well under way with most of the first and some 
second rounds played ofif'. The tournaments or- 
ganized this year are men's and women's singles 
and mixed doubles. 

The Monday evening phonograph record con- 
cert will not be given this week in order to avoid 
a conflict with the Choral Club concert. 



CURRENTS IN THE HOLE 

At the following hours (Daylight Saving 
Time) the current in the Hole turns to run 
from Buzzards Bay to Vineyard Sound: 
Date A. M. P. M. 

August 24 5:30 5:50 

Atigust 25 6:15 6:37 

August 26 7:01 7:27 

August 27 7:49 8:19 

August 28 8:41 9:16 

August 29 9:35 10:15 

Augttst 30 10:35 11:19 



August 23, 1941 ] 



THE COLLECTING NET 



167 



ITEMS OF INTEREST 



Dr. Dorothy M. Wrinch was married on 
W'ednesdav at 12:30 P. M. in the Lalioratory to 
Dr. Otto Glaser by Rev. Ralph H. Lon^ of Fal- 
mouth. They are spending their honeymoon on 
Nantucket Island, and will return later to their 
house in Ouisset. Dr. Wrinch will continue her 
scientific work as liefore ; she holds a professor- 
ship in molecular biology in Dr. Glaser's depart- 
ment of biology at Amherst, and is also professor 
at Smith College and Mt. Holyoke College. She 
was given in marriage by Professor John F. Ful- 
ton of the Yale University School of Medicine. 
Dr. Katherine Brehme was her only attendant. 
The marriage is the fitst to have been performed 
in the Laboratory. 

Dr. Henry J. Fry died on August 15 in 
Hartford, Connecticut, after a long illness. Dr. 
Fry first came to Woods Hole as a student in 
1921 and returned as an investigator nearly every 
year thereafter until 1939. He received his A.B. 
degree at Muhlenberg College in 1914, and his 
Ph.D. at Columbia University in 1925. In 1923 
he began instructing at New York University, 
becoming a professor in 1930. From 1933 until 
his illness he was a visiting investigator at Cor- 
nell University Medical College. Dr. Fry was 
noted for his work in experimental cytology. 

New members of the M.B.L. Corporation in- 
clude: R. Ballentine, M. M. Brooks, Aurin M. 
Chase, K. W. Cooper, Titus C. Evans, Jean M. 
Fisher, C. G. Grand, J- Friedrich Gudernatsch. 
Rudolf T. Kempton, Otto Loewi, F. M. Mac- 
Naught. V. J^Ienkin. Isabel M. Morgan and K. G. 
Stern. 

Mr. David D. Perkins, who graduated from 
the University of Rochester this spring, and who 
is taking the invertebrate zoology course at the 
Marine Biological Laboratory, has been appointed 
a graduate assistant in zoology at Columbia Uni- 
versity. 

Mr. R. a. Goffin, superintendent of the Bu- 
reau of Fisheries, reports that 11,626 visitors to 
the aquarium registered in the guest book between 
July 13 and August 16. Since only about one 
quarter of the visitors register, the actual numljer 
probably was about four times as great. 

The Anton Dohrii left the Oceanographic In- 
stitution Wednesday for Nova Scotia to conduct 
research work in connection with national defense. 

At the annual meeting of the Tennis Club, the 
following persons were elected officers : President, 
Eric Ball ; Vice-President, Margaret Speidel : 
Secretary-Treasurer. Titus Evans. D. E. Lance- 
field and E. R. Jones were elected to the execu- 
tive committee. 



Professor A. H. Sturtevant will give the 
evening lecture at the summer meetings of the 
Genetics Society of America at Cold Spring Flar- 
l(or on August 27. He will speak on "Compara- 
tive Genetics of the Species of Drosophila." 

Mr. Robert L. Terry, who was an investiga- 
tor from the University of Pennsylvania at the 
Laboratory last year is now a private in the U. 
S. Army. 

Dr. T. H. Johnson, assistant director of the 
Bartol Research Foundation, has been working at 
the Oceanographic Institution for brief periods 
during the summer. 

Dr. J. H. McGregor of Columbia University, 
left Woods Hole on Thursday after a two-day 
visit. 

Dr. Harlan T. Stetson, director of cosmic 
terrestrial research at the Massachusetts Institute 
of Technology, recently visited Woods Hole on 
his vessel, the Calypso. 

Dr. and Mrs. M. W. Bosworth. both of 
whom were at the Laboratory in 1940, visited 
Woods Hole briefly last week-end. 

Among others visiting the Laboratory recently 
have been Drs. William F. Diller, Irene Corey 
Diller, H. K. Hartline. I. M. Korr, Felix Bern- 
stein, Ernst Fischer, and D. Eugene Copeland. 

Dr. Robert Chambers will leave Monday for 
New York to attend a conference on tissue cul- 
ture. 

Mr. Carl Alper, a student janitor at the Lab- 
oratory, injured his foot in an elevator and re- 
turned' to his home in New Jersey last Sunday. 

Dr. Kenneth C. Fisher has left for St. Johns, 
New Brunswick, to visit his father, who is ill. 

Professor and Mrs. L. L. Woodruff last 
Sunday afternoon gave a tea at their home in 
Gansett for the present and former members of 
the Osborn Zoological Laboratory, Yale Univer- 
sity, who are at Woods Hole this summer. About 
thirty were present. 

Mr. Edward Chambers has received a Rocke- 
feller Foundation grant to conduct research work 
at the Columbia County Department of Public 
Health in Hudson, New York. He and Mrs. 
Chambers will leave Woods Hole on Monday. 
Mr. and Mrs. Chambers entertained at Edgewood 
Monday afternoon for about seventy-five meml^ers 
of the Laboratory and their families with a garden 
narty at which a dramatization of a Russian fairy 
tale was presented. Dr. Robert Chambers acted 
as narrator throughout the play. 



168 



THE COLLECTING NET 



[ Vol. XVI, No. 146 



INVERTEBRATE CLASS NOTES 



Dr. Bissonnette's one-day survey of the Bryo- 
zoa was for many an introduction to this group. 
The lab study of the living material fixed in our 
minds at least some of the salient features of this 
group. 

Though the Molluscs are no novelty. Dr. Mat- 
tox had no trouljle keeping us interested in his 
discussions. Of particular interest in the lab 
work were the dissections of Busycoii and the 
squid, and the study of the Pelicypod heart in its 
reactions to salt solutions and drugs. 

Ann Weber's organization and the splendid 
weather proved an unbeatable combination for the 
picnic at Tarpaulin Cove. Some ambitious stud- 
ents made plans for competitive collecting team.-^, 
but every one was glad to aljandon the project in 
favor of baseball. Several small groups spent the 
morning collecting or watching the birds. Seventy 
of us students, instructors and families made a 
full load for the A^ercis and the Winifred. The 



remnant of forenoon was just sufficient time to 
build up an appetite for the picnic lunch of sand- 
wiches and clams. Old friends of the lab aquaria, 
Mytilus and Mya, made an abundant feast. Mrs. 
Rankin, Randy Kielich and Bob Williams en- 
gaged in a vvatermelon-seed-blowing contest, in 
which Mrs. Rankin was the victor. 

After lunch a game of M-ball was begun. To 
quote from the sports-columnist for the "Inverts," 
Johnny Osmun : 

"Soon to be released through national sports 
writers are the rules for M-ball. Originating at 
and named for the M.B.L., this game is played 
literally and figuratively with the head. Nearly 
every day the varsity and occasionally the fresh- 
men (faculty, principally) may be seen at Kahler 
stadium bouncing a ball from head to head. High 
score of nineteen uninterrupted bounces." 

— Louise Gross and Bill Batchclor 



SOME RECENT BOOKS IN THE BIOLOGICAL SCIENCES 



Ackerman, E. A. New England's Fishing Industry. 
$4.00. Chicago. 

Adams, N. E., and E. M. Bandow. A Study Guide 
for AppHed Biology. $1.75. Burgess. 

Alexander, G. An Outline of General Zoology. 
$1.00. Barnes and Noble. 

Allen, P. W., and G. M. Cameron. Microbiology 
Laboratory Manual. $1.50. Mosby. 

Arey, L. B. Developmental Anatomy (4th ed.). 
$6.75. Saunders. 

Arnold, J. G., and T. L. Duggan. Laboratory Man- 
ual of General Biology (4th ed.). $1.50. Mosby. 

Baitsell. G. A. Human Biology. $3.75. McGraw- 
Hill. 

Baitsell, G. A. Manual of Biology (6th ed.). $2.75. 
Macmillan. 

Baly, E. C. C. Photosynthesis. $4.75. Van Nostrand. 

Beaver, W. C. Fundamentals of Biology (2nd ed.). 
$4.00. Mosby. 

Baaver, W. C. Laboratory Outlines of General Biol- 
ogy (2nd ed.). $2.00. Mosby. 

Bell, D. J. Introduction to Carbohydrate Biochem- 
istry. University Tutorial (London). 

Biological Laboratory, Cold Spring Harbor. Sym- 
posia on Quantitative Biology. Vol. VIII. Darwin 
Press. 

Biological Symposia. Vol. II, §2.50. Vol. Ill, $3.50. 
Jaques Cattell Press. 

Blum, H. F. Photodynamic Action and Diseases 
Caused by Light. $6.00. Reinhold. 

Brockleshy. H. N.. ed. Bulletin No. LIX: The Chem- 
istry and Technology of Marine Animal Oils 
with Particular Reference to Those of Canada. 
$2.95. Fisheries Research Board of Canada. 

Burnet, F. M. Biological Aspects of Infectious Di- 
sease (4th ed.). $3.75. Cambridge (Macmillan). 

Cable, R. M. An Illustrated Laboratory Manual of 
Parasitology. $1.50. Burgess. 

Calkins, G. N., and F. M. Summers, ed. Protozoa in 
Biological Research. $10.00. Columbia. 

Cameron, A. T. Recent Advances in Endocrinology 
(4th ed.). Churchill Ltd. 

Cameron, G. M. Bacteriology of Public Health. 
S3. 50. Mosby. 

Carlron, A. J., and V. Johnson. The Machinery of 
the Body. $4.00. Chicago. 



Carroll, P. L., and W. P. Horner. Atlas of the Frog. 
S1.25. Mosby. 

Carter, G. S. A General Zoology of the Inverte- 
brates. $5.50. Macmillan. 

Chandler, A. C. — Introduction to Parasitology; with 
Special Reference to the Parasites of Man. 
$5.00. Wiley. , 

Chapman, V. J. Introduction to the Study of Algae. 
$4.00. Cambridge (Macmillan). 

Child, C. M. Patterns and Problems of Develop- 
ment. $8.00. Chicago. 

Clark, C. C. and R. H. Hall. This Living World. 
$3.25. McGraw-Hill. 

Clausen, J., D. D. Keck and W. M. Hiesey. Experi- 
mental Studies on the Nature of Species. $3.50. 
Carnegie. 

Cole, E. C. Text of Comparative Histology. $4.00. 
Blakiston. 

Colin, E. C. Elements of Genetics. S3.00. Blakiston. 

Ccnn, H. J. Biological Stains; A Handbook on the 
Nature and Uses of the Dyes Employed in the 
Biological Laboratory (4th ed.). $3.40. Biotech 
Publications. 

Cross, J. C. An Introduction to Biology. $1.90. Mos- 
by. 

Dahlberg, G. Statistical Methods for Medical and 
Biological Students. $2.75. Interscience. 

Dobzhansky, T. Genetics and the Origin of Species 
(rev. ed.). $4.25. Columbia. 

Eddy, S., C. P. Oliver and J. P. Turner. Atlas of 
Outline Drawings of the Dogfish Shark, the 
Necturus, and the Cat for Vertebrate Anatomy. 
$1.50. Wiley. 

Elvehjem, C. A., and P. W. Wilson. Respiratory 
Enzymes. $3.25. Burgess. 

Evans, H. M. Brain and Body of Fish. $3.50. Blak- 
iston. 

Fasten, N. Introduction to General Zoology. S3. 75. 
Ginn. 

Felt, E. P. Plant Galls and Gall Makers. $4.00. 
Comstock. 

T^ederal Writers' Project. Oysters. $0.50. Whitman. 

Forbes, J. A Laboratory Manual for Histology. 
$1.25. Fordham. 

Frazer, J. E. Manual of Embryology (2nd ed.). 
$9.00. Williams and Wilkins. 



August 2i, 1941 ] 



THE COLLECTING NET 



169 



Frobisher, M., Jr. Fundamentals of Bacteriology 
(2nd ed.). .$2.00. Saunders. 

Fuller, H. J. Outline of General Botany. $0.75. 
Barnes and Noble. 

Fuller, H. J. The Plant World; A Text in College 
Botany. $3.25. Holt. 

Cause, G. F. Optical Activity and Living Matter. 
$2.75. Biodynamica. 

Gerard. The Body Functions. $2.25. Wiley. 

Gerard, R. W. Unresting Cells. $3.75. Harper. 

Glass, H. B. Genes and the Man. Teachers' College, 
Columbia University. 

Graves, J. E., and E. O. Elementary Bacteriology 
(4th ed.). $3.50. Saunders. 

Green, D. E. Mechanisms of Biological Oxidations. 
$2.75. Cambridge (Macmillan). 

Grimes, C. W. Story Outline of Evolution. $2.00. 
Bruce Humphries. 

Gudger, E. W., ed. Archaic Fishes. Breeding Habits, 
Reproductive Organs and External Embryonic 
Development of Chlamydoselachus. American 
Museum of Natural History. 

Guyer, M. F. Animal Biology (3rd ed.). $3.75. 
Harper. 

Hannum, C. A. Comparative Embryology of Verte- 
brates. $4.50. Macmillan. 

Harrow, B. Laboratory Manual of Biochemistry. 
§1.50. Saunders. 

Hewer, H. R. Practical Zoology. $2.00. Chemical 
Publishing Company. 

Hewitt, R. Bird Malaria. $1.10. Johns Hopkins. 

Hill, J. Germs and the Man. $3.50. Putnam's. 

Hogben, L. Principles of Animal Biology. $3.75. 
Norton. 

Holman, R. M., and W. W. Robbing. Elements of 
Botany (3rd ed.). $2.75. Wiley. 

Hoskins, R. G. Endocrinology; the Glands and 
Their Functions. $4.00. Norton. 

Hubbs, C. L., and K. Lagler. Synopsis of the Fishes 
of the Great Lakes Basin. $0.50. Cranbrook. 

Huettner, A. F. Fundamentals of Comparative Em- 
bryology of the Vertebrates. S4.50. Macmillan. 

Huff, G. C. Laboratory Guide for General Biology. 
Swift. 

Hylander, C. J., and O. B. Stanley. Plants and Man. 
$3.00. Blakiston. 

Jennings, H. S. The Beginnings of Social Behavior 
in Unicellular Organisms. $0.50. Pennsylvania. 

Johannsen, O. A., and F. H. Butt. Embryology of 
Insects and Myriapods. $5.00. McGraw-Hill. 

Johlin, J. M. Introduction to Physical Biochemistry. 
$2.75. Hoeber. 

Johnson, P. L. Projects in General Zoology. $1.50. 
Swift. 

Jordan, E. O., and W. Burroughs. Textbook of Bac- 
teriology. Saunders. 

Kendall, J. I. The Microscopic Anatomy of Verte- 
brates. $3.75. Lea and Febiger. 

Kleiner, I. S., and L. B. Dotti. Laboratory Instruc- 
tions in Biochemistry. $1.50. Mosby. 

Krogh, A. The Comparative Physiology of Respira- 
tory Mechanisms. $3.00. Pennsylvania. 

Landis, Carney, et al. Sex in Development. $3.75. 
Hoeber. 

Large, E. C. The Advance of the Fungi. $4.00. Holt. 

Leach, J. G. Insect Transmission of Plant Diseases. 
$6.00. McGraw-Hill. 

Leonard, A. B. Laboratory Guide to Study of De- 
velopmental Anatomy. $1.50. Burgess. 
j Ley, W. The Lungfish and the Unicorn: an Excur- 
j sion into Romantic Zoology. $2.75. Modern Age 

1 Books. 

, Longley, M. H. Laboratory Manual in Bacteriology. 
$1.50. F. A. Davis. 



Luck, J. M., ed. Annual Review of Physiology, Vol. 
Ill, 1941. $5.00. American Physiol. Soc. and An- 
nual Reviews. 

Luyet, B. J., and P. M. Gehenio. Life and Death at 
Low Temperatures. Biodynamica. 

Mavor, J. W. General Biology (rev. ed.). |4.00. 
Macmillan. 

Metcalf, M. M. Further Studies on the Opalinid 
Ciliate Infusorians and Their Hosts. U. S. Na- 
tional Museum. 

Miller, A. H. Speciation in the Avian Genus Junco. 
$3.00. California. 

Miller, E. S. Quantitative Biological Spectroscopy. 
$3.50. Burgess. 

Naturalists' Directory (32nd ed.). $3.00. Cassino 
Press, Salem, Mass. 

Needham, J. Biochemistry of Morphogenesis. $2.00. 
Cambridge (Macmillan). 

Newby, W. W. The Embryology of the Echuiroid 
Worm Urechis caupo. $2.00. American Philo- 
sophical Society. 

Nord, F. F., and C. H. Advances in Enzymology 
and Related Subjects. $5.50. Interscience. 

O'Hanlon, M. E. Fundamentals of Plant Science. 
$4.25. Crofts. 

Olson, O. S. Methods of Teaching High School Biol- 
ogy. 81.25. Burgess. 

Osborn, F. Preface to Eugenics. $2.75. Harper. 

Parker, T. J. A Text-book of Zoology (6th ed.). 2 
vols. $16.00. Macmillan. 

Parshley, H. M. Biology. $2.25. Wiley. 

Perry, J. Fish Production. $1.50. Longmans. 

Perry, L. M. Marine Shells of the Southwest Coast 
of Florida. $4.50. Paleontological Research In- 
stitute, Ithaca. 

Pfeiffer, H. H. Experimentelle Cytologie. Chronica 
Botanica. 

Potter, G. E. Essentials of Zoology. $3.75. Mosby. 

Prescott, S. C, and C. G. Dunn. Industrial Micro- 
biology. $5.00. McGraw-Hill. 

Rabinowitch, E. Photosynthesis. Interscience. 

Ritchie, J. W. Biology and Human Affairs. $2.32. 
World Book. 

Salter, W. T. The Endocrine Function of Iodine. 
$3.50. Harvard. 

Sass, J. E. Elements of Botanical Microtechnique. 
$2.50. McGraw-Hill. 

Schwartz, J. Adventures in Biology. $0.50. Ameri- 
can Biology Teachers. 

Sherrington, C. Man on His Nature. $3.75. Cam- 
bridge (Macmillan). 

Shull, A. F. Principles of Animal Biology (5th ed. 
rev.). $3.50. McGraw-Hill. 

Smith, E. T., and L. M. Weber. Guide to Modem 
Biology. .$1.00. Harcourt. 

Smith, K. The Virus, Life's Enemy. $2.00. Cam- 
bridge (Macmillan). 

Snell, G. D. Biology of the Laboratory Mouse. 
S7.00. Blakiston. 

Stanford, E. E. Man and the Living World. $3.50. 
Macmillan. 

Stiles, K. A. Handbook of Microscopic Characteris- 
tics of Tissues and Organs. $1.50. Blakiston. 

Stiles, K. A. Laboratory Explorations in Animal 
Biology. $2.50. Burgess. 

Stockard, C. R. The Genetic and Endocrine Basis 
for Differences in Form and Behavior as Eluci- 
dated by Studies of Contrasted Pure-Line Dog 
Breeds and Their Hybrids. $7.50. Wistar. 

Svedberg, T., and others. The Ultracentrifuge. 
$11.50. Clarendon. 

Swingle, D. B. General Bacteriology. $3.00. Van 
Nostrand. 

Symposium on Hydrobiology. $3.50. Wisconsin. 

Thone, F. The Microscopic World. $3.00. Messner. 



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Thorsteinson, E. D. New or Noteworthy Amphi- 

pods from the North Pacific Coast. Washington. 
Transeau, E. N., H. C. Sampson and L. H. Tififany. 

Text-book of Botany. $4.00. Harper. 
University of Pennsylvania. Cytology, Genetics and 

Evolution. $2.00. Pennsylvania. 
Visual Aids Service. Visual Aids in the Realm of 

Biology. $0.50. N. J. State Teachers' College. 
WaJdington, C. H. Organisers and Genes. $3.00. 

Cambi'idge (Macmillan). 
Wheeler, W. F. Immediate Biology. $6.00. Chemical 

Publishing Company. 
White, E. G. Principles of Genetics. $2.50. Mosby. 
Willis, J. C. The Course of Evolution by Differen- 



tiation or Divergent Mutation Rather Than by 
Selection. $3.00. Cambridge (Macmillan). 

Wilson, P. W. The Biochemistry of Symbiotic Ni- 
trogen Fixation. $3.50. Wisconsin. 

Windle, W. F. Physiology of the Fetus. $4.50. 
Saunders. 

Wodsedalek, J. E. General Zoology Laboratory 
Guide. $2.90. Burgess. 

Wolcott, R. H. Animal Biology (2nd ed.). $3.50. 
McGraw-Hill. 

Woodruff, L. L., ed. Development of the Sciences, 
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Woodruff, L. L. Foundations of Biology (6th ed. 
rev.). $3.75. Macmillan. 



THE UTILIZATION OF AMMONIA BY CHILOMONAS PARAMECIUM 

(Continued from page 157) 



The material to be presented here is part of a 
comprehensive study of the carbon and nitrogen 
metabolism of Chilomonas. begun two years ago 
at the Harvard Medical School under Prof. A. B, 
Hastings and continued later at the Johns Hop- 
kins University under Prof. S. O. Mast. 

The problem of the source of energy for growth 
of Chilomonas was attacked by Burrows (Proto- 
plasnia. Bd. 31, S. 20-26, 1938), who used mass 
cultures of chilomonads grown in a solution like 
that shown in Table I (Solution A) and in a 
soltition with the acetate omitted liut with a gas 
phase containing a partial pressure of COi; equi- 
valent to 12 cm. of mercury. Growth in both 
solutions amounted to only 5-6 thousand chilomo- 
nads/cc. and the conversion of substrates was too 
small to afford reliable analytical data. 

Unfortunately this sittiation still obtains with 
respect to the inorganic solution, but as shown by 
Htitchens (J. Cell, and Comp. Physiol.. V. 16, pp. 
265-267, 1940) (and unpulilished results) by use 
of a solution to which thiamin and iron are added 
populations up to 1 million cells/cc. can be ob- 
tained (Talile I, Solution B). In addition to the 
Bi and iron it will be noted that the buffering 
capacity of the solution has been increased. In 
addition to this it is necessary to add acetic acid 
from time to time to neutralize the excess base 

Table I. 

Composition oj Solutions Used for Groz^'ing 

Chilomonas 



Substances 




Solution 


mg. 
A 


Solution B 


CHsCOONa 


•3H,0 


249 






249 


NPI4CI 




46 






46 


(NH4).S04 




10 






10 


K,HP04 




20 






150 


MgCU 




1 






1 


CaCl. 




1 






1 


FeCls 




— 






0.17 


Thiamin hyd 


rochloride 


— 






0.01 



left by removal of acetate and to afford additional 
carbon supply. 

In contrast with the old solution in which an 
inoculum of about 500 cells/cc. resulted in a final 
population of 5-6 thousand cells/cc. or a ten-fold 
increase in cell numbers, in this solution an ino- 
culum of 500-1000 cells/cc. results in a final pop- 
ulation of as much as 1 million cells per cc. or an 
increase in cell numbers of 1000 times. Thus 
large numbers of cells, milligrams or even grams 
of material are available and the amounts of sub- 
strates used are readily measurable. 

In experiments which I shall not have space to 
present it has been found that the chilomonads 
utilize large amounts of acetic or lactic acids, ap- 
parently sufficient to account for all of the oxygen 
consumed and to furnish energy for the growth 
obtained in media containing these substances. 
The present experiments deal with the heretofore 
unanswered question of the possible olitaining of 
additional energy by oxidation of ammonia. 

The experimental methods tised are as follows : 
The organisms were grown in sterile, pure, mass 
cultures in 125 cc. Erlenmeyer flasks containing 
50 cc. of the solution shown in Table I (Solution 
B). Inocula of 1-2 thousand organisms/cc. were 
tised, transfers being made from cultures 48 hours 
old. Such organisms show no lag phase of 
growth, and all of the data presented deal with 
transformations occurring during the logarithmic 
phase of the growth curve. 

The chilomonads used were from the Hopkins 
stock which has been maintained in sterile pure 
cultures since 1933. There have been two inter- 
ruptions in the work ; consequently the data pre- 
sented were obtained using three diiTerent samples 
of chilomonads. No differences were noted in the 
growth of the various samples. 

Numbers of cells were ascertained by fixing the 
cells in Lugol's solution and counting them in a 
hemacytometer. \\'et and dry weights were ob- 
tained by packing the cells in centrifuge tubes 
terminating in capillaries, withdrawing the super- 
natant solution, and weighing the cells before and 
after drying at 110°. Total nitrogen in the or- 
ganisms was measured either directly by aerating 



August 23, 1941 ] 



THE COLLECTING NET 



171 



Table II. 

IJ'i-f ami Dry Weights of Chilomoiwds Ihiriiu/ 

the Logarithmic Phase of Growth 

(48 Hour Cultures) 



Wet Weights 
mg./lO" cells 


Dry 
mg 


Weights 
/lO" cells 


2.64 




0.738 


2.42 




0.559 


2.42 




0.563 


2.60 




0.692 


2.51 




0.530 


2.52 




0.595 


2.48 




0.575 


2.49 




0.563 


2.87 




0.767 


2.11 




0.545 


2.51 




0.567 


2.48 


average = 


0.567 


average = 2.51 


= 0.605 


Dry weight = 


= 24% of wet 


weight. 



the ammonia from an alkalized .suspension of cells 
and suljjecting the residue to Kjeldahl digestion, 
or dififerentially by measuring the ammonia pres- 
ent Ijefore and after Kjeldahl digestion, 1)oth 
methods yielding essentially the same results. The 
micro-Kjeldahl procedure of Folin was used rou- 
tinely, and the amounts of ammonia were meas- 
ured by determining the absorption of Nesslerized 
solutions at 400 millimu with a Coleman mono- 
chromater spectrophotometer. Tests for nitrite 
were made with sulfanilic acid - a naphthylamine 
and tests for nitrate with diphenylamine and di- 
phenylbenzidine. Amide nitrogen was estimated 
by the increase in ammonia nitrogen on subject- 
ing the organisms to three-hour hydrolysis in 
0.5 N H2SO4. 

The results of the investigation are as follows : 
Table II shows the average weight of the chilo- 
monads during the rapid growth of the logarith- 
mic phase of the growth curve. Thus an average 
cell would weigh 2.5 X 10"^ mg. and would be 
24 percent solid. Table III shows the total nitro- 
gen content of the organisms, and assuming all of 
this to be protein nitrogen, an approximation 
which seems to be justified Ijy the fact that it is 
all removed by tungstic acid precipitation, the pro- 
tein accounts for about 16 percent of the wet 
weight or 67 percent of the dry weight. Inci- 
dentally the carbohydrate accounts for about 30 



percent of the solids, so these two conijjonents ac- 
count for almost all of the .solids. 

The question to be answered, however, is whe- 
ther the nitrogen in the cells accounts for all of 
the nitrogen utilized. Table IV gives the answer 
to this question. Choosing one example from 
these experiments (Experiment 2), which are all 
essentially the same, we find that the original 
total or ammonia nitrogen in the solution was 
149y/cc., that added in the organisms being in- 
significant. After 48 hours' growth 124y/cc. of 
ammonia nitrogen remained in the solution ; i.e. 
257/cc. had disappeared. Subjection of the total 
solution, i.e. solution plus organisms, to Kjeldahl 
digestion gave a final total nitrogen of 149y/cc. 
or 100 percent recovery. The population of the 
culture at this time was 4 X 10" organisms/cc. or 
1 mg. of wet cells/cc. As seen from the previous 
table, these would be expected to be 2.5 percent 
N, i.e. to contain 2Sy of nitrogen, which just ac- 
counts for the 25y of nitrogen which disappeared 
from the solution. 

We can also answer the question concerning the 
oxidation of ammonia, both indirectly by the fact 
that all of the utilized ammonia appeared as or- 
ganic nitrogen and by direct analysis for nitrite 
and nitrate. We see that no nitrate (less than 
0.025y/cc.) could be found before or after growth 
of the culture and the small increase in nitrite 
represents less than 0.1 percent of the nitrogen 
used. You will note that this experiment is the 
only one in which this increase occurred. There- 
fore in the most unfavorable experiment the in- 
crease in oxidation products of ammonia is in- 
significantly small. The conclusion therefore is 
that in the solution used Chilonwuas does not oxi- 

Table III. 

Nitrogen Content oj ChUonwnads During the 
Logaritliniic Phase oj Grozvtii 



mg. N/IO" cells 



mg. protein/lO" 
cells (N X 6.25) 



0.056 
0.071 
0.060 
0.066 
0.063 
0.066 
0.063 
0.063 

0.064 



0.40 



Nitrogen (% of wet weight) 
Protein C^A of wet weight) 



2.5% 
16% 



172 



THE COLLECTING NET 



[ Vol. XVI, No. 146 



Table IV. 
Changes in Nitrogen Content of Cultures of Chilouionas 



Expt. 


Original 

NH,-N 


Final 

NH,-N 


Final 
Total 

N 


Organic 

N 


% of 
Total N 
Fixed 


Original 
NO:-N 


Final 
NO=-N 


Original Final 
NO3-N NO3-N 


% 
recov- 
ery 




7/cc. 


7/cc. 


7/cc. 


7/cc. 




7/cc. 


7/cc. 


7/cc. 7/cc. 




1 


146 


128 


146 


18 


12.3 


0.01 


0.01 


no analysis 


100 






124 


149 


25 


16.8 


0.01 


0.03 


— — 


100 


2 


149 


124 


149 


25 


16.8 


0.01 


0.03 


— — 


100 






124 


149 


25 


16.8 


0.01 


0.03 


— — 


100 






113 


146 


?,i 


22.6 


0.01 


0.01 





100 


3 


146 


112 


146 


34 


23.3 


0.01 


0.01 


— — 


100 






112 


148 


36 


24.5 


0.01 


0.01 


— — 


101 


4 


150 


119 


149 


30 


20.0 




no analyses 




99 


5 


147 


115 


146 


31 


21.1 








98 






114 


147 


2,?, 


22.4 




no analyses 




100 






109 


149 


40 


26.9 


0.01 


0.01 





99 






110 


150 


40 


26.7 


0.01 


0.01 


— — 


99 


6 


151 


110 


150 


40 


26.7 


0.01 


0.01 


— — 


99 






110 


151 


41 


27.2 


0.01 


0.01 


— — 


100 






110 


150 


40 


26.7 


0.01 


0.01 


— — 


99 



dize significant amounts of ammonia, btit all that 
it utilizes is incorporated in organic compounds. 
Finally, to avoid leaving the impression that the 
problem is solved, I shall present the results of a 
part of the work on fractionation of the organic 
nitrogen. It would l)e of great interest to know 
just what nitrogen-containing compounds are 
made from the ammonia. Table V shows the re- 
sults of the first attempts to characterize the pro- 
teins. Analyses have been made for amide nitro- 

Table V. 
Amide Nitrogen Content of Chilomonads 



Experiment 


Organic N 
mg. 


Amide 
mg. 


N 




% 


1 


33 


11 






33 


2 


31 


8 






26 


3 


18 


5 






28 


4 


79 


20 






25 




72 


18 






25 


5 


40 


12 






30 


6 


51 


12 






24 


7 


87 


23 

average = 


32 




= 28.2 



gen. You will see that this amounts to 25-30 per- 
cent of the organic nitrogen. All of it is in the 
tungstic acid precipitable fraction. This indicates 
a high proportion of dicarboxylic amino acids in 
the proteins and I should like to point out that 
of the common proteins those from grains contain 
amide nitrogen of this order of magnitude. It 
seems to me very interesting to find this situation 
in a starch forming flagellate. 

Summary 

1. As has l)een previously reported by numer- 
otis workers ammonia is a satisfactory source of 
nitrogen for Chilomonas. It has finally been pos- 
sible to obtain sufficient growth of the organisms 
to show by direct analysis that the nitrogen found 
in the organisms came from the ainmonia in the 
culture solution. 

2. It has been shown that in the solution used 
no significant amotmts of ammonia are oxidized 
to furnish energy for growth. 

3. Finally it has been shov^-n that of the nitro- 
gen in the proteins a relatively high proportion is 
in the form of labile amide groups. Further iden- 
tification of the various nitrogen containing com- 
pounds is certainly desirable. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory or 
August 12.) 



i 



August 23, 1941 ] 



THE COLLECTING NET 



173 



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174 



THE COLLECTING NET 



[ Vol. XVI, No. 146 



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176 



THE COLLECTING NET 



[ Vol. XVI, No. 146 




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FURTHER IMPLICATIONS OF FLEXIBLE 
PROTEIN FRAMEWORKS 

Dr. Dorothy Wrinch 

Professor at Amherst, Smith and 

Mount Holyoke Colleges 

In this short report, it is my purpose to em- 
phasize two sets of imphcations following from 
the flexible protein framework picture of cytoplas- 
mic and memlirane structures. I refer to ( 1 ) the 
synthesis of proteins and (2) the function of me- 
tallic ions in physiological processes. 

( 1 ) I have suggested that long threads or rods 
of . (globular) native protein units in linear ar- 
rays, linked by multiple units to form frameworks, 
account very satisfactorily for high water/pro- 
tein ratios (Cold Spring Harbor Symposium. 
1941, forthcoming) ; may he capable also of ac- 
counting for the anomalous properties of cyto- 
plasm ; and that fabrics of such units may he the 
Ijasic structural units in biologically active mem- 
branes (Collecting Net, 16, 121, 1941). In 
such cases, the disengaged faces of the native pro- 
tein units (i.e. those not in use as interlinks of 
the frame work) are available as templates for 
synthesis and other reactions. Evidently cases in 
which high metabolic activity is associated with 
low cytoplasmic viscosity (Preston, Nature, 147, 
710, 1941) here find explanation and interpreta- 
tion, for the longer the threads, the lower the vis- 
cosity and the larger the template area. The pro- 
cess of protein synthesis may be visualized as 
taking place by means {Continued on page 190) 



NATURE, FUNCTION, AND DISTRIBU- 
TION OF THE PHOSPHAGENS IN THE 
ANIMAL KINGDOM 

Dr. O. AIeveriiof 

Research Professor of Biochemistry, 

University of Pennsylvania 

In this place, where such a variety and abun- 
dance of living material is at our disposal, I think 
it worthwhile to speak about a subject, which at 
the same time has aspects of general as well as of 
comparative phj^siology. It is true that the in- 
vestigations of the nature and distribution of the 
two phosphagens of the vertebrate and inverte- 
brate muscle had their main development in the 
years 1927-1937 and that after that date nothing 
of imjjortance was added to our knowledge. 
Nevertheless I venture to recapitulate before you 
this clear cut work on account of its bearing on 
different physiological topics. 

About the same time, in 1927, the Eggletons 
in London found a lalaile organic phosphate in 
muscle, which they called phosphagen, and Fiske 
and Subbarow in Boston discovered the same 
substance, isolated it and proved that it consisted 
of creatine and phosphate, and named it phospho- 
creatine. Then rapid progress followed ; it was 
shown that the compound was a monomolecular 
creatine phosphoric acid with a P-N linkage ; that 
the hydrolysis to creatine and phosphate was in- 
timately connected with the activity of muscle, 
and that in the recovery period it was resynthe- 
sized again. What seemed at first puzzling was 
that a great part of the creatine phosphate broken 



TABLE OF 

Nature, Function and Distribution of the 
Phosphagens in the Animal Kingdom, Dr. 
O. Meyerhof 177 

Further Implications of Flexible Protein 
Frameworks, Dr. D. Wrinch 177 

Electrical Potential and Activity of Choline 
Esterase in Nerves, Dr. D. Nachmansohn.... 182 

Chemical Composition of Mitochondria and 
Secretory Granules, Dr. Albert Claude 183 



CONTENTS 

Invertebrate Class Notes 184 

Papers and Demonstrations Presented at the 

General Scientific Meeting 185 

Book Reviews 186 

Academic Rank of M.B.L. Investigators 186 

Seminars at Mountain Lake 186 

Items of Interest 187 

Memorials Read at Corporation Meeting of 

the Marine Biological Laboratory 188 



178 



THE COLLECTING NET 



[ Vol. XVI, No. 147 



HN 



OH 

/ 

P = O 

\ 

OH 



C = NH 

N • CH3 
I 
CHo 

I 
COOH 



Creatine phosphoi-ic 
acid 



OH 

/ 

HN — P = O 

\ 

OH 

C = NH 

NH 
I 

(CH2)3 

CHNHo 

I 
COOH 

Arginine phosphoric 
acid 



down was already resynthesized anaerobically af- 
ter contraction. Since it was shown by heat meas- 
urements that the splitting was accompanied by a 
positive heat of +11000 cals pro mol, apparently 
with a similar loss of free energy, the reversal of 
this splitting had to be an involuntary endother- 
mic reaction. This puzzle could be solved very 
soon by demonstrating, that the synthesis was 
coupled with simultaneous lactic acid formation, 
which in itself is a strongly exothermic reaction. 
Under most favorable conditions two mol of crea- 
tine phosphate are synthesized for one mol lactic 
acid formed from glycogen. In this way the 
whole coupled reaction including the heat of neu- 
tralization of lactic acid is rather thermoneutral. 

Before going into more detail of the coupling 
I may describe the simultaneous development of 
this problem in the direction of comparative phys- 
iology. The Eggletons found phosphocreatine in 
all classes of vertebrate muscle, but found no 
phosphagen at all in invertebrate. The authors 
used the method to follow the time course of 
color development of molybdene blue after molyb- 
dene sulfuric acid and a reducer had been added 
to the solution in question. While with inorganic 
phosphate the intensity of the color rises steeply 
in the first minutes, the color development in 
presence of creatine phosphate is much delayed 
for 20 minutes. In this way they showed that the 
bulk of the phosphate in fresh muscles of verte- 
brates consists of the labile organic phosphate, 
called phosphagen ; by the same method they 
missed this phosphagen in invertebrate vidthout 
exception. 

In the year 1927 Dr. Lohmann and I took up 
the same problem, using muscles of crayfish. 
Since previously we had established the highly 
conspicuous role which the breakdown and syn- 
thesis of phosphocreatine plays in muscle contrac- 



tion, we were convinced that some substitute must 
exist in invertebrate muscle and by a good chance 
we came immediately on the track of it. One day 
I had left a series of acid filtrates of crayfish 
muscles on my laboratory table and preferred to 
go to lunch instead of working them up. Re- 
turning two hours later on I found to my surprise 
that by this unintentional acid incubation was lib- 
erated a similar amount of phosphate, as in frog 
muscle filtrates by the procedure of Eggleton. So 
another labile substance was present, a little more 
acid stable then phosphocreatine. The isolated 
substance proved to be arginine phosphoric acid. 

The peculiar constitution of the phosphagens, 
containing a P-N linkage, v\'as established by com- 
parison with a synthetic product, aminophosphoric 
acid. The latter behaved quite analogously with 
regard to stability, titration curve and heat of hy- 
drolysis, which amounted here to -|- 15000 cals 
pro mol. ^\'hiIe the constitution of the phospha- 
gens was established since 1928, the chemical syn- 
thesis of creatine phosphoric acid was accom- 
plished by Zeile only in 1937 and thereby the 
configuration was definitely confirmed. 

The first experiments proved the presence of 
arginine phosphoric acid in crustacean muscle 
only; but in 1928, working at the Zoological Sta- 
tion in Naples, I found it present also in repre- 
sentati\'es of annelids, molluscs and echinoderms. 
On the other hand, the acrania amphioxus, which 
is considered the immediate predecessor of the 
vertebrata, contained only creatine phosphate, no 
arginine phosphate. Therefore I spoke of a gene- 
tic "chemical mutation" of the phosphagens oc- 
curring on the level of the chordates. Indeed, ar- 
ginine can be regarded as the more primitive sub- 
stance and creatine as a special derivative of it, 
since arginine is already a component of protein. 
As we know now from Schoenheimer's work with 
isotopic nitrogen, creatine is formed in the animal 
body out of arginine, glycine and methionine. In 
contrast to arginine, creatine is an exceedingly 
stable substance and is not decomposed at all in 
the body but onl)' dehydrated to creatinine. Fin- 
ally, creatine phosphate seems better adapted to 
its biological purpose, since the heat of hydrolysis 
is 20% greater than that of arginine phosphate, so 
that the energy, useful for muscle work, is cor- 
respondingly greater. This speaks in favor of a 
biochemical improvement by substitutiiig creatine 
for arginine. 

Some years later, in 1932, Joseph Needham and 
a large group of coworkers in Cambridge, Eng- 
land, took up this problem of chenfical mutation, 
enlarging the investigations to many more classes 
of animals. They missed all phosphagen in pro- 
tozoa and coelenterates, but they found arginine 
phosphate in all phyla of invertebrates where 
muscle tissue existed. On the other hand these 



August 30, 1941 ] 



THE COLLECTING NET 



179 



animals did not contain creatine phosphate, which 
was already known for the majority of them. But 
there were two interesting^ exceptions : the hemi- 
chorda Balaiioglossus, an earlier class of chor- 
dates than the more recent acrania aniphioxus, 
contained arginine and creatine phosphate, and 
the same was true for the jaw muscles of sea ur- 
chins, the muscles in the so-called lantern of 
Aristotle. These muscles contained hoth phospha- 
gens, about twice as much arginine phosphate as 
creatine phosphate. At first sight this seemed to 
be in contradiction to the concept of chemical mu- 
tation, but Needham takes it as confirmation of 
the evolutionary theory of Bateson, who looks on 
the Echinoidea as the immediate precursors of the 
primitive chordate since both have the same kind 
of plutei-larvae. Indeed, an ontogenetic fact 
serves to support Needham's interpretation : the 
larvae themselves contain only arginine phosphate, 
while creatine appears after the metamorphosis. 
One other exception, which is not explained or 
cleared up so far, was found by a pupil of Need- 
ham in 1937: the brittle stars, belonging to the 
class Ophiuridea in the phylum Echinodermata, 
seemed to contain only creatine phosphate. The 
present state of our knowledge is given in Figure 
2 in condensed form (after Needham & Baldwin) : 
While in general no discussion arose between 
the different workers in this field owing to the 
ease, with which both phosphagens can be dis- 
tinguished one from the other and from other 
compounds, uncertainty prevailed for several 
years regarding the phosphagen of the cephalo- 
pods, like the octopus. In the first investigation 
in Naples I could not find any phosphagen in 
their mantle muscle, probably on account of the 
speed with which it decomposes in dissecting. 
Baldwin, a coworker of Needham's, claimed to 
have found a phosphagen which should differ in 
some points of arginine phosphate. Such a pos- 
sibility seemed suggestive, since Japanese authors 
had discovered in octopus muscle a basic sulj- 
stance, which proved to be a condensation product 
of arginine and propionic acid, called octopine. 
This suljstance was thoroughly investigated and 
synthesized by Prof. D. Wright Wilson in 
Philadelphia. But, as was proved by Dr. Loh- 
mann in our Laboratory, Baldwin was led astray 
by technical circumstances and the isolated phos- 
phagen of octopus proved to be normal arginine 
phosphate. This was confirmed by Dr. Wilson, 
who showed, moreover, that fresh mollusc mus- 



Phylum and class 




Arginine 
phosphate 


Creatine 
phosphate 


Most inverteljrate 


phyla 




+ 


— 


Echinodermata 










Crinoidea 






+ 





Asteroidea 






+ 





Holothuroidea 






+ 





Echinoidea 






+ 


+ 


Ophiuroidea 






— 


+ 


Protochordata 










Tunicata 






+ 





Enteropneusta 






+ 


+ 


Cephalochorda 






— 


+ 


V'ertebrata, all classes 




- 


+ 




FIGURE 


2. 







cles, of octopus or of scallop, contain mostly ar- 
ginine, and that octopine is formed post mortem, 
but probably by an enzymatic condensation, the 
significance of which is unknown. 

I shall now discuss the fact that the specificity 
of the two phosphagens extends also to the en- 
zymes concerned with their turnover. Therefore 
I must come back to the coupled reactions, in 
which the phosphagens take part. Already at the 
beginning of this paper I stated that in muscle 
activity phosphocreatine breaks down before lac- 
tic acid is formed ; that the energetic role of lac- 
tic acid formation consists in the resynthesis of 
creatine phosphate, a process of anaerobic recov- 
ery. This connection became especially clear by 
the discovery of Einar Lundsgaard in 1929 of the 
"alactacid contraction" by poisoning the muscle 
with iodoacetic acid. In such a muscle a restrict- 
ed amount of work can be done anaerobically, 
while no lactic acid is formed. At the same time 
creatine phosphate breaks down exactly propor- 
tionally to the work done and no anaerobic syn- 
thesis takes place. Lundsgaard established the 
same relation between arginine phosphate and in- 
vertebrate muscle work by poisoning the claw 
muscle of the spider crab with iodoacetic acid. 

In the years following these discoveries the 
single enzymatic reactions, by which the energy 
and the phosphate are transferred from carbohy- 
drate metabolism to creatine, were cleared up. 
Without going into details, I may state, that these 



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180 



THE COLLECTING NET 



[ Vol. XVI, No. 147 



N=C-NH2 



HC C-N 



\ 



OH OH 



CH 



OH 



N— C— N~CH— CH— CH— CH— CH — O— P— OH 

I — o — 1 ' !l 



I. Adenylic acid = Adenosinmonophosphoric acid 

N=C-NH.. 

I I 
HC C-N OH OH 

^ ! ! 

CH I I OH OH 

/ I I i I 

N— C— N— CH— CH— CH— CH— CH..— O— P— O— P— OH 

' O i " " " 



O o 



II. Adenosindiphosphoric acid 



\ 



N=C-NHo 
HC C-X OH OH 

CH i I OH OH OH 

.^11 III 

N-C— N-CH-CH-CH— CH-CH,.— O-P-O-P— O-P-OH 

O O O 



■O 



III. Adenylpyrophosphoric acid = Adenosintriphosphoric acid 

FIGURE 3. 



reactions are so called "transphosphorylations," 
where the phosphate group is transferred from 
one compound to another without becoming in- 
termediarily inorganic phosphate. All these trans- 
phosphorylations proceed by means of one single 
system, the adenylic system, -w'hich I show in 
Figure 3. It consists of three steps, discovered 
by Dr. Lohmann in the Heidelberg Institute. 
This system is to be regarded as phosphorylating 
coenzyme. Together with different proteins as 
carriers it is responsible for phosphorylations, 
dephosphorylations and transphosphorylations in 
muscle metabolism. 

The breakdown of phosphocreatine proceeds for 
instance, as Lohmann showed, in the way of equa- 
tion A and B of Figure 4. On the other hand, 
creatine phosphate is synthesized by coupling with 



two different steps of sugar metabolism. One of 
these is explained by equations C and D. The last 
phosphorylated intermediary in lactic acid forma- 
tion is phosphopyruvic acid. The latter forms 
pyruvic acid by transphosphorylation with adeny- 
lic acid. Adenosintriphosphate transfers its phos- 
phate gi'oup to creatine. 

A careful study by Dr. Lohmann revealed that 
the adenylic system is exactly the same in inver- 
tebrate as in vertebrate muscle, only the enzyme 
proteins are of different stability, and in crab 
muscle the step to adenosindiphosphate is easier 
than to adenylic acid (E). On the other hand 
the enzymes concerned with the phosphagens are 
specific : an enzymatic extract of frog or rabbit 
muscle phosphorylates creatine in presence of ade-' 
nylpyrophosphate but not arginine, and arginine 



August 30, 1941 ] 



THE COLLECTING NET 



181 



phosphate is stable in it whereas creatine phos- 
phate is split. The opposite is true for extracts 
from crayfish. Here only arginine reacts. Need- 
ham and his collaborators used this method of 
Lohmann's to reinvestigate in 1937 their former 
findings of the distribution of phosphagens in the 
muscle of Echinoderms. While in general the 
enzymatic muscle extracts of invertebrates could 
only react with arginine, the jaw muscles of sea 
urchins gave enzymatic extracts, which were able 
to phosphorylate creatine as well as arginine ; they 
contain both sets of enzymes. On the other hand 
the holothurian, also an echinoderm, which pos- 
sesses only arginine phosphate, gives enzymatic 
extracts, active towards arginine but inactive to- 
wards creatine. These experiments give a strik- 
ing confirmation of the result that in the jaw 
muscles of Echinoids, both phosphagens are pres- 
ent as active functioning suljstances. 

I may add some words about the distril^ution 
of the phosphagens in ontogenesis. According to 
Needham and his school the developing embryo 
contains an amount of phosphagen, which is 
roughly proportional to the amount of muscle tis- 
sue in the same stage of development. But 
muscle is not the only tissue to contain phospha- 
gen. Gerard and his coworkers have found phos- 
phagen in the peripheral nerves, about 1/4 as 
nuich as in the same quantity of muscle : part of 
this phosphagen Isreaks down in stimulation and 
more in prolonged asphyxia. While in mamma- 
lian nerves the phosphagen was creatine phos- 
phate, in lobster nerve, as was to be expected, it 
was arginine phosphate. 

Still more interesting is the high content of 
phosphocreatine in the electric organ of fishes. 
The percentage content in the organ of Torpedo 
is not much less than in frog muscle and related 
to the dry weight of the organ may be even 
higher. The same phosphorylating enzymes as in 
muscle extract were found in the extract of the 
electric organ. The evidence that the phosphagen 
breaks down in connection with the electric dis- 
charge is so far rather incomplete. This is an in- 
teresting question, in view of the findings of Dr. 
Nachmansohn, that the formation and the disap- 
pearance of acetylcholine are intimately connected 
with the electric discharge and that with regard 
to the concentration of cholinesterase the electric 
organ corresponds to the nerve endplate and not to 
the muscle fiber. Nevertheless, also in muscle ac- 
tivity the breakdown of phosphagen apparently is 
not the first chemical process unchained by stimu- 
lation and immediately responsible for the contrac- 
tion, but one of the consecutive steps which follow 
this unknown fundamental reaction. Therefore the 
role of creatine phosphate in the electric organ may 
be a similar one. Finally, as was found by Torres 
in the Heidelberg Institute, mammalian sperma- 



A) 2 creatinephos])hate -|- adenylic acid ^ 

2 creatine -|- adenosintriphosphate 

B ) adenosintriphosphate -> adenosindiphosphate 
+ H3PO4 -^ adenosinmonophosphate 
(adenylic ac.) +2 H3PO4 

Sa. 2 creatinephosphate -^ 2 creatine -\- 
2 phosphate 

C) 2 phosphopyruvic ac. -|- adenylic ac. — > 

2 pyruvic ac. -\- adenosintriphosphate 

D) adenosintriphosphate + 2 creatine -» 

adenylic ac. + 2 creatinephosphate 

E) adenosintriphosphate + arginine ^ 

adenosindiphosphate -|- argininephosphate 

FIGURE 4. 



tozoa possess a high content of phosphocreatine 
and a high content of enzymes, to synthesize it. 
Together with other observations about anaerobic 
glycolysis and movement of spermatozoa after 
poisoning with iodoacetic acid this observation 
favors the view that the same chemical reactions 
are responsible for the movement of spermatozoa 
as for the muscular movement. 

Summarizing our present knowledge, we may 
say, in short, that creatine phosphate developed 
out of arginine phosphate on the evolutionary level 
of the late echinoids and the early chordata and 
that both phosphagens have exactly the same 
function, which in muscle is vmdoubtedly the 
transfer of chemical energy by means of trans- 
phosphorylation and which may be similar in 
other irritable tissue. 

With growing insight into the biochemical 
structures of the cell surely more examples of 
such mutations will be discovered. So far I know 
only one other, studied by Dr. Wald. He found 
that fresh water fishes possess a kind of visual 
purple different from that of other animals, a 
visual purple, called porphyropsin, not derived 
from vitamin A, like the normal purple, Rhodop- 
sin, but from a homologue, vitamin Ao. Also here 
the physiological function of the chemically mu- 
tated substance must be essentially the same as 
that of the original substance. We may hope that 
by artificially induced chemical mutations in 
strains of virus, we may learn still more about 
the meaning of these genetic chemical transforma- 
tions. 

(This article is based upon a lecture presented at 
the Marine Biological Laboratory on August 22.) 



182 



THE COLLECTING NET 



[ Vol. XVI, No. 147 



ELECTRICAL POTENTIAL AND ACTIVITY OF CHOLINE ESTERASE IN NERVES 

Dr. D. Nach.mansohn 
Laboratory oj Physiology. Vale University School oj Medicine 



Electrical changes during nerve activity occur 
within a few milhseconds or even within a frac- 
tion of a milHsecond. Chemical reactions con- 
nected with these changes must have approxi- 
mately the same rapidity. This time factor is of 
primary importance for the theory that acetylcho- 
line (ACh) might be the "mediator" of nerve im- 
pulses across ganglionic synapses and neuromus- 
cular junctions. Such a substance must appear 
and disappear with the speed of the electrical 
phenomena. 

No data are available which establish the rate 
of ACh appearance during nerve activity. But 
the possible rate of its removal at motor end 
plates and synapses has been determined. ACh 
is inactivated by the specific enzyme choline es- 
terase. Studies on the concentration and distri- 
bution of choline esterase have revealed that at 
motor end plates and ganglionic synapses as well 
as at synapses of the C.N.S. considerable amounts 
of ACh can be split in milliseconds. These 
amounts if liberated would have a stimulating ac- 
tion. The experiments therefore indicate that the 
removal of ACh can occur at a rate rapid enough 
for the assumption that ACh is involved in the 
transmitter process. 

These results, however, do not imply that .^Ch 
is the synaptic transmitter as originally con- 
ceived. Recent investigations suggest that the 
theories of Loewi and Dale must be altered to 
account for the .\Ch inetaliolism which closely 
parallels the electrical changes occurring every- 
where at or near the neuronal surface. This new 
conception is based on two lines of observations. 

( 1 ) The first argument is based on investiga- 
tions carried out on the electrical organs of fishes. 
In spite of the great power of the discharge in 
these organs there is no reason to regard the elec- 
tricity of these organs as extraordinary compared 
with that of ordinary nerves. The organ is formed 
by electric discs or plates which are arranged in 
series. It is only this arrangement in series by 
which these organs are distinguished from other 
excitable structures and by which the high E.M.F. 
is attained. 

In strong electric organs (Torpedo and Elec- 
trophorus elcctricus (Linnaeus)) exists a high 
concentration of choline esterase. These organs 
can split in 60 minutes an amount of ACh equi- 
valent to 1-3 times their own weight. The essen- 
tial point is the fact that in these organs consider- 
able amounts of ACh can be split during the re- 
fractory period which is of the order of millisec- 
onds. This makes possible the assumption that 
ACh is closely connected with the discharge. The 



prerequisite for such a conception is the possibil- 
ity of a quick removal of the active substance. 
The high concentration of the enzyme appears 
particularly significant in view of the high water 
(927c') and low protein content (2-3%) of the 
organs. 

In the weak electric organ of rays the enzyme 
concentration is low. If in the three species num- 
ber of plates per cm. and E.M.F. per cm. are com- 
pared with the concentration of choline esterase a 
parallelism, within certain limits, is obtained. This 
parallelism has also been demonstrated, with 
Coates and Cox, on the electric organ of Electro- 
phorits electricus. If the number of plates per 
cm. and the E.M.F. per cm. are determined from 
the head to the caudal end, an S-shaped curve is 
obtained. The curve obtained for the concentra- 
tion of choline esterase is essentially the same. 
Electric organs are highly specialized in their 
function. The discharge is here the final event. 
There is no question of a transmitter function as 
there is no second unit to be stimulated. The fact 
that a specific enzyme is so highly concentrated 
in this organ — so poor in protein — is in itself sup- 
port for the assumption that the substrate is con- 
nected with its function. The parallelism found 
between intensity of discharge and activity of the 
enzyme emphasizes this relationship. 

It could moreover be demonstrated with the 
electric organ of Torpedo inartnorata. with Fes- 
sard and Feldberg. that ACh is released during 
the discharge and that injection of ACh into the 
organ produces a discharge. The discharge is 
greatly enhanced if the choline esterase is inac- 
tivated by eserine whereas eserine itself has no 
effect. 

The second line of observations leading to a 
modification of the original theories is the local- 
ization of the enzyme inside the nerve cell. Only 
a quantitative difference exists between the con- 
centration of the enzyme in nerve fibres and that 
at synapses. Experiments on the superior cervi- 
cal ganglion of cats suggested that the difference 
is related to a concentration of the enzyme at or 
near the surface of the nerve cell and therefore 
high at synaptic regions where the endarboriza- 
tion increases the surface. Direct evidence for 
this assumption was offered last year here in 
Woods Hole with Dr. Boell with experiments on 
the giant fiber of squids. It was found that prac- 
tically all the enzyme is localized in the sheath and 
that only negligible amounts occur in the axo- 
plasm. This localization indicates that ACh me- 
tabolism occurs not only at synapses but every- 
where at or near the surface. The difference is 



I 



August 30, 1941 ] 



THE COLLECTING NET 



183 



only a quantitative one. Bioelectric potentials are 
surface phenomena. The locahzation of the en- 
zyme at or near the surface is therefore particu- 
larly pertinent in view of the parallelism between 
voltage and enzyme activity. 

The observations suggest that ACh is intrin- 
sically connected with the electrical changes oc- 
curring during nerve activity at the neuronal sur- 
face. Thus the controversy between "electrical" 
and "chemical" theory of transmission of nerve 
impulses becomes meaningless. Both are signs of 
the same event. According to Eccles and Sher- 
rington and Lorente de No the excitable proper- 
ties of central neurons are similar to those of the 
peripheral axons. Gasser and Erlanger scrutiniz- 
ing the whole problem at the symposium on the 
synapse arrived at the conclusion that conduction 
of nerve impulses along fibers and across synapses 
is essentially the same process, and that the dif- 
ference is only a quantitative one. This view is 
not compatible with the theory of a specific synap- 
tic transmitter. The new conception agrees well 
with these conclusions based on the electrical 
phenomena. 

How can a relationship be pictured between 
voltage and action of ACh? According to the 



equation V=E— IR (where V is = voltage, E = 
E.M.F., I = current and R = resistance) two as- 
sumptions can l>c made alaout the way in which 
ACh may act : 

1) ACh can produce the E.M.F. directly by 
action on the surface. This possiljility appears to 
be the less probable. There is a great difference 
in concentration of strong ions between the inside 
and the outside of the nerve fiber. It seems more 
probable that these difl^erences are responsible for 
the potential dift'erences. 

2) ACh can decrease the resistance and this 
again by action on the surface, for instance by in- 
creasing the permeability. This way can be easier 
conceived. The resistance decreases during nerve 
activity, as shown by Cole and Curtiss. The ac- 
tion of ACh may be connected with this transient 
change of resistance. A substance which is re- 
leased and can be removed within a millisecond 
could well account for such a quickly reversible 
alteration of the membrane. It would be com- 
patible with the ideas on propagation of nerve im- 
pulses as developed by Keith Lucas and Adrian. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on 
August 19.) 



CHEMICAL COMPOSITION OF MITOCHONDRIA AND SECRETORY GRANULES 

Dr. Albert Claude 
The Rockefeller Institute jor Medical Research. New York, N. Y. 



The chemical nature of mitochondria, and the 
function which they assume in the economy of the 
cell, are prolalems which have failed to be solved 
by cytological techniques. The experience of the 
past fifty years leaves little hope that definite in- 
formation can be gained unless we find ways of 
isolating these cytoplasmic elements and submit 
them to direct chemical analysis and biological 
tests. Recent results indicate that mitochondria 
can be separated from the other components of the 
cell by means of a simple method of differential 
centrifugation at high speed. Under proper con- 
ditions, other particulate components of cyto- 
plasm, especially the relatively large elements 
known under the names of plasts, secretory or 
zymogen granules, can be isolated by the same 
technique. 

In previous studies, small particles, ranging 
in size between 60 and 200 nyji. diameter were iso- 
lated from normal and tumor cells. Chemically, 
these tissue particles are complexes made up es- 
sentially of phospholipids and ribonucleoprotein. 
Small particles of this type appear to l)e general 
constituents of cells and there is evidence that the 
elements purified in the high speed centrifuge 
represent mitochondria, or fragments of mitochon- 
dria (A. Claude, Science, 90, 213, 1939; Sym- 
posia on Quantitative Biology, Vol. 9, Cold 



Spring Harlrar, 1941 ; R. R. Bensley and N. L. 
Hoerr, Auat. Rec, 60. 449, 1934). In addition 
to mitochondria and "secretory" granules, the cy- 
toplasm of normal cells appear to contain also par- 
ticulate elements of smaller size which would es- 
cape detection in the ordinary microscope. 

In the present work three different kinds of 
granules were obtained from the cytoplasm of 
liver cells. Liver tissue from guinea pigs was ex- 
tracted with four times its weight of neutral 
water. Free nuclei and cellular debris were re- 
moved by a preliminary centrifugation at low 
speed. Further fractionation of the liver extract 
was carried out in a high speed centrifuge, under 
a uniform centrifugal force of 18,000 X gravity. 
The first fraction was brought down by a run of 
exactly 5 minutes at high speed. Under the mi- 
croscope, this purified fraction was found to con- 
sist of small spheres of various sizes, ranging ap- 
proximately from 0.5 to S/x, and resembling fat 
globules. On chemical analysis, these granules 
were found to contain 12 per cent nitrogen, 0.9 
per cent phosphorus, and 53 per cent carbon. 
Twenty-two to 24 per cent of the material was 
soluble in alcohol and chloroform. In size and 
appearance, the large granules correspond to the 
"secretory" granules, or plasts, which can be 
demonstrated in the cytoplasm of the hepatic cell 



184 



THE COLLECTING NET 



[ Vol. XVI, No. 147 



by proper cytological techniques (R. Noel, Arch. 
Anat. Micros., 19, 1, 1923). 

The second fraction was sedimented by 45 to 
60 minutes centrifugation at high speed. The 
purified material was a jelly-like substance, color- 
less and entirely transparent. It was composed 
of small particles visible, under dark-field illumin- 
ation, as dense, refractile bodies, spherical in 
shape, or slightly elongated. The results of chem- 
ical analysis were 9 per cent nitrogen, 1.2 per cent 
phosphorus and 56 per cent carbon. Forty to 45 
per cent of the material was soluble in organic 
solvents. Physically and chemically, the second 
fraction corresponds to the usual ''mitochondria" 
fraction referred to above. The third fraction was 
sedimented by a run of two hours at high speed. 
The purified pellet was a perfectly transparent 
mass, cherry-red in color. In the dark-field mi- 
croscope, a suspension of the material was found 
to be composed of small particles which, from 
their sedimentation rate, appear to range in size 
between 40 and 60 ni/j, diameter. A ribose nu- 
cleic acid was isolated from both the first and the 
second fractions. This nucleic acid was identified 
by characteristic absorption spectrum in the ultra- 
violet, and typical color tests. 

The above observations indicate that the "secre- 
tory" granules of the guinea y)ig liver, like the 
small tissue granules, are complex formations 
made up of lipids and proteins. Both the secre- 
tory granules and the mitochondrial material con- 
tain a ribose nucleic acid in aliout the same pro- 
portion. Both elements are equally sensitive to 
acid. Slight acidification of the medium causes 
rapid agglutination of the granules or the parti- 
cles, and in both cases, a point of minimum solu- 
bility is found at pH 3.5. The differences brought 
out by elementary analysis are of a quantitative 
nature, and. to a great extent, result from the fact 
that the large granules contain 20 per cent lipids, 
against 40 to 45 per cent for the small particles. 



The similarity in chemical constitution and be- 
havior between the large granules and the small 
particles suggests a possible continuity between 
these cytoplasmic elements. The findings may 
give support to the view that mitochondria devel- 
op into secretory granules, as suggested for the 
liver by Noel (Arch. Anat. Micro's^. 19, 1, 1923). 
If this is the case, we should be able to isolate 
forms of transition between mitochondria and ma- 
ture secretory granules. This point will be the 
matter of further work. 

An important problem intimately connected 
with the chemical nature of the cytoplasmic gran- 
ules is the role they may play in cellular physiol- 
ogy. In this respect, it may be significant that 
iron and copper are found in relatively large 
amounts in secretory granules and mitochondria. 
The particles isolated from the dried cells of 
Brewer's yeast had a copper content of 0.116 per 
cent. In this case, the purified pellet presented 
a definite blue color, resembling that of hemocy- 
anins. Similar fractions from other sources were 
found to contain copper also, but in lesser 
amounts. The proportion of copper was 0.023 
per cent for the particles derived from a mouse 
leukemia, 0.016 per cent for the small particles 
of the guinea pig liver, and as much as 0.034 per 
cent for the secretory granules. This latter quan- 
tity is not negligible if we consider that it repre- 
sents approximately 20 per cent of the copper 
content of the hemocyanin of Limulus (A. C. 
Redfield, Biol. Rcviccvs. 9. 175, 1934). There is 
evidence tthat the copper present in the granules 
occurs in combination with a protein. The pos- 
sibility that the copper compound, associated with 
the phospholipoid-ribonucleoprotein complex, may 
act as a catalyst of respiration is being investi- 
gated. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on 
August 19.) 



INVERTEBRATE CLASS NOTES 



The study of Echinoderms and Arthropods, 
two field trips, beach parties, and a baseball game 
were the high lights of another "typical" week for 
the Inverts. 

The behavior of starfish, the pretty if complex 
"Aristotle's Lantern" of the sea urchin, regenera- 
tion in the sea cucumber, particularly held our in- 
terest. Dr. Bissonnette's lecture covered the tax- 
onomy of the Echinoderms, their unique organ 
systems, their strange development. Dr. Martin 
has been following a similar plan in presenting 
the Arthropods. In addition to the lobster and 
crab, we have been studying specimens of local 
Crustacea, autotomy in the fiddler crab and its 
color reactions. 

The field trips to Hadley Harbor and North 
Falmouth were most fruitful, both in view of the 



number and variety of specimens. The exhibit in 
the lobby of the Brick Building will testify to 
that. The North Falmouth trip was a chilly one. 
Even the shoulders could scarcely keep warm. 
The hot coffee went quickly at lunch. The after- 
noon was warmer, the collecting, appropriately 
for the last trip, was the best ever. 

Our versatile group displayed their talents at 
the "Four O'Clock Club" in Falmouth. Julie and 
"Stubby" Rankin thrilled the audience with their 
dancing. Not to be outdone. Jack Osmun, Sid 
Pond, Howie Miner and Bob Corder sang their 
favorites, after which John led the enthusiastic 
patrons in group singing. 

The Staff-Invert baseball game was great fun. 
Dr. Martin is some pitcher. The game was close 
(Continued on page 191) 



August 30, 1941 ] 



THE COLLECTING NET 



185 



PAPERS AND DEMONSTRATIONS PRESENTED AT THE GENERAL SCIENTIFIC 

MEETING, 1941 



Tuesday, August 26, Morning Session, 9:00 A. M. 

Caswell Grave: Further studies of metamorphosis 
of Ascidian larvae. 

Caswell Grave: The "Eye Spot" and light re- 
sponses of the larva of Cynthia partita. 

Llovd Birmingham: Regeneration iii the early 
zooids of Amaroucium, constellatum. 

Ivor Corniian : Characteristics of the acceleration 
of Arbacia egg cleavage in hypotonic seawater. 

Ethel Browne Harvey: Material inheritance in 
Echinoderm hybrids. 

E. S. Guzman Barron and J. M. Goldinger: Inter- 
mediary carbohydrate metabolism of sperm and eggs of 
Arhacia before and after fertilization. 

J. D. Crawford, D. Benedict, A. B. DuBois and A. 
E. Navez: On contraction of the Venus heart. 

Alfred M. Lucas and James Snedecor: Coordina- 
tion of ciliary movement in the Modiolus gill. 

LORUS J. Milne: Preparing an animated diagram of 
somatic mitosis. 

Tuesday, August 26, Afternoon Session, 2:00 P. M. 

E. Newton Harvey : Stimulation by intense flashes 
of ultra violet light. 

T. C. Evans, G. Failla, J. C. Slaughter and E. P. 
Little : Influence of the medium on the radiosensitiv- 
ity of Arhacia sperm. 

C. Ladd Prosser and G. L. Zimmerman: Compara- 
tive pharmacology of myogenic and neurogenic hearts. 

Lois E. TeWinkel: Structures concerned with yolk 
absorption in the dogfish, Squaliis acanthias. 

W. H. F. Addison: The distribution of elastic tissue 
in the arterial pathway to the carotid bodies in the dog. 

Richard G. Abell and Irvine H. Page: Behavior of 
the arterioles in hypertensive rabbits, and in normal 
rabbits following injection of angiotinin. 

Wednesday, August 27, Morning Session, 9:00 A. M. 

M. H. Jacobs and Dorothy' R. Stewakt: Catalysis 
of ionic exchanges by bicarbonates. 

Dorothy R. Stewart and M. H. Jacobs: The role 
of carbonic anhydrase in the catalysis of ionic exchanges 
by bicarbonates. 

Martin G. Netsky and M. H. Jacobs: Some effects 
of desoxycortico-sterone and related compounds on the 
mammalian red cell. 

Herbert Shapiro and Hugh Davson: Permeability 
of the Arhacia egg to potassium. 

A. K. Parpart: Lipo-protein complexes in Arhacia 
eggs. 

S. E. Hill: Relation between the action potential 
and protoplasmic streaming in Cliara and Nitella. 

Aurin M. Chase: Observations on luminescence in 
Mnemiopsis. 

Papers Read by Title 

Fred W. Alsup: Photodynamic studies on Arbacia 
eggs. 

Ivor Cornman : Disruption of mitosis in Colchicum 
by means of colchicine. 

T. C. Evans: The effect of roentgen radiation on 
tlie jelly of the Nereis zygote. 

R. Ruggles Gates: Tests of nucleoli and cytoplas- 
mic granules in marine eggs. 

Russell P. Hager: Sex-linkage of stubby (sh) in 
Hdbrohracon. 

E. R. Hayes: The Elasmobranch interrenal; a pre- 
liminary note. The interrenal body of Alopias vulpinus 
(Bonnaterre). 

DwiGHT L. Hopkins : The cytology of Amoeha ver- 
rucosa. 

George W. Hunter, III, and Edward Wasserman : 



Observations on the mealanophore control of the cunner 
Tautugolahrus aclspersus (Walbaum). 

Florence Moog: The influence of temperature on 
reconstitution in Tuhularia. 

Clinton M. Osborn : Factors influencing the pigmen- 
tation of regenerating scales on the ventral surface of 
tlie summer flounder. 

G. H. Parker: Hypersensitization of catfish melano- 
phores to adrenaline by denervation. 

Leonard P. Sayles: Implants consisting of young 
buds, formed in anterior regeneration in Clymenella, 
plus the nerve cord of the adjacent old part. 

A. A. Schaepper: Chaos nohilis Penard in perma- 
nent culture. 

Victor Schechter: Further studies in Mactra egg 
cells. 

Sidney F. Velick: The effect of eeutrifngation upon 
the oxygen consumption of Arbacia eggs. 

Allyn Waterman: Ectodermization of the larva of 
Arhacia. 

Ralph Wichterman: Studies on Zoochlorella-iree 
Paraiiiecium hursaria. 

Floyd J. Wiercinski : An experimental study of in- 
tracellular pH in the Arbacia egg. 

E. Alfred Wolf, Marton Dytche and Milton 
Schafpel : Heat produced by respiring whole blood of 
Tautoga oidtis and Mtistelus canis. 

C. L. Yntema : Effect of differences between stages 
of donor and host upon induction of auditory vesicle 
from foreign ectoderm in the salamander embryo. 

Wednesday, August 27, 2:00-4:00 P. M. 
Demonstrations 

L. P. Boss and M. H. Jacobs: Stabilized source of 
current for lamps and other purposes. 

E. R. Clark and Eleanor Linton Clark: Behavior 
of giant cells as observed in the living mammal. 

E. X. H.ARVEY AND F. J. M. SiCHEL: Apparatus for 
intense flashes of ultra violet light, and the killing of 
small organisms. 

E. N. Harvey and F. J. M. Sichel: Apparatus for 
higli speed pliotography with the microscope. 

Kurt G. Stern ; An air-driven high-speed centrifuge 
for optical observations. 

Ivor Cornman: Disruption of mitosis in Colchicum 
by colchicine. 

Sears Crowell : Nematostella, a simple anemone, 
suitable for laboratory work in general zoology. 

Sears Crowell: The nematosomes of Nematostella. 

John Keosian : Apparatus of simple construction 
for use in microchemical work. 

John Keosian : A spot test method for the quantita- 
tive determination of magnesium in tenths of a micro- 
gram. 

Eleanor H. Sliper: A mutant Drosophila melano- 
gastcr with extra sex combs. 

Lois E. TeWinkel: Structures concerned with yolk 
absorption in the dogfish. 

Dr. Eric Loewenstein: Demonstration of fluoro- 
photonieter, and discussion of fluorometric methods of 
determining biological substances. 

Dr. Laurence Irving. Dr. P. F. Scholander, Mr. C. 
Lloid Clapp, Mr. George Edwards, Mr. Niels Hau- 
gaard : Section A : Sensitive volumetric apparatus de- 
vised by Dr. P. P. Scholander, as used in the continuous 
measurement of respiration and in the measured delivery 
of small quantities of liquid. Section B: A system suit- 
able for aquatic animals, sensitive to 0.02 cc, used to 
measure the Oj consumption of fishes of 20-200 grams 
body weight at 10 minute intervals for periods lasting 
for 12 to 24 hours. 



186 



THE COLLECTING NET 



[ Vol. XVI, No. 147 



The Collecting Net 

A weekly publication devoted to the scientific work 
at marine biological laboratories. 

Edited by Ware Cattell with the assistance of 
Boris I. Gorokhoff and Judy Woodring. 

Entered as second-class matter, July 11, 1935, at 
the U. S. Post Office at Woods Hole, Massachusetts, 
under the Act of March 3, 1879, and re-entered, 
July 23, 1938. 



BOOK REVIEWS 

HANNUM, C. A. "Comparative Chordate Anatomy." 
pp. vii + 211. Stanford University Press. 1941. 
$2.00. 

This is one of the numerous texts of compara- 
tive anatomy, based on the author's own course, 
which have appeared in the last year : in the opin- 
ion of the reviewer, it is better than most. Too 
many texts of this type stop short at birds, on the 
ground that any mammal, all too frequently the 
cat, should be the subject of a separate semester's 
work. Hannum has the courage to put the mam- 
mal where it Ijelongs : in the last chapter, not in 
the next book. This makes it possible to give a 
one year survey of the animal kingdom, offering 
the student an adequate background for his subse- 
quent specialised studies — among them, should it 
be desirable, the cat. 

This book not only descrilies the dissection of 
the usual laboratory types in clear language but 
also discusses classification, phyletic origins and 
ontogenesis. /Ml anatomical terms are italicised 
and the author holds a hapjjy lialance between the 
English and Latin tongues. The aljsence of illus- 
trations and the paj^er binding are not defects for 
they keep the price within the limit which any 
student can be asked to pay : and for his money 
he will get heavier ]iaper and clearer i^rinting than 
he might expect. 

There is no doubt that this text deserves care- 
ful consideration from every teacher who is plan- 
ning next year's course in comparative anatomy. 

Peter Gray 

STILES, K. A. "Handbook of Microscopic Charac- 
ters of Tissues and Organs." pp. vi + 148. Phila- 
delphia, The Blakiston Company. 1940. $1.50. 

This work endeavors to apply to the identifica- 
tion of tissues the principles used by taxonomists 
in the construction of "keys". It is very doubt- 
ful whether anyone not intimately acquainted with 
the taxononi}' of a group has ever successfully 
identified a species from a key: certainly the stu- 
dent using this book will require the help of a 
competent and willing instructor. In his Preface 
the author states that the book "is actually an ab- 
breviated text which contains . . . the fundamen- 
tals of regular histology textbooks, but in a form 
much more easily and quickly grasped by the 
reader" : that "to the student preparing for State 



and National Board Examinations, it is invalu- 
able" : that it "serves primarily to alleviate the 
welter of confusion which arises when one is pre- 
sented with a mass of facts and not enough basic 
knowledge of the subject matter to make an in- 
telligent choice". The reviewer cannot go all the 
way with the author in his opinion of the text but 
it will doubtless be found useful by those who de- 
sire a condensed concomitant to their regular lec- 
ture and laboratory volumes. 

The few illustrations for which space has been 
found are excellently drawn and the tabular sum- 
maries which preceed each section could not be 
improved. The book is ring bound with a water- 
proof cover and interleaved with blank sheets. 

Peter Gray 



ACADEMIC RANK OF M. B. L. INVESTIGATORS 

The number of investigators of each academic 
rank registered (filled out blanks before August 
21) at the Marine Biological Laboratory: 

Professors 66 

Associate Professors 23 

Assistant Professors 42 

Instructors 35 

Research Associates 7 

Assistants 55 

Fellows 20 

Graduate Students (not listed elsewhere) 37 

Medical Students 5 

Undergraduate Students 8 

Preparatory Students 5 

Not falling in above categories 26 



SEMINARS AT MOUNTAIN LAKE 

Recent speakers at the seminars, with their 
topics, at the Mountain Lake Biological Station 
have Ijeen : 

Grace T. Wiltshire, "Nesting Habits of Alleghan- 
ian Warblers." 

Edward M. McCrady, Jr., "Physiology and Embry- 
ology of the Opossum," and "Leonardo de Vinci as 
an Anatomist." 

Robert K. Burns, "Further Notes on the Embry- 
ological Development of the Opossum." 

H. Eugene Brown, "Progress Report on Termite 
Investigations." 

T. S. Painter, "Salivary Chromosomes." 



CURRENTS IN THE HOLE 


At the following 


hours 


(Daylight Saving 


Time) the current i 


n the 


Hole turns to run 


from Buzzards Bay 


to Vineyard 


Sound: 


Date 




A.M. 


P.M. 


August 31 .... 




11:39 




September 1 .. 




12:24 


12:42 


September 2 .. 




1:25 


1:43 


September 3 .. 




2:23 


2:39 


September 4 .. 




3:14 


3:30 


September 5 .. 




4:01 


4:18 






4:42 


5:00 


September 7 .. 




5:23 


5:42 


September 8 .. 




6:05 


6:25 


September 9 .. 




6:45 


7:09 



August 30, 1941 ] 



THE COLLECTING NET 



187 



ITEMS OF INTEREST 



The invertebrate course of the Marine Biologi- 
cal Laboratory is holding its last session this 
morning. 

Dr. Colin M. MacLeod, associate of Rockefel- 
ler Institute, has been appointed director of the 
Bacteriological Laboratories at the New York 
University College of Medicine. 

Dr. Frank H. Connell has been promoted 
from assistant professor to full professor of zoool- 
ogy at Dartmouth College, and Dr. James F. 
Crow has been appointed instructor there. 

Dr. Samuel R. M. Reynolds has been ap- 
pointed research associate in the department of 
embryology of the Carnegie Institution of Wash- 
ington (Baltimore). He has been associate pro- 
fessor in physiology in the Long Island College 
of Medicine. 

Recent appointments at Harvard University in- 
clude those of Drs. Paul J. Allen and Roger W. 
Sperry as research fellows in biology. Dr. An- 
thony O. Dahl has been promoted to faculty in- 
structor in biology and tutor. 

Mr. William H. Burt has been promoted 
from instructor to assistant professor of zoology 
at the University of Michigan. He is also cura- 
tor of mammals in the Museum of Zoology. 

Misses Priscilla Anderson and B. Elizabeth 
Horner have been promoted to instruciorships in 
zoology at Smith College. 

Dr. Elso S. Barghoorn, Jr. of Harvard Uni- 
versity has been appointed instructor in the biol- 
ogy department of Amherst College. 

Mr. Sidney M. Pond, who graduated from 
Wesleyan this spring, and is also in the inverte- 
brate zoology course, has been appointed graduate 
assistant in biology at Wesleyan. 

Miss Juanita Senyard, who graduated from 
Oberlin College in June, has been appointed 
graduate assistant in histology at Mt. Holyoke 
College. 

M.B.L. TENNIS CLUB 

In the finals of the ladies' singles, D. Baitsell 
defeated Mary Chamberlain, 6-2, 6-0. Stunkard 
and Evans won over Krahl and Lancefield in the 
finals of the men's doubles, 7-5, 6-2. Gary Col- 
ton defeated Huntington Mavor in the junior 
singles, 6-2, 6-3. A cup was presented to the 
winner of the junior singles by Dr. D. E. Lance- 
field. The winners of the other tournaments will 
have their names engraved on cups owned by the 
Club. 



Recent visitors at the Laboratory include Drs. 
H. J. Muller, Gertrude Gottschall. Oscar Bodan- 
sky, Elvira de Liee, Benjamin C. Gruenberg, 
Hans Gaffron, R. Leuchtenbcrger. R. Schoen- 
heimer, and R. Beutner. 

Among the new investigators who have recently 
come to the Laboratory are Dr. Robert Bloch of 
Yale University, who is doing research work in 
the library, and Miss Pauline Sullivan, teacher of 
biology and chemistry at the Choate School, 
Brookline, Massachusetts, who is assisting Dr. B. 
H. Grave in histological work. 

Several men at the Laboratory have recently 
been called up by their respective draft boards : 
Lauren C. Gilman went into service at Camp 
Devens several weeks ago ; Robert Spier leaves on 
September 1 5 for Camp Devens ; H. Duncan Rol- 
lason, Jr., left last week for Connecticut to take 
his physical examination. 

Mr. Elmer Higgins, chief of the Division of 
Incjuiry respecting Food Fishes of the U. S. Fish 
and Wildlife Service will visit the Fisheries sta- 
tion in Woods Hole early in September. Mr. 
Higgins was director of the Bureau of Fisheries 
Station here for several years. 

Dr. Gerald W. Prescott, who has been on the 
staff of botany course at the Marine Biological 
Laboratory for several summers, taught courses 
in aquatic flowering plants and the taxonomy of 
the fresh water algae at the biological station of 
the University of Michigan this summer. 

Dr. Leonard I. Katzin. who worked at the 
Marine Biological Laboratory last year, is at 
]3resent with the U. S. Public Health Service, as- 
signed to the Southeast Health District of the 
State of Virginia. His assignment is on a mos- 
quito control program. 

M.B.L. CLUB NOTES 

The M.B.L. Club House will remain open until 
approximately September 18. Tonight's dance is 
to be the last of the season. 

Miss Margaret Mast is the new chairman of 
the house committee of the M.B.L. Club. The 
new chairman of the social committee is Mrs. 
Shirley D. Hobson. 

In the ping pong tournament Dr. John O. 
Hutchens was the victor in the men's singles, de- 
feating Richard Byrrum, and will have his name 
engraved on the trophy paddle. In the finals of 
the ladies' singles, Katya Zarudnaya defeated 
Marion Davis. 



188 



THE COLLECTING NET 



[ Vol. XVI, No. 147 



IN MEMORY OF DECEASED MEMBERS OF THE CORPORATION OF THE 
MARINE BIOLOGICAL LABORATORY 



Memorials Adopted at the Annual Meeting of the Corporation, August 12, 1941 



DAVID HILT TENNENT 

In the premature death of David Hilt Tennent, 
the Corporation of the Marine Biological Labora- 
tory has lost the crowning years in the life of a 
mem1)er distinguished for his accomplishment in 
research and even more for his quality as a man. 
Tennent was an investigator who proceeded with- 
out haste, vet unceasingly ; for him quality not 
quantity of publication was the prime considera- 
tion, yet the volume of his published work is im- 
pressive. He was honored for his work by the 
Presidency of the American Society of Zoologists 
and by similar offices, and most notably by elec- 
tion to the National Academy of Science. He 
made outstanding contributions in his studies 
upon hybridization, fertilization and egg organiza- 
tion in Echinoderms, and in his later research 
upon photosensitization in which he took especial 
satisfaction since he regarded it as the most im- 
portant work of his life. These contributions, 
which are familiar to workers in these fields, are 
not so well knjwn tj many investigators in other 
lines, because lennent was the most modest and 
retiring of men. He had none of the flare for 
self-advertisemer.t that carries some men so far 
on a m.Kiicum of v orth. His every publication 
was marked not only by the critical nature of his 
observations and experiments but also by the 
meticulous care witn which each phrase was 
weighed to make sure it meant exactly what he 
had in mind, no more and no less. What he 
wrote or said publicly was always as exact as he 
could make it. Knowing the quality of the man 
and of his mind, I think one may feel that what 
he did is likely to stand until it becomes obsolete 
with the ad\'ance of knowledge, as so often hap- 
pens although the historical importance of the 
work remains. 

In his work as a teacher of undergraduates and 
as a director of graduate students, Tennent was 
no less effective. The same thoroughness and de- 
termination to do his best characterized his teach- 
ing as it dil his research. Tennent and I were 
gradratc stu.lcnts t igcther at the Johns Hopkins 
and fellow members of Drew's Invertebrate staff 
at the Marine Laboratory. I well recall his first 
lecture to the Invertebrate class. He was so 
scared the chalk rattled against the board, but he 



did better than he thought and after that first 
summer the rest of us felt we must keep up with 
him. At the Hopkins he was not satisfied with 
his first year's seminar lectures. To be safe the 
next year, as he confided to me later, he went to 
the laboratory the evening before each lecture, 
turned on the lights in the empty seminar room 
and put himself through a dress rehearsal of his 
lecture to be given the next morning. It was this 
kind of determination and performance that char- 
acterized all his work. It had to be done as well 
as he could do it. I was told that E. A. Andrews 
went so far as to say at the time that Tennent 
was the best assistant he had ever had. Only 
those of us who were assistants to Andrews can 
fully appreciate what that meant as to quality of 
performance. Thus, Tennent had the instinct of 
workmanship at the beginning of his career. 

Tennent always commanded the loyalty and 
admiration of graduate students to a marked de- 
gree. His great disappointment was that he did 
not train more students who were able to find 
places commensurate with their ability. Those in 
his confidence knew that he often longed for a 
position \vhere he might have had more "disci- 
ples". He trained manj' women of ability, but 
for the most part, in this man's world, they could 
not find positions worthy of their competence. His 
summers at the Tortugas Laboratory gave him 
opportunities to extend the kind of contacts he 
might have had in larger measure throughout the 
year in some institutions. I have often heard of 
what a stimulus he was to the younger investiga- 
tors at Tortugas summer after summer. 

On the personal side, Tennent was always quiet 
and reserved, though in his later years as well as 
in his youth a delightful companion to those who 
knew him well. He may have seemed austere to 
those who knew him casually. Yet he had a keen 
.sense of humor for all his quietness. I never 
knew a man whose sense of obligation to do what 
he thought just and right seemed to me stronger 
nor a man whom I would trust further. Thinking 
of him personally, one felt that here was a man to 
whom the abused and meaningless phrase "a gen- 
tleman and a scholar" might be applied with 
meaning. 

To Mrs. Tennent and to his son, who as a 



August 30, 1941 ] 



THE COLLECTING NET 



189 



scientist follows in his father's footsteps, we ex- 
tend our deepest sympathy. 

W. C. Curtis 

EDWARD BROWNING MEIGS 

Edward Browning Meigs was born in Philadel- 
phia September 10, 1879, the son of Arthur Vin- 
cent Meigs and Mary Roberts Browning. He be- 
longed to an old and distinguished American fam- 
ily of South English ancestry, and was a direct 
descendant of Vincent Meigs who came to Amer- 
ica and settled in New Haven, Connecticut, in 
1644. His father, grandfather, and great grand- 
father were physicians ; his great great grand- 
father, Josiah Meigs, was professor of mathema- 
tics and natural philosophy in Yale University in 
the 1790's. Scientific interests were strong in his 
ancestry. His father, a pediatrician, was inter- 
ested in the chemical composition of milk and pub- 
lished papers on this subject ; he also introduced 
a method of modifying cows' milk to make it suit- 
able for infants. In Edward Meigs' memoir of 
his father he "finds it difficult to say whether 
more of my father's energy was devoted to the 
practice of medicine or to research". 

Edward Meigs was the fourth physician in his 
family in the direct line, but his own interests 
were primarily scientific and he did not engage in 
practice. He graduated from Princeton Univer- 
sity in 1900 and took his M.D. at the University 
of Pennsylvania in 1904. He was Assistant in 
Physiology in the same University during 1904- 
1906 and then spent a year abroad in study and 
research, working chiefly in Jena with the com- 
parative physiologist Wilhelm Biedermann. He 
also spent some time in Cambridge University, 
chiefly in association with Walter Fletcher and 
Gowland Hopkins, whose work on the physiology 
and biochemistry of muscular contraction was of 
special interest to him. He was instructor of 
physiology in the Harvard Medical School during 
1907-10. In 1910 he joined the Wistar Institute 
in Philadelphia in order to devote himself entirely 
to research. From 1915 until his death he was 
physiologist in the Bureau of Animal Industry of 
the U. S. Department of Agriculture. 

He first attended the Marine Biological Labora- 
tory in 1904, and was elected a member of the 
Corporation in 1905. During the four summers 
of 1912-1915 he served as instructor in the physi- 
ology course. He and his family have been sum- 
mer residents of Woods Hole for a period of 
about thirty years, and his interest in the Labora- 
tory has been constant. 

In 1910 he married Margaret Wister of Phila- 
delphia who with two sons and two daughters sur- 
vives him. He died November 5, 1940, after a 
prolonged illness. 

Edward Meigs' early investigations were in the 



field of general physiology, especially the physiol- 
ogy of muscular contraction. Later, after he went 
to Washington, his work had reference chiefly to 
the physiology of milk production and related 
topics ; problems connected with administration 
and the organization of research in this field also 
engaged much of his attention. 

His early studies on the comparative histology 
and biochemistry of smooth and striated muscle, 
both vertebrate and invertebrate, were varied and 
extensive. His photographs of striated muscle 
fibres under high magnification are among the 
best that we have. His belief that changes of ten- 
sion in muscle were a result of reversible changes 
of hydration in the fibrils led him to experiment 
on the influence of variations of osmotic pressure, 
chemical conditions and temperature on the water 
content and correlated state of contraction of dif- 
ferent types of muscle. At one time he was 
greatly interested in physical models of muscular 
contraction, especially McDougall's model, in 
which increase of volume of inflation resulted in 
shortening. His interest in the relation of inor- 
ganic salts to contraction (as shown e.g., in the 
potassium contraction of striated muscle) led him 
to make comparative analytical studies of the salt 
content of smooth and striated muscle, vertebrate 
and invertebrate. This work had an indirect but 
important bearing on his later work in the De- 
partment of Agriculture on mineral metabolism in 
its relation to milk production. He recognized 
that the selective action of the muscle cell in ac- 
cumulating its highly special salt content had the 
same physiological basis as the selective separa- 
tion of salts by the mammary gland in milk secre- 
tion. Problems of permeability also interested 
him in relation to both the properties of muscle 
and the processes of secretion ; and in work on 
artificial membranes he showed that impregnation 
of collodion films with insoluble calcium and mag- 
nesium salts formed membranes approaching liv- 
ing plasma membranes in their semipermeability, 
a property which in the living cell also is de- 
pendent on calcium. He found, however, that the 
behavior of smooth muscle in anisotonic Ringer's 
solution and sugar solution differed from that of 
striated muscle and indicated the presence of a 
much less diffusion-proof surface layer. This dif- 
ference in physical properties he correlated with 
other evidence of a fundamental difference in the 
mechanism of contraction in the two types of 
muscle. 

During Edward Meigs' work of twenty-five 
years in the Department of Agriculture he and 
his associates made varied and important contri- 
butions to the physiology of milk production. 
Mineral metabolism and vitamin supply in rela- 
tion to milk production received special attention, 
and the results of this work were published in a 



190 



THE COLLECTING NET 



[ Vol. XVI, No. 147 



long succession of special papers and reviews. He 
was also responsible for the general planning and 
direction of the work at the experimental farm at 
Beltsville, Maryland. Many problems of a highly 
practical kind also came up for consideration ; for 
example the incidence of mastitis in the experi- 
mental herd led to an investigation of the pathol- 
ogy of this condition and effective methods for its 
control were developed, including modification of 
certain types of milking machine which v^-ere found 
to be largely responsible. The work of these years 
is too varied to summarize briefly ; some of its 
practical results are seen in the progressive ini- 
])rovement during recent years in the general 
methods employed in the dairy industry. 

Much of Edward Meigs' success in this work 
came from the thoroughness, objectivity and free- 
dom from bias that were characteristic of his 
scientific activity and outlook. He was highly 
tenacious in his convictions once thev were formed. 



but his conclusions were always based on a clear- 
sighted and critical consideration of evidence, in 
the collection of v^hich he spared no pains. Al- 
though weakened in his later years by illness, he 
maintained his scientific interest and activity to 
the last. His personal interests other than scien- 
tific were remarkably wide ; he was fond of na- 
ture and outdoor life, a great sailor, a man of 
imagination and culture, widely read, modest, 
loyal, highminded and devoted to his friends. His 
characteristic generosity was well shown in his 
gift to the Marine Biological Laboratory in 1936 
of the bathing beach property, including the bath 
house, on the Buzzards Bay front adjoining his 
family summer cottage. The use of this beach by 
members of the Laboratory, as well as of the ten- 
nis courts which occupy part of his property is 
thus permanently assured. 

R. S. LiLLIE 



FURTHER IMPLICATIONS OF FLEXIBLE PROTEIN FRAMEWORKS 

(Continued from page 177) 



of intermediates, formed by condensations of 
suitable materials adsorbed on individual pro- 
tein faces. This idea fits with the more de- 
tailed picture now emerging from enzyme 
studies, according to which there is a division 
of labor among the enzymes concerned in 
such syntheses, each enzyme catalyzing the con- 
densation of certain building blocks only (Fruton, 
Cold Spring Harbjr Syniposium, 1941, forthcom- 
ing). Such a situation explains the existence of 
symmetric arrangements of R-groui:>s such as are 
known to he present in insulin, for examj^le It 
also fits with the existence of precursors such as 
Svedberg's prolactalbumin and several other pro- 
tein intermediates, ivhich curiously enough all 
have molecular weights in the neighborhood of a 
thousand, corresponding possibly to individual 
faces or other precisely defined fragments of 72- 
residue or 288-residue skeleton cage structures. 
Further, the protein frameworks offer, not only 
characteristic and specific individual protein faces 
as templates for the formation of these interme- 
diates but, in the branch point protein units, pro- 
vide nests where such platelet intermediates ma}- 
find themselves in favorable juxtaposition for the 
final synthesis into complete cage molecules. In 
suggesting that such branch points in protein 
frameworks are the seat of protein synthesis, it 
may also be pointed out that the adsorption in 
such nests of a foreign molecule, e.g.. any antigen, 
will i'l g-Teral provi'-'e templates of new specifi- 
cities, leading to the formation of proteins of spe- 
cificities complementary to those of the foreign 
molecule, e.g., antibodies. These pointers regard- 
ing protein intermediates and the mechanism by 
which a system imposes certain specificities on 
them could be utilized in the design of experi- 
ments. 



(2) As our central and fundamental postulate 
it has been assumed that the proteins in such 
structure systems are in their native form. It 
follows that the interlinking of such units into 
frameworks also shows specificity. In connection 
with certain particular cases of one of the three 
types of such interlinkings, namely the riveting 
together of ionized groups by some divalent ions, 
certain suggestions regarding the function of such 
ions emerge. 

The fact that young tomato plants grown in 
pyrex glass containers with nutrient solutions 
deficient in zinc give a measurable and reproduci- 
ble response to the addition of one gamma of zinc 
to a plant ( giving a zinc concentration in the cul- 
ture solution of 5 parts per billion) may be taken 
as typical of many facts regarding the importance 
of minute concentrations of certain elements 
(Arnon and Stout, Plant Physiology. 14. 371, 
1939). Copper in certain minute concentrations 
is of the first importance in many cases. As ex- 
amples we may cite the well-known facts that the 
flagellate Eiiglena can live in a concentration of 
10~* M copper sulfate as long as food lasts, but 
dies in a few minutes when the concentration 
reaches 10"' M (Seybold, Biol. Zcbtralb., 47, 102, 
1927) ; that Amoeba proteus cannot survive when 
the concentration of CuCU rises as high as 
2 X 10-« M (Chalkley and Voegtlin, U. S. Pub. 
Health Rep.. 47, 535, 1932) ; that lysis of certain 
red blood cells by glycerol can be inhibited by an 
amount of copper insufficient to cover more than 
one-tenth of one per cent of their surfaces ; and 
so on. The fact that a fraction of a milligram of 
copper is just as necessary for the life cycle of a 
tomato plant as several hundred milligrams of 
potassium, for example, and the other facts cited, 
I would interpret as one more symptom of the 



THE COLLECTING NET 



August 30, 1941 ] 

fundamental superspecificity of biologically active 
structure systems. For the specificity of individ- 
ual native protein units, conjoined with specificity 
of interlinking to give the sui:)erspecificity of na- 
tive protein frameworks implies the existence of 
certain definite complements of positions capable 
of accommodating the metallic ions. While we are 
not yet in possession of complete information as 
to all the atomic environments into which ions 
such as zinc, copper, calcium, etc. can fit there is 
already sufficient indication in structure analyses 
to indicate that each ion type has its own individ- 
ual requirements, shown by the cation radius as- 
sociated with each cation coordination number. It 
would seem then that the stoichiometry of native 
proteins and ions, especially in relation to what is 
already known about atomic environments appro- 
propriate to the various ionic species would afford 
a direct experimental approach to the part played 
by metallic ions in physiological situations in cer- 
tain cases. Particularly significant in this connec- 
tion are the many facts from various fields indi- 
cating the part played laj' calcium ions, which may 
most usefully be studied in the light of structure 
analyses. These include the increase of the ag- 
glutination of sperm in the presence of calcium 
recorded by Loeb in 1914; the capacity of a torn 
sea urchin egg to mend itself in the presence of 
CuCL recorded by Chambers in 1924 (Am. J. 
Physiol., 72, 210, 1924) ; the studies on calcium 
caseinates by Philpot and Philpot in 1939 (Proc. 
Roy. Soc. London. 127B. 21, 1939) showing that 
particles of increasing weights are formed in solu- 
tions of increasing calcium concentrations. To 
my mind, all these suggest some specific inter- 
linking of R-groups of native proteins, modelled 
perhaps on the atomic environments characteristic 
of the ions in Ca(OH)i;, etc., a suggestion which 
could be exploited in detail in carefully designed 
experiments. The fact that calcium is known to 
be present in the nuclear membrane indeed 
prompts the suggestion that calcium ions (or 
some similar ions) may play an important part in 
holding together the native proteins which no 

INVERTEBRATE CLASS NOTES 

(Continued from page 184) 



191 



doubt constitute this and other biologically active 
membranes. In this case, a change in pH reduc- 
ing the ionization of the acidic R-groups riveted 
together by the calcium ions, would necessarily 
lead to a collapse or dissolution of the membrane 
in dividing cells. The subsequent formation dc 
novo of nuclear membranes in daughter cells 
would then require the accession of a definite 
though very small complement of calcium ions. 

In conclusion, I would record my belief that 
the present-day picture of the native protein is 
pregnant with meaning for all structure problems 
in physiology, because it offers hints as to direc- 
tions in which explanations of certain so far un- 
interpreted facts may be sought. I desire to put 
before those concerned with the design of experi- 
ments intended to throw light on the structure 
systems of living matter, the suggestion that it is 
the specific nature and behavior of individual na- 
tive protein units which is the central theme both 
for the understanding of the synthesis of proteins 
and for the understanding of the part played by 
metallic ions in living matter. It has already Ijeen 
shown that by means of these simple ideas both 
the high water content and flexibility of living 
matter and the immense specificity and stoichio- 
metric significance of living structure systems can 
be explained. Evidence is accumulating day-by- 
day that it is the intimate atomic structure of the 
protein molecule which dominates the scene. How- 
such a situation could possiljly be explained in 
terms of the curious hangover picture of polypep- 
tide chains of immense length I leave to others to 
explain. How suggestively, on the other hand, 
the new picture of the native protein fits in and 
comes to the rescue will, I believe, become in- 
creasingly clear when the perfectly definite indi- 
cations regarding its structure are made use of in 
the design of experiments on these essential con- 
stituents of living matter. 

(This article is based upon a seminar report pre- 
sented at the Marine Biological Laboratory on 
August 19.) 



— five to four ; the Inverts were victorious. 

Right in the middle of the game, fourteen of the 
girls strolled quietly to the supply house. A well- 
dressed young man approached and was about to 
enter the supply house when the girls graljljed 
him by the arm and marched to the Eel Pond, 
while the hurdy-gurdy man played "Who's Afraid 
of the Big Bad Wolf?" His adversaries allowed 
him to remove his outer garments, after which he 
meekly plunged in. A new M.B.L. record had 
been established : the girls had thrown a man into 
the pond. The fellow had taken a former dip 
into the Pond ungraciously and had taken the 
matter to the authorities. A meeting was called 
at which time he was to identify those responsible. 



It was at this point the Invert Amazons stepped 
in. To commemorate this deed and to express 
their gratitude, the "boys that sweep the lab and 
the boys that bring the 'fish' " arranged a fitting 
rally on the Brick Lab steps. With the inimitable 
Jasper Trinkaus presiding, there was much ban- 
ter, singing, and presentation of flowers. 

A strange figure looked in on the unusually 
hilarious group Tuesday evening, remarked: "My 
God ! this course isn't what it was when I took it 
forty-five years ago !" 

Saturday, August 30, is the last day of the 
course. We are especially grateful to the staff 
and crew ; this has been an unforgetable experi- 
ence. — Louise Gross and Bill Batchdov 



192 



THE COLLECTING NET 



[ Vol. XVI, No. 147 



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August 30, 1941 ] 



THE COLLECTING NET 



193 
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194 



THE COLLECTING NET 



[ Vol. XVI, No. 147 





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August 30, 1941 ] 



THE COLLECTING NET 



195 



SPENCER UNIVERSAL MICROSCOPE LAMP 




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196 



THE COLLECTING NET 



[ Vol. XVI, No. 147 




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