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Vol. XVI, No. 1

SATURDAY, JUNE 28, 1941

Annual Subseription, $2.00
Single Copies, 30 Cents.

THE SPRING CRUISES OF THE
ATLANTIS TO GEORGES BANK
Dean F. Bumpus
Woods Hole Oceanographic Institution

The Atlantis made four cruises to Georges
Bank this spring, one in March, one in April, and
two in May, in order to continue the studies of

the productivity of Georges

THE PHYSIOLOGY COURSE AT THE
M. B. L. IN 1941
Report of the Staff,
Physiology 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

Bank, that very productive
shoal water region extending
189 miles to the east of Cape
Cod. This series of cruises
curing the spring months when

MW. B. FE. Calender

THURSDAY, July 3, 1941 |

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

productive rate of plants and or group student research
animals is very great is a con- 8:00 P. M. along the lines of Cellular
tinuation of a program com- He 8 lke Mme nteouruien Respiration, Comparative
menced in the spring of 1940 Lecture: Physiology of Nervous Sys-

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 Wellesley College,

Mi George C. Whiteley, Jr, |
of the Hill School, and Mr.
Dean F. Bumpus of the Woods
Hole Oceanographic Institu-

| July 8.

“Current approaches to the Plant
Hormone Problem.” |

Dr. George S. Avery, Jr.,
Professor of Botany,
Connecticut College for Women,
New London, Connecticut |

The first weekly seminar of the |
season will be held on Tuesday, |

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 Membranes and Blood
Gas Studies. This will be
| conducted by the new mem-

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)

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

TABLE OF CONTENTS

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

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

The Spring Cruises of the “Atlantis” to
Georges Bank, Dean F. Bumpue..................0
Relation of Macronuclear Regeneration in

Paramecium aurelia to Macronuclear Struc-
ture, Amitosis and Genetic Determination,
Dr. T. M. Sonneborn
Notes on Eulima oleacea Embryology, George
M. Gray
National Education Association at Woods Hole

1
1

3

4
it

Scientific wAlpparabusy crests 8
Physiology Class Notes . . 10
Botany Class Notes ...... ¢ ial

Embryology Class Notes

Introducing Dr. Olga Janowitz 12
Ttem'ssofiinteres te tive erescieeseecsccceercers 3
Cold Spring Harbor Symposium .... 14
DinectoryahOne lO Alien eres a ne reaeoe er ecco 15

GHIOH SGOOM NI SHIYOLVYORVT TVOIDOTIOL AHYHL AHL FO NOILVOOT AHL SNIMOHS MOAIA TVINHVY NV

“SSRTV ‘DIOTNIG MON ‘POOM ‘WW PreMOoT, Aq YdeR1s0,04q

June 28, 1941 ]

THE COLLECTING

NET

|W

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 Paramecium 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 be given, depend upon the
discovery here reported of a new and unique
method of nuclear reorganization.

Paramecium 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;
but 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
absorbed 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, before 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 unabsorbed macronuclear
fragments in individuals that have in addition a

THE COLLECTING Nev 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,
It is published weekly for ten weeks between July 1 and September 15 from Woods

marine biological laboratories.

Hole, and is printed at The Darwin Press, New Bedford, Mass.

Mass. Single copies, 30c by mail; subscription, $2.00.

1938. It is devoted to the scientifie work at

Its editorial offices are situated in Woods Hole,

4 THE COLLECTING NET

[ Vot. 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 multiplication 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 be
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 subsequent 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 may
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 micronu-
clei, though not indispensible, nevertheless have
certain direct effects on cell characters. These
are first steps towards a solution of problems of
cytogenetics in Paramecium aurelia; 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

GrorGE 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.]

PREFACE

In August 1932 the author published in THE
Cottectinc 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 oleacea, of Kurtz and Stimpson, or

Melanella oleacea 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 (Thyone briareus), it is
easier to get them by collecting the sea cucum-

June 28, 1941 |

THE COLVECTING NET 5

250 Thyone and got seven Eulima, 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 Eulima 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 Eulima.
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, which 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 surrounding 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 observed 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, probably 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 bumped 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

6 THE (COLERCLING NED

[ VoLt. XVI, No. 138

individuals closely packed. No veligers yet
(GOSS NG INE.

At 6 P. M. one more capsule of eggs with the
six Eulima 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 Eulima. Beginning to show clearer individual
outlines. Still growing, but not veligers. The
Thyone feed was evidently an inspiration to the
Eulima.,

Twelve fresh Eulima were collectly yesterday
on Thyone, separated from the Thyone and placed
in a finger-bowl of clean sea water, but 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 —8 A. M. Veligers in first lot of eggs
still going strong but still in the doghouse. Seems
to be no indication of coming out. A few more
egg capsules were in the bowl with the same six
Eulima 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 Fulimna
(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 Thyone. 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 be that a good feed on the
Thyone 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 Thyone and three have already at-
tached to the Thyone. At 5:45 still three on the
Thyone, 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 capsule. Of the capsules from the six
Eultma, 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 Thyone 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 movy-
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 Thyone 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 Thyone, the other
six were sulking on the sides of the finger-bowl,
five were under water and one barely above the
surface.

July 22 — §:30 A. M. This A. M. four of the
ten last collected Eulima were on the Thyone.
The other six were mostly on the sides of the
bowl. But no eggs were found either on the
Thyone 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 Thyone had eviscerated. I threw out this
Thyone, washed the inside of bowl, put in another
Thyone 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
Thyone in different bowls had no egg capsules,
neither on the Thyone nor on the bowl. These
lots were: one lot of four from the catch of July

June 28, 1941 |

THE COLLECTING NET 7

8th, one lot of ten Eulima from the collecting of
July,17th. The veligers from the lot of six Eu-
lima 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
Eulima 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
Eulima 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 Eulima collected the 17th and
which had been on the Thyone and then taken off
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 Eulima have been laid for several days. On
July 31st six Eulima just collected were brought
to me and put in a separate dish. On August Ist
and 2nd more just collected Eulima were brought
to me and on August 2nd one more freshly col-
lected Eulima was given to me. These were all
placed in the same dish with those of July 31st,
and could not discover that any of them had laid.
On August 3rd two additional freshly collected
Eulima 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-
bowl from the others.
August 6. Two egg capsules in the bowl of
six Eulima. 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 Ist
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 Week)

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.

8 THE COLLECTING NET

[ Voc. XVI, No. 138

HANS DRIESCH, PHILOSOPHER AND SCIENTIST :

RALPH C. BUSSER

American Consul General at Leipzig, 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
October, 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 between 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-
phy of Organism” (1907/8), which was first
published in Great Britain, as it consisted of the
“Gifford Lectures” for which he received the
Gifford 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 professor extraordinarius.
In 1912 he published 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
position 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
Peking, 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
abscence 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
University 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 celebrated 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 Y)

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 of 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
Germany during recent years.

+ 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 Oz, 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 developed, 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
both 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
bottom 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 apparatus.

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 supplies 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
returned. Jn emergencies these may be recalled
for redistribution.

2. Glassware, including aquaria and museum
jars, be cleaned thoroughly before returning to
the Chemical or the Apparatus 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. XVI, 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 lab, with the remains of starfish, frogs and
dead dogfish hanging out of the waste baskets,
and the rest of the day’s rubbish ankle deep on
the floor. It is without a doubt the janitor’s
paradise.

The prevailing 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 beard, the centrifuge centrifuging and the
aspirator aspirating.

If Mr. Warburg were only here he certainly
would be proud of Dr. Fisher’s busy students.
They have run the Warburg apparatus almost to
the point of exhaustion, as observed by the mighty

screech it developed and the more than once
broken 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 lab, 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 staff 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, but also an electrician, a mechanic, a chem-
ist, a physicist, and, above all, an optimist.

—J. E. H.

BOTANY CLASS NOTES

Skepticism, instilled by certain members, 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 Chlamydomonas feverishly dashing across the
slide like army men heading for town on a day
off, and those who had never before met Chlamy-

domonas 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-
bert, 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 day
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.
Crest 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
particular 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.

EMBRYOLOGY

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 limbata 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-
dulus 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.

Friday being a special day for the botanists,
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: Tf 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.

—J. W. and C. S.

CLASS NOTES

Dr. Schotté 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 Hollister and Elizabeth Kirkpatrick

IZ THE COLLECLING NET

[ Vor. 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 238, 1938.

Introducing

Dr. Orca JANowi1tTz, 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 Vienna, Austria,
where she studied at the University of Vienna
and received her doctorate of philosophy in both
zoology and botany.

After she received her degree, she taught in a
Realgymnasium in Vienna, 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
principles of biology would be extremely useful
in providing a Weltanschauung 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.

Such lectures naturally brought 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, under a grant from the British
Federation of University Women, she conducted
research with Dr. H. Hardy at the Hull Univer-
sity College. He had devised apparatus to col-
lect plankton 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 conducting 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
spread 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. W. Ballard, Dr. C. Hyman, Warren
Healy, Dr. Lauren Gilman, Barbs Schraff, and
Mary Donovan.

While chatting with various 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, Mrs. 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. Wixzur has been appointed the
new hostess for the M.B.L. Club 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 ALM esi
June 28 . 7:04 = 7:20
June 29 7250) F809
June 30 8:37 S950
July on 1 9EZOMRSESS
July . 10:23 10256
July ) PLS SS)
July IZED
July LEG
July 2:12
July 3:08

. 12:59

1:56

. 2:54

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

ITEMS OF

Dr. Artuur K. Parpart, 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. Cart 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.

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 tlenson in
room 217 of the Brick Building, or Mrs. D. Mor-
ris as soon as possible.

The course in protozoology at the Marine Bio-
P gy at the
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 laboratory 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 hol1-
day.

INTEREST

Rurus H. Tuompson, 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 Permeability”. 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 Uni-
versity spoke on the “Carbohydrate Metabolism
of the Large Brown Algae.”

Mr. Epwarp 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. Chambers is a medical student
of New York University.

Mtss EpiruH Bittrnes, 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 the 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. Lancefield; Dr. T. K. Ruebush is Secretary-
Treasurer. Albert Stunkard continues as grounds-
keeper.

14 THE COLLECTING NET

[ Vor. 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
Laboratory 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 Laboratory. The aim is that
every important aspect of a given subject should
be 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.

Each day's program begins at 9:30 A. M.
When three papers are scheduled for the same
day, the third one will be read at 2:15 P. M.

STRUCTURE OF CHROMOSOMES AS REVEALED BY
OPTICAL METHODS

Wednesday, June 18th

Warmke, 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.

SALIVARY 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 chromosomes.

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.
viruses.

Mutation in Drosophila, bacteria and

PHYSICAL ASPECTS AND TOOLS
Friday, June 27th

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

Zworykin, V. K. Electron 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 of 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. Resumé and prospectives.

June 28, 1941 ]

Lap COLEECRING NED 15

DIRECTORY FOR 1941

Laboratories Residence

Botany Building.......... Bot Apartment
Brick Building... ...Br Dormitory .....
Lecture Hall...... L Drew House...
NiewneRoem in Fisheries Fisheries Residence ...... F

Laboratory.............0000 M ne
Old Main Building......OM Kahler
Rockefeller Bldg.....Rock Kidder .....

Supply Dept.................08 Ss

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.

McClung, 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.
Schotté, 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, K 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 COOLER CIING NET

[ Vor. 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.
Brey

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, I. 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
45. 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.
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.

Pub.

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. grad. phys. Toronto.

Grave, C. prof. zool. Washington (St. Louis). Br 327.

Groupé, 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 308.

Hamilton, Pauline G. grad. res. asst. zool. Pennsyl-
vania. Br 220.

Harris, D. L. instr. zool. Pennsylvania. Br 217-e.

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

18.

Harvey, E. N. prof. phys. Princeton. Br 116.

Harvey, 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. biophysies. 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 2038.

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 348.

Illick, 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.). 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., Il. 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
Phys.

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.
Med. Br 108.

Miller, J. A. instr. anat. Michigan. Br 228.

Milne, L. J. assoc. prof. biol. Randolph-Macon. Br
23, 1D) PANY

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).

Osborn, C. M. instr. anat. Ohio State. OM 46.

Osterhout, W. J. V. mem. Rockefeller Inst.
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. zocl. 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. [linois. 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
(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.

| Shelanski, L. grad. zool. Pennsylvania. OM Base. Dr

5

Pennsylvania

(New

Inst.

| 15.
Shelden, F. F. instr. phys. Ohio State. Br 111. K 24.

Sichel, Elsa K. head sci. dept. State Normal Sch.
(Johnson, Vt.). OM 4. K 8.

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

Yy 4,

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

Slifer, Eleanor H. asst. prof. zool. Iowa. Br 217-a.
D 203.

Sry J. A. instr. phys. & pharmacol. Chicago Med.

r 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
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.

penne Lois E. asst. prof. zool. Smith. Br 217-b.

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.

Le ee asst. prof. bot. Lingnan (China). Bot 1.

r 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.

(St.

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.
Kea 2:

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.). 5

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. grad. anat. Maryland Med. phys.
Bayard, Ellen B. U. of Connecticut. bot. H 8.
Bergstrom, 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 COLLECING NER

[ Vor. XVI, No. 138

burg, Pa.). emb. H 1.

Crumb, Cretyl Wellesley. emb. H 1.

Deyrup, Ingrith J. Barnard. phys.

Egan, R. W. res. asst. biol. Canisius (Buffalo, N. BYeS))s
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.
Ke Tie

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.

Karezmar, 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.
lel 7

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.
Val iff

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.

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

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

Rollason, H. D., Jr. grad. asst. biol. Williams. phys.
Dri.

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

Royle, 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, Ill.). 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, Betty 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. 8S.

Weisiger, J. R. fel. physiol. chem. Hopkins. Br 224.
1D a bila le

Apparatus and Technical Service

Boss, L. F. techn. Br 6.
Bridgman, Jane grad. biol. Harvard. 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

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.

Egloff, F. collector.

Gilbert, W. J. Michigan. bot. 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.

McInnis, J. mgr.

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

Parker, J. collector.

Poole, Margery bot. 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.
Gorokhoff, 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.

Schallek, 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.

Office 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

20 SOE ESS EE SSS 1 OSES

> OD SD) ) ED 0D (

0 O-SEE >

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Written by twenty leading specialists,
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June 28, 1941 | THE COLLECTING NET 21

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July 14th to 26th.

GOLD SEAL Cover Glasses Made in U. S. A.

WE ARE PROUD to be able to offer you careful laboratory testing and control...

cover glasses entirely made in the United Physically, the glass is practically free
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own workshop from imported glass. Now, is considered that the cost of domestic
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are proud of this accomplishment. please write for one on your letterhead.

American made GOLD SEAL Cover
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ADAMS ANGLE CENTRIFUGE

These centrifuges offer important advantages over
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See our Exhibit in the Old Lecture Hall,
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22 DH COLLECRING INET [ Vor. XVI, No. 138

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Credit for the leadership of Turtox microscope slides in American schools goes to the
highly trained individuals who make these slides in the Turtox Slide Laboratory. These men
and women have specialized in their particular fields and devote their entire efforts to just
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In addition to the usual mitosis material sueh as onion, Tradescantia, Ascaris, crayfish, Amby-
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June 28, 1941 |

i
:
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THE COLLECTING NET

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

Spencer has improved

STEREOSCOPIC MICROSCOPES

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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A booklet describing the complete line
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Sales Offices: New York, Chicago, San Francisco, Washington, Boston, Los Angeles,
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See exhibit of complete Spencer equipment in Canteen Building,
Bureau of Fisheries, June 30 to July 11.

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[ Vou. XVI, No. 138

Vol. XVI, No. 2

SATURDAY, JULY 5, 1941

Annual Subscription, $2.00
Single Copies, 80 Cents.

PROTOPLASMIC STREAMING
Dr. Doucras A. MARSLAND

Assistant Professor of Biology,
Washington Square College, N. Y. University

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

HOW FEATHERS ARE MADE
Dr. FRANK R. LILLIE
Professor of Embryology, Emeritus,
University of Chicago

I. Introduction

If this question could be fully answered within
the limits of present scientific principles and
methods we would have much

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

Seminar:

Shapiro.

Lecture:

RM, B. FE.

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

Physiological seminar in
which the following
tors will take part:

mian, Rita Guttman and Herbert

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

“The Integration of Neu-
rology and Psychology,” Dr. K.

| deeper insight into problems

of physiology, genetics and de-

velopment. But our knowl-
| edge is still very incomplete ;
| and of the little that we know
I shall be able to tell you only
a part of my own work, done

Calender

investiga- | originally with the aid of Dr.
A. J. Dzie- | Mary Juhn and recently of

Mr. 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

cyclic flow of protoplasm S. Lashley, Research Professor | of its growth and development.
which is characteristic of so of Neuropsychology, Harvard | By. plotting rates of axial
many plant cells, and the ebb University. growth with reference to the

and flow of pigment granules

location of simultaneous reac-

back and forth in the fine

branches which radiate out from the pigment cells
of the skin of many fishes,—all of these forms
of streaming are re- (Continued on page 31)

tions in the germ (isochrones )
it is possible to relate any definitive part of the
feather to its place and time of origin, and hence
to the conditions of determination when these are

TABLE OF

How Feathers are Made, Dr. Frank R. Lillie.... 25
Protoplasmic Streaming, Dr. Douglas A.

Ra SLAY Chemmecen sees tect ceevodcet eteeseesscessaciterencecessccceress 25

Notes on Eulima oleacea Embryology, George

Vibe Grrretiyau (COMMS)! eossstetecreceererececsecsctaeteesescoe carn cacs 29
Grafting of Limbs in Place of the Eye in
Amblystoma, Dr. Jean Piatt ....0..... ce eeeeeeee 32

CONTENTS
M.B.L. Department of Chemical Supplies and
SGlentifie App aratUSpeccc:c-ccc-ssccssssscetereacosessnceceses 33
Children’s School of Science ............. . 04
Items of Interest «0.0... 35
Physiology Class Notes ..... 36
Botany Class Notes ........... . 36
Embryology Class Notes ... 37
AUBUC! of MVWiOOdS) Elole) tasccesceseceotre-cetsscceseceeneee 38

‘punog paeAourA UO 4yNo Suedo aAOD OYJ, “PUNOASYOVG oY} UI USES eSnoYzYSI] 04} eau yoveq ey} uO oTUdId
jenuue a10y} oye} 0} AxOzeIOGe] 9Y} JO sossB[D SNolweA oY} 1OF ATewojsnd useq sey 4 SIveA IO

HAOD NITAVdIUVL GUVMOL DNIMOOT ‘NOHSOVN JO SGNVIdN WHHL

Jury 5, 1941 ]

Dt 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.

II. 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 by budding and
branching 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 barb 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. (II-
lustrated by 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 burst 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.

GHEE

The purpose of this lecture is not a general re-
view, but to present an account of experiments in
which old methods of experimental embryology
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

Operations on the Papilla

THe CoLiectinc NEevT 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.

marine biological laboratories.

Mass. Single copies, 30e by mail; subscription, $2.00.

It is devoted to the scientific work at

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,

28 THE COLLECTING NET

[ Vor. 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 breast follicle from which
its own papilla has been 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 papilla entirely free from all its
attachments in the follicle, and rotate it 180°
within the follicle, an upside-down feather results,
ie., 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, ventral 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 barbs 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 chimera 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”

Jury 5, 1941 ]

DAE COOLER CRING NED 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 barb 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, 1.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 apex 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
barbs. 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 differentiation. 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 Thyone having gone bad
and no Eulima eggs that I could discover, I threw
out the Thyone, gave the Eulima 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-
lima 1 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
3lst 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 Thyonc, 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 3lst 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

sO

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 3lst. That day I changed the water more
thoroughly, leaving them in the same jar.

On September Ist 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
Eulima 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 Ist 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 COLLECTING NET

[ Vor. XVI, No. 139

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.

Jury 5, 1941 |

THE COLLECTING NET 31

March S8. Looked carefully this morning and
decided two more egg capsules had been laid since
the sixth. There are just 16 Eulima in the bowl
with the eggs, proving that one snail at least
must have laid twice. 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 Eulima might lay at this season as some of
the Nudibranchs do. But I have my doubts about
Eulima doing so.

March 10. This morning the egg 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 Eulima 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 Eulima 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).

After this there seems to 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 Eulima 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 Fu-
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 Eulima, and five
live snails. I cleaned the dish, leaving the eggs
and took out the dead snails’ shells and put back
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 Eulima was brought to me on June 25th. 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 Ist 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 breeding 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
Ibs. 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-

32

THE COLERCRING NEG

[ Vot. 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
objective would not be suitable for viewing tis-
sues through so thick an intervening layer of
glass, a special objective, with a working distance

GRAFTING OF LIMBS IN PLACE

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 on’ 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 Structure” at the meetings of the

American Association for the Advancement of
Science, December 30, 1940.)

OF THE EYE IN AMBLYSTOMA

Dr. JEAN Pratt

Departments of Anatomy, University of Vermont, Burlington, Vermont and College of Physicians
and Surgeons, Columbia University, New York City

When a forelimb rudiment is grafted to the
head region of salamander embryos, the resultant
limb 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 been transplanted in place of an eye.

Nicholas (29, ’30, ’33) made homoplastic and
heteroplastic forelimb grafts to the eye region in
Amblystoma. 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-
hibited individuated movements of the various
parts. 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 down 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 experiment 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 (736) 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
possibly 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 larvae 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

Jury 5, 1941 }

THE COLLECTING

NET 33

thought that contraction of these muscles would
account for the feeble, stereotyped movements ob-
served 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
branches 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 difficult to, re-

move all the eye muscles in such an operation and’

that the coordinate movement of the limb with
the opposite eye is caused by the remaining eye
muscles gaining an insertion onto the base of the
transplant. If this is so, then the coordination of
limb 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 Room. 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./1 :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

[ Vor, XVI, Nowmis9

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.

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:
stitution, Cold Spring Harbor).
in Plants.”

A. F. Blakeslee (Carnegie In-
“Growth Patterns

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). “Heterogonic Growth.” The ontogene-
tic and phylogenetic significance of differential rates
of growth.

Afternoon session: L. C. Dunn (Columbia Uni-
versity). ‘Abnormal Growth Patterns, with 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. MacHaffe 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 Study 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-
lating season ahead. —Mrs. S. McMurtrie.

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 AM.) PaMe
July 5 SH2ZESO SIG
July 6 150m ZellZ
July 7 2:54 3:08
July 8 3:48 4:00
A Cline heacetete weaceene Rye AbEDS)
July 10 . 5:28 5:40
italy lb ee cee Gps} |sst0)

—— EEE

Jury 5, 1941 ]

DE COLLE GRNG

NET 35

ITEMS OF

Dr. EuGcene F. DuBots, professor of medicine
at the Cornell University College of Medicine, has
been 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. Donatp 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 1s 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. Ropert A. Gorrin and his staff 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 Fisheries was 3,401. From July 1,
1940 to June 30, 1941, 25,099 persons wrote their
hames in the visitors’ book. In general, Mr.
Goffin finds that about one in four persons who
come to the aquarium register there.

INTEREST

At 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 NAveEz, 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
Mal.

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. RuesusH, 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. Br
Tale

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 3824.

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.

Trager, W. assoc. Rockefeller Inst. (Princeton). Br
208.

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

36 THE COLLECTING NET

[ Vor. XVI, No. 139

PHYSIOLOGY CLASS NOTES

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
Warburg 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 subject 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 basement 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 arbacia, 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.”

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
through 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
being made by the committee, Bob Harrison and
the Black Phantom (his car). —J. E. H.

BOTANY CLASS NOTES

Sometimes I have to wonder why
The artist’s life I did deny,

For plants, I find, are not elating
And skunky Chara’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—all 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

Juty 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-jeaned
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.

EMBRYOLOGY

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 recetved. 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
phylum Chordata, to which Homo sapiens be-
longs.

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. —J.W.

CLASS NOTES

The week ended with a lecture by Dr. Schotté
(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.

THE, COLEECLING NET

The A. B. C. of Woods Hole for 1941

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

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

10:30 to 5:15

Station Open 6:00 to 8:00
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
8:00 to 9:00

Sundays
10:00 to 12:00
4:00 to 6:00

Holidays
9:00 to 11:00

4:00 to 6:00

POST OFFICE

LIBRARY HOURS

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.

*All trains stop at Falmouth.

TRAIN SCHEDULE*

{Discontinued after August 31.

Ex. Sat.

Weekdays Weekdays & ane Weekdays Sundays Sundays
Woods Hole 7:06 10:15 12:55 5:45 6:00 7:55
Boston 9:15 12:35 2:52 7:57 8:10 9:55

Ex. Sat.

Weekdays Sundays Saturdays] Weekdays & Sun. Weekdays
Boston 8:20 8:35 12:25 1:05 4:00 5:00
Woods Hole 10:45 10:45 2:30 3:25 5:59 7:10

Fri., Sat.,

Daily Weekdayst Sun.£_ Fridays!
2:30 or 7:30 é ene
3:50 7:15 8:45 9:30
4:50 peers ae 10:15
SAcrehe 8:00 9:30

7:00 aioWtehce 12:15
Daily Weekdayst Daily Sundays§
2:30 Soin 4:45 Pens
4:30 eer 6:45 9:00
5:30 6:45 7:30 9:45
6:45 Sate 8:45

£Runs 15 min. later on Sunday, daily after
August 31.
$Also runs Labor Day.

BOAT SCHEDULE*

Leaves Daily Daily Weekdays
New Bedford 7:00 9:30 2:00
Woods Hole 8:30 10:50 3:15
Oak Bluffs 9:20 11:40 4:00
Vineyard Haven F Bees 4:30
Nantucket (due) 11:35 2:00

Leaves Daily Daily Sundays{§
Nantucket mpeen 6:45 2:00
Vineyard Haven 6:10 Bet shiefisys
Oak Bluffs oe 9:00 4:00
Woods Hole 6:55 10:00 5:00
New B’df’d (due) 8:15 11:15 aan} c
*Schedule effective to Sept. 6, incl.
{Discontinued after Aug. 30.
£Does not run Labor Day.

[ Vor. XVI, No. 139

Jury 5, 1941 | THE COLLECTING NET 39

5000 R. P. M. on D. C.

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40 THE COLEECTIING NED [| Vor. XVI, No. 139

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Jury 5, 1941 |

THE COLLECTING NET

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41

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42 THE COLLECTING NET

[ Vot. XVI, No. 139

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44

THE -COLLECTING NET

HROUGH the cold dank dusk a watcher

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[ VoL. XVI, No. 139

Vol. XVI, No. 3

SATURDAY, JULY 12, 1941

Subscription, $2.00
30 Cents.

Annual
Single Copies,

THE SOURCE OF PANCREATIC JUICE
BICARBONATE

Dr. Eric G. BALL
Associate Professor of Biochemistry,

CURRENT APPROACHES TO THE PLANT
HORMONE PROBLEM

Dr. GeorGE S. AVERY, JR.
Professor of Botany,

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- Hone the Atlantic Palolo
carbonate of the blood flowing | py Paul S. Galtsoff: “Accumula-

through the gland. The other
is the CO 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 COs 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
Such a relation can be
interpreted to mean that (Continued on page 55)

the bicarbonate content.

duced in the cells

MM. BD. £. Calendar

TUESDAY, July 15, 8:00 P. M.
Seminar: Dr. L. B. Clark: “A
Suggested Mechanism by which
the Moon Influences Reproduc-

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.”

Connecticut College

3y definition, whether in animals or plants,
hormones are chemical substances normally prd=

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 regulate 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, JY. .....cc.00 45

The Source of Pancreatic Juice Bicarbonate,

TABLE OF CONTENTS

IDs! IDhave, (Ces) 82) Ie eee er reread sore reerotecy eae 45
Physiology Class Notes
The Permeability and the Lipid Content of the H
Erythrocytes in Experimental Anemia, Dr. Botany, Class) Notes
ANsapobEE Dl, IDVATETTIWIES TY GaccoocnoocoscecncooeccnecooOECEEoDG 50

Symposia at the University of Chicago............
Introducing Dr. Cheng-Kwei Tseng

Items of Interest ...

Embryology Class Notes ...ccccccccceccsecstccssseesseees 55

“yJo] eular}xe oY} Je USES oq Ud o4BISe SesseqreM\ “Iq JO diy
ay} {yuL0g eouezueg SI punossyorq ey} UL puel Fo uogqit oy, ‘spuyls] Wey pues JOO SjIAeq ‘ould :o1e punorsyoeq oY} 04 punoide.10j
ay} wory Sulpeet SpULIST oy], “‘qySId oy} UO BADD Ueg desyg YIM punoidse10f; oy} UL Stvedde pues] JossewueUON JO pus YyNOS oY,

NUAMLAD SGNVISI AHL GNV ‘LNIOd AONVZNGd “LASSHANVNON AO MATA 'TVINAV

Cnn aaa

Jury 12, 1941 |

THE COLLECTING NET 47

cussed hormones in higher plants is vitamin By,
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 im 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 effects, 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 e¢ 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 constant 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 COz 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 fairly, 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.

marine biological laboratories.

Mass. Single copies, 30e by mail; subscription, $2.00.

It is devoted to the scientific work at

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,

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, 1.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 Avena 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. Avena 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 being 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 Avena method. Corn endosperm extracted
by this method gives yields as high as the equi-
valent 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-
portant, 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 may 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, ¢.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

Juy 12, 1941 ]

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.

Synthetic 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 e¢ 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, 1.e., that
once started they continuously secrete such sub-

THE COLLECTING NET 49

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 * WN. langsdorfii 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 growth.
Mention of mobilization effects attributable to
hormones has just been made, but it seems un-
likely that the primary effect 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 im 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 preliminary 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

[ Vor. 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 living tissues, more on
inhibiting substances 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
“erowth 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

Bodansky, (J. Biol. Chem., 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 effect 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,

ory 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.
After the 9th day the new cells began to assume
normal size slowly. Comparisons of frequency
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 im vitro has no effect
upon the permeability of the erythrocyte, 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 m 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 permeable
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
phenylhydrazine 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)

92 THE COLLECTING NET

[ Vot. XVI, 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.

Introducing

Dr. CHENG-KWEI TSENG, Assistant Professor of
Botany and Acting Curator of Herbarium, Ling-
nan University, China; University Fellow in Bot-
any, University 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 part 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 United States
to work at the University of Michigan under Dr.
Wim. 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, Wrangelia, Griffithsia, Codium and Chae-
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.

Bedian, D. asst. prof. anat. Western Reserve Med.
ib.

Brown, D. E. S. prof. phys. New York. Br 310.

Calkins, G. N. prof. proto. Columbia. Br 3381.

Chambers, E. New York Med. Br 348.

Chambers, R. prof. biol. New York. Br 328.

Duncan, G. W. fel. surg. Hopkins. Br 328.

Genevieve, (Sister) Mary grad. biol. Villanova. Rock

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

(New

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
July
July

P.M.
LPI
8:10

A.M.

7 :00

7:47

8:35

At eee OEZ3
pte ee ORS
aiken. JhILOY
ee ESS:

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

Jury 12, 1941 ]

THE COLLECTING NET 53

ITEMS OF

Dr. AND Mrs. CHARLES PACKARD will be at
home to members of the Laboratory on Sunday,
July 13, and on the two following Sundays, from
four-thirty to six o’clock.

Dr. Rogert GEORGE BALLENTINE was granted
a National Research Council Fellowship for the
year 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. Marcery MILNE has been appointed as-
sistant professor of biology in the Richmond Di-
vision of the College of William and Mary.

Dr. Dororuy 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.
Working 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. Eucene F. DuBois has been appointed
professor of physiology at Cornell University
Medical College, and head of the department of
physiology and biophysics, not biochemistry and
physiology as previously reported.

Dr. Joun H. Norturop, 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.
EE. G. Conklin lectured before the embryology
class today on the mapping of eggs.

Dr. Atma 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.’

INTEREST

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

Miss Murret Vorer, 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. Davin J. Braprey, 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. Rosperts RuGu 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. Ditton, 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 Mufioz); 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 18lst 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 NED

PHYSIOLOGY CLASS NOTES

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
difficult 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 butter 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-breaking with the aid of the
Topper series as reference.

Just a word about our favorite delicacy—amicro-
manipulation. At the beginning of the week we
handled the needles and pipettes as if we had re-
ceived our early 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 thereby eliminated. There also seems to
be an advantage because advice is available from

those who have gone before and know what not
to do. Thusly have the new micro-manipulators
learned that dirty needles and clogged pipettes
cannot be cleaned with a handkerchief, while the
new Van Slykers are still wondering about the
best 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 Wednesday.

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 offices of an
ex-officio physiology instructor who managed to
find time, despite the exigencies of his embryol-
ogy course, to instruct us in all phases of physiol-
ogy. We thank him. If, JB, Jal.

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-efficient Nancy Bull.

Upon docking, their first experience was an in-
side view of Maria Mitchell’s house, where the
telescope with 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 fauwa-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
slowly towed through the Gutter amid cheers and
catealls from the Nantucket. Unanimously pro-
claimed the best trip yet, this excursion listed only

[ Vot. XVI, No. 1407

Jury 12, 1941 |

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

THE COLLECTING NET 55

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
stop to appreciate a pair of deer that approached
them fairly closely before gracefully bounding off
again. Some of the city critters hud 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.

EMBRYOLOGY CLASS NOTES

Acadenmuc Life:

The past week has been spent on Echinoderms
with Dr. Schotté, who dwelt at first on the illu-
sive orange ring of the eggs of Paracentrotus. 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. Schotté 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. Schotté 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

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 skirmish 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. Onde

THE SOURCE OF PANCREATIC JUICE BICARBONATE
(Continued from page 45)

when the gland works hardest it produces the
most COs 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-

lyzes the reaction CO, + H2,O = H2CO3. Now
if the metabolic CO. 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 COLEECTING NED

[ Vor. XVI, No. 140

carbonic anhydrase in the pancreas may be looked
upon as an indication that this enzyme is needed
there in order to speed up the hydration of meta-
bolic CO» 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 observed 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 im vitro.
These results therefore indicated that carbonic an-
hydrase had nothing to do with production of pan-
creatic juice and suggested that the metabolic
COs 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 (C13). 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
radioactivity 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 bicarbonate 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 CO, 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-
periments 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 H2O 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 PANCREATIC
JUICE.

All values expressed in terms of mM/Kg HO.

Pancreatic Juice

Blood Pancreatic =
Plasma Tissue Slow Rapid
Secretion Secretion
Na 155: 158. 158.
k 5.4 6.0 6.0
Cl 120. TAL 108. 30.
HCOs 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
pancreatic 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.)

4
|
|

Jury 12, 1941 |

THE COLLECTING NET

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Again, Dr. Hunter presents a timely
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[ Vor. XVI, No. 140

Jury 12, 1941 ]

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59

60

THE COLLECTING NET

[ Vor. XVI, No. 140

Edward Bausch....

HILE 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

. Microscope Maker

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
that no better optical instruments can be had
than those bearing the Bausch & Lomb Trademark.

BAUSCH & LOMB

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ESTABLISHED 1853

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FOR NATIONAL DEFENSE, EDUCATION, RESEARCH, INDUSTRY AND EYESIGHT CORRECTION.

Vol. XVI, No. 4 SATURDAY,

JULY 19, 1941

Annual Subscription, $2.00
Single Copies, 30 Cents.

MAGNETIC MEASUREMENTS ON COM-
POUNDS OF BIOLOGICAL SIGNIFICANCE

Dr. L. MicHAELtIs
Member, Rockefeller Institute for Medical
Research

Diamagnetism and Paramagnetism are analo-
gous to induced electric dipoles and permanent
electric dipoles. In magnetism, there is no phe-
nomenon analogous to a free

THE MAPPING OF EGGS
A LECTURE TO THE EMBRYOLOGY CLASS

Dr. E. G. CONKLIN
Emeritus Professor of Biology,
Princeton University

I feel like apologizing for appearing before an
audience of people who are doing so much 1m-
portant work in this day and age, because I have
to report on things which were

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- Dr.

MM. B. E. Calendar

TUESDAY, July 22, 8:00 P. M.
Seminar: Dr. G.
Melanophore System of Teleosts.”

R. L. Watterson:
pects of Pigment Deposition in

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

H. Parker: “The

“Some As-

ments are always very small, entinss Coun Of Glide isn same terms or under the same
always involving repulsion by bryos.” topic.
an external magnet pole; and Dr. H. L. Hamilton: “The Influ- I was once introduced as

paramagnetic moments, if they

exist at all, are always rela- culation
tively large and cause attrac- DRUM oeae at RAK:

tion by a magnet pole. All
matter is diamagnetic, but in
certain chemical compounds, |
paramagnetism is superim-
posed on diamagnetism, and
being much larger, makes the
latter almost negligible. It may

ence of Hormones on the Differ-
of Melanophores in

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 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 marvels and wonders of
development.

I am therefore to give you

“The Distri-

be added that the phenomenon
of ferromagnetism exhibited by a few metals,
such as iron or nickel, practically only in the free
metallic state, is never (Continued on page 72)

something of a historical re-
view of the backgrounds of modern embryology.
The marvel of development never grows old or
stale. In a single night an ascidian egg will be-

TABLE OF

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

CONTENTS

Metabolism and Fertilization in the Starfish

Egg, Dr. H. Shapiro
Book Reviews
Editorial Page
Items of Interest
Physiology Class Notes
Botany Class Notes |
Embryology Class Notes ........cc:ccccccsssssssssssesseceees Ee ||
Map of Woods Hole

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.

ony 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 Molgula, 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 baby. 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
peanut, 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-
port 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 CoLLEctTING 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 published weekly for ten weeks between July 1 and September 15 from Woods

marine biological laboratories.

Hole, and is printed at The Darwin Press, New Bedford, Mass.

Mass. Single copies, 30e by mail; subscription, $2.00.

It is devoted to the scientific work at

Its editorial offices are situated in Woods Hole,

64 THE COLLECTING NET

[ VoL. XVI, No. 141

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

Then a young medical student, Gaspar Fred-
erick Wolff, published his doctoral thesis in 1789,
entitled “Theoria Generationis”. He made an
actual study of the hen’s egg and of certain plant
seeds and found that in the earlier stages there
was no little animal or plant in the egg, 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 formativus 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. Whitman showed the absurdity of this. He
asked how is organization going to be “cooked
into the egg” which lacks it? What nisus 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
which 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

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,
Pfluger, 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 Pfltiger, 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
Jedes.’ 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, entelechy. 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

Jury 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.
Why 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 entelechy. 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.

(Concluded 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. Heterophyes heterophyes,
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 Metagonimus 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 conditions
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 littorea, 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

[ Vor. 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 examination of
the surface shows few parasites 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 villous 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
parasites 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
very few eggs could be found and the feces were
negative at the end of 6 months. After resistance
had been established in dogs, the feeding of large
numbers of metacercariae produced no visible ill
effects and very 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
because 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

Jury 19, 1941 ]

THE COLLECTING NET 67

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 dumerlu. 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-

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
lunar 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 Chaetopterus, (J. Gen. Physiol.,16,
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

[ Vou. 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 punctulata, 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., 67, 468, 1931)
of the egg of the starfish Asterias forbesiu, 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 examines the
original data, it is noted that relatively low per-
centages of cleavage 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 June, 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.,
Sabellaria alveolata, Chaetopterus variopedatus,
Arbacia punctulata and Asterias forbesi, it ap-
pears 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 oxidative enzyme system on fertilization. For
Arbacia punctulata, this has been further demon-
strated by various 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-
portance, 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. Ina 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
many 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,

Jury 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. Schroeder

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 be found in any work
and then endeavoring to maintain the case that
they are major faults. The soundest test is prob-
ably to inquire whether the volume still justifies
its title.

Biology is a vast subject defined in the first
paragraph 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 biology. 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 co6rdination
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 foundation
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

[ Vor. 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 238, 1938.

M.B.L. CLUB

The Annual Meeting of the M.B.L. Club will
be held at the Clubhouse 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 used 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 part 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
FE. Hill, T. H. Bullock and J. Hutchens.

—Sears Crowell

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.

Kovac 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 RK. 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
2% ton 6 X 6 “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. Your 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 Ze
July 20 1:30) aes
July 21 . eft 2:20 eZesil
WOU ZZ ccastonacse 3:05) (/Sil5
July 23 . 3:49 4:00
uly 245 eee 4:30 4:43

Ss ———__

Jury 19, 1941 ]

THE COLLECTING NET 7\

ITEMS OF

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. Cart 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. Proves 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. RANnpoLtPpH 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 (1937-1938) at the University of
Wisconsin. He is well known for his work on
Dauermodifikationen 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.

INTEREST

Miss ELEANOR BLEVINS is being married today
to Dr. Donald J. Zinn, who has worked at the
Laboratory 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. Paut 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 MicwHaettis left Woods Hole with
his family for northern New England yesterday.

Miss Louise °E. BoypEN, 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.

THE COLLECTING NET

[ Vou. XVI, No. 141

ITEMS OF

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 Walter 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 Dohrn sailed for a routine
trip under the supervision of Mr. F. Fuglister.

INTEREST

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 dyestuffs such as methylene blue, pyo-
cyanine, the yellow respiration ferment, has been
previously demonstrated by a _ potentiometric
method, surveyed 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 paramagnetic, 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 paramagnetism 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.)

Jury 19, 1941 ]

THE COLLECTING NET 73

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 Limulus 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
communiqué 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 minima 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 rubrum 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. Lucl 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 plumosa 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

[ Vor. 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, but 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

EMBRYOLOGY
Academic Life:

On Tuesday, July 8, Dr. Schotté 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 SO, ion has in the polarity
of the egg. We are sorry that this course will
no longer have the privilege of Dr. Schotté’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 Nauplius 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

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
Cystoclonium—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. —J. W. and C. S.

CLASS NOTES

Dr. Conklin, who gave an introduction on the
argument of Preformative vs. Epigenetic Devel-
opment and concluded with a brilliant discussion
on “The Mapping 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 that the laboratory
be continued at sunset. —P. and K.

75

THE COLLECTING NET

Jury 19, 1941 }

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THE COLLECTING NET [ Vor. XVI, No. 141

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THE COLLECTING NET [ Vout. XVI, No. 141

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THE COLLECTING NET

[ Vor. XVI, No. 141

The Strange Case of The Invisible Evidence

De had struck in the night.

A fleck of copper on the suspect’s knife was
the only clue. But with this trifling bit of evidence
alone, criminologists using the spectrograph were
able to prove that the knife had cut copper. By
the percentage of constituents and impurities pres-
ent, they identified that fleck as having come from
that specific telephone wire. The case was solyed—
a murderer convicted.

Dramatic as has been the record of spectrog-
raphy in criminology, such spectacular feats are
dimmed by the everyday accomplishments of
spectrographers working in science and industry.

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develop and.control the metal alloys now so vital

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Vol. XVI, No. 5 SATURDAY, JULY 26, 1941 ae Chae eae

STORAGE AND DISTRIBUTION OF MAN-
GANESE IN OSTREA VIRGINICA
Dr. Paut S. GALTSOFF
U. S. Fishery Biological Laboratory

THE ORGANIZATION OF THE MELANO-
PHORE SYSTEM IN BONY FISHES
Dr. GEorGE HowArRD PARKER
Professor of Zoology, Emeritus

Harvard University

The common eastern catfish, Ameturus nebulo-
sus, 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 above. 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.
light that falls on the white background below the

The tissues
relatively large

Woods Hole, Mass.

of marine Lamellibranchs
quantities of heavy metals.

store

Fe,

Cu, Mn, Zn and sometimes Pb were found in

their bodies and it has been

M. B. LZ. Calendar

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:
Radiation on Regeneration
Nais paraguayensis.”

Dr. A. M. Chase: “Effect of Azide
on Cypridina Luciferin.”

“Effect of X-
in

FRIDAY, August 1, 8:00 P. M.

Lecture: Dr. Ernst Mayr, ‘“Specia-
tion in Birds.”

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-

Of the

or location, oy

buted to racial differences, age,
sters of known age and_ origin

were planted on the experimental bottom in Long

fish and is reflected (Continued on page 89) Island Sound near Milford, Connecticut. The
TABLE OF CONTENTS

Storage and Distribution of Manganese in The Distribution and Development of the Me-

- Catt 2 (@aivieoe. 81 lanophore Hormone in the Pituitary of the
Cee inicnel Es Paul iGaltso Chick (De AEreRabn' c 5 cote ee 87
The Organization of the Melanophore System Miditonial Rag ewicccctcccce eee . 90
in Bony Fishes, Dr. G. H. Parkev...............0 Sil Marae Ge Ibert 1. 91
Th * E -, EB. G. Conklin.......... 83 Physiology Class Notes ............... > CF
e Mapping of Eggs, Dr. E ois ee Botany Class Notes ..........c0:cccsseesees . 92
The Rectifying Property of the Giant Axon of Embryology Class Notes .....cc. . 93
the Squid, Drs. R. Guttman and K. S. Cole.. 86 Supplementary Directory for 1941 .......0.0000.. 94

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“M WOqGoY ‘209 “M Yodteperg “ap ‘suing paempy “f¢ “TYsty “O Yyyouuey “Aq ‘2301 "Y uyor ‘WIN|[quesoy ouedny ‘sqoovr your :MOI yorg
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IP6L ‘AUOLVUOAVI TVOIDOIOIN ANIUVW AHL LV ASHNOD AVO'TIOISAHd AHL dO SLNAGALS GNV dHiVLS

Jury 26, 1941 ]

3000 bushels of oysters planted on this area rep-
resented a homogenous population from which for
a period of about 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. p.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-

THE COLLECTING NET 83

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 7.3 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 8.9 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 Mn is stored, it is reasonable to con-
clude that higli content 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 (J. 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 ieport 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 Keimbezirke, 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 Rabl studied the embryology of
Planorbis 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, 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 published weekly for ten weeks between July 1 and September 15 from Woods

marine biological laboratories.

Hole, and is printed at The Darwin Press, New Bedford, Mass.

Mass. Single copies, 30e by mail; subscription, $2.00.

It is devoted to the scientific work at

Its editorial offices are situated in Woods Hole,

84+ THE COLLECTING NET

[ Vor. 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 young French student, Louis Chab-
ry, studied the ascidian egg, following the meth-
ods of Driesch. He found that one of the first
two blastomeres 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 Roux stated that the frog’s
egg from the four-celled stage on is a “mosaic
work”, each of these portions 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,
“Amphioxus 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 Crepidula,
but had never met Wilson and knew nothing of
his work on Nereis. Wilson heard of what I was
doing and one Sunday morning he came over to
see my drawings of Crepidula 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 blas-
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.
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 shows 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 Amphioxus, 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 by 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 egg,
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 was 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 cytoplasm 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

So the question arose,

Jury 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 what is happen-
ing in the egg to recognize some of the steps 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 Cumingia, 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 membra
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

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., Linerges)
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 axis 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-

THE RECTIFYING PROPERTY OF

[ Vor. XVI, No. 142

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 GIANT AXON OF THE SQUID

Drs. Rita GUTTMAN AND KENNETH S. COLE
College of Physicians and Surgeons, Columbia University

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

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.

E

|

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-

|

Jury 26, 1941 ]

THE COLLECTING NET 87

trodes 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 liv-
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 effect 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. RAHN

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

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

88 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
pars 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
separate 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
pars anterior is entirely responsible for hormone
production 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 differentiation 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-
nently 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 A(nolis) U(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 y
of tissue
5 present
7 0.2 —
9 0.8 —
12 5.0 9
15 11.0 13
21 (hatching) 90.0 60
49 166.0 56
adult 909.0 Wa

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

Jury 26, 1941 |

THE COLLECTING NET 89

seen that the concentration of the hormone per
unit mass does not increase after hatching. All
apparent 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 problem 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 Biclogical 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 shown 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-pituitary 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
be 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 IMENS, COMLIRCMUNG, INI

[ Vor. 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 28, 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 by
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 costume, 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 program 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. Roserr CusHMAN Murpuy, 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 PNSW I 1E% Wile
utlve 25: een eee S427
July 26 Hey sil
July 27 640 6:58
July 28 7:25 7:48
July 29 8:13 8:40
July 30 9:04 9:36
July 31 2 9258 OES6
August 1 LORS l38

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

|

Jury 26, 1941 ]

THE COLLECTING NET 91

ITEMS OF

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. Orto Graser, 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. Davin R. Gopparp, assistant professor of
botany at the University of Rochester, and in-
structor in the Botany course at the Marine Bio-
logical Laboratory, has been promoted to an asso-
ciate professorship at the University.

Dr. James D. Harpy, 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. Buttock 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. Georce E. CoGHiLt, 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.

INTEREST

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. Herpert 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. ANpERSON, business manager of Bio-
logical Abstracts, visited Woods Hole for two
days early this week.

Dr. AND Mrs. P. B. ARMstTRONG 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

Wiss; (COMMA CIMUNE, IN1B AT

[ 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. Barroén’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 even 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 officially 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 like to have
some fun, i
Just open all the stops and cocks and let it wildly

TUN ees
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 drugstore 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,
vim,

Not dashing from the lab each day to follow some
fool whim.

without that sparkling

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—

Jury 26, 1941 ]

THE COLLECTING NET

93

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.

EMBRYOLOGY

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 embryology 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

Picture Augusta without the Victoria,

Harriette, Antonia, and Leuchs in there, too;

Not puffing her corncob each nite after tea,

Wewune hair that’s unkempt and expression that’s
ue.

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. ‘
=f WV wine! (Oo Se

CLASS NOTES

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 begun 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 ways. —E. R.

THE ORGANIZATION OF THE MELANOPHORE SYSTEM IN BONY FISHES
(Continued from page 89)

coal-black. The blood 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 innervation 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.)

94 THE COLLECTING NET

[ Vor. 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. New York Med. Br 343.

Chambers, R. prof. biol. New York. Br 328.

Clark, Eleanor L. assoc. anat. Pennsylvania Med. Br
ible

Duncan, G. W. fel. surg. Hopkins. Br 328.

Erlanger, 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., III 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 3438.

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 318.

Sturtevant, A. H. prof. biol. California Tech. Br 126.

preeer, W. assoc. Rockefeller Inst. (Princeton). Br

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.

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. Wesleyan. 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. Radcliffe. 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.

Humn, D. G. grad. zool. Yale. D 311.

Kielich, E. R. asst. biol. Canisius (Buffalo, N. Y.).
Dryas

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. Harvard. 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.

Jury 26, 1941 |

THE COLLECTING NET

33 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-
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# manufacturer.

53 Due to our desire to continue our con-
3 tacts with those who are doing research, we
zz recently set up a new shop to be devoted
#% ©entirely to the designing and making of
#% = experimental apparatus.

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In this manner, we will be able to con-
tinue the manufacture of Warburg Ap-
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Mr. John Linden, who is in charge of
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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-

ments may not be used to advantage.

Cambridge products include Galvanometers,

Electrometers, Fluxmeters, Vibrographs,

Geophysical Seismographs, pH Meters and

Recorders, Gas Analyzers and many other

mechanical and electrical instruments of Bp.
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Write for literature of instruments of use CAMBRIDGE POT GALVANOMETER

to you. This galvanometer is an inexpensive instru-
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HUMAN SKULLS

For several years 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 150

skulls, most of which were of unusually fine quality. These are now
offered at prices of from $74.00 to $35.00 depending upon the com-
pleteness of the dentition and general perfection of the skull.

It will pay any school to take advantage of this offer in providing
skulls for demonstration purposes. We will gladly send teachers
several for inspection with return privileges.

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761-763 East Sixty-Ninth Place, Chicago i
The Sign of the Turtox Pledges Absolute Satisfaction |

Jury 26, 1941 | THE COLLECTING NET 97

Porret

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sodium light.
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magnifier.

Tripartite Contributors

identity field.

CZ “<a ‘\
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These, and other slide boxes and trays given
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THES The gies. CLAY-ADAM Inc.. New York, N. Y.

98 THE COLLECTING NET

[ Vor. XVI, No. 142

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Use these Forceps for han-
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Order a few—you will use
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Jury 26, 1941 |

THE COLLECTING NET

99

@ Spencer announces a Direct Result
Colorimeter which presents many im-
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day instruments.

1. Direct reading of the percentage of
the unknown.

2. Inclined eyepiece for comfort.

3. Simple, accurate adjustment of arti-
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4. Tilted plungers which dissipate air
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5. Cups which are easily taken apart for
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A New Spencer
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6. Reverse position of instrument affords
convenience in placing or removing
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7. A beautiful, easy to clean, modern
design that combines strength and
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Your nearest Spencer sales office
gladly will demonstrate one of these
new Colorimeters—or, upon your re-
quest, the factory will send to you com-
plete descriptive information. Write

Dept. U8-4.

Spencer Lens Company

SPENCER

(o}

BUFFALO, NEW YORK
Scientific Instrument Division of

AMERICAN OPTICAL COMPANY

Sales Offices: NewY ork, Chicago, San Francisco, Washington, Boston, Los Angeles, Dallas,Columbus,St.Louis, Philadelphia, Atlanta

THE COLEECMING NE

[ Vor. XVI, No. 142

)

ITH you, as with us,
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Vol. XVI, No. 6

EFFECT OF SEA WATER ON THE RADIO-
SENSITIVITY OF ARBACIA SPERM
Drs. T. C. EVANS AND J. C. SLAUGHTER
Departments of Radiology and Zoology,
State University of Towa

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-

SATURDAY, AUGUST 2, 1941

Annual Subscription, $2.00
Single Copies, 30 Cents.

STRUCTURE OF THE RED CELL IN THE
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 150 years
after the discovery of the cell

jured more by the radiation
than were sperm irradiated in
concentrated suspensions. This
observation was checked by ir-

AU. DB. EF. Calendar

TUESDAY, August 5, 8:00 P. M.

by Swammerdam and later by
Leeuwenhoek) there has been
an almost continuous specula-
tion as to the reason why this

oe Be Seminar: Dr. Victor Schechter: | :
radiating sperm in different “Aging Phenomena, and Factors | Special shape should be as-
concentrations of sea water Influencing the Longevity of sumed by bodies floating free-

and later determining the fer- D
Yr.

Mactra Eggs.”
Frederick S. Philips:

ly in a liquid. Generally

“Com-

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

Dr. Matilda M. Brooks:

parison of Regional Respiratory
Rates of the Chick Embryo dur-
ing Early Stages of Develop-
ment.”

“Further
Interpretations of the Effects of
CO and CN on Oxidations in Liv-
ing Cells.” *

FRIDAY, August 8, 8:00 P. M.

Dr. P. B. Armstrong: “Function in

the Developing Gastro-Intestinal
Tract of Amblystoma punctatum
in Relation to Embryonic Deter-
mination and Differentiation.”

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

was added to the sperm their

into the structure of the color-

radiosensitivity increased. The
results of one of these experiments are shown in
Table I.

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
TABLE OF CONTENTS
Structure of the Red Cell in the Light of The Influence of Hormones on the Differen-
Shape Transformations, Dr. Eric Ponder.... 101 tioren of Melanophores in Birds, Dr. H. L.
amiltony Sele ee ees Se ei ree
Effect of Sea Water on the Radiosensitivity Conference at the University of Chicago.
of Arbacia Sperm, Dr. T. C. Evans and Ttemispof Interest icncsccesscescosscee eee
JC wslaughtenm... one te ee 101 Studies on the Life History of Siphodera
3 ve y vinaledwardsii, a Trematode Parasite of the
Some Aspects of Pigment Deposition in Toadfish, Drs. R. M. Cable and A. V.
Feather Germs of Chick Embryos, Dr. Ray FIM NMIMEN aeeseeeccceeesee cero ‘
Tig WVEMEESTESIOLY. coeeeepeeccren eee eee eee 107 Invertebrate Class Notes

— —— =s ee renner eee

‘aanjoid ey} JO Je4yxenb puey-qysi4
geddn ey} FO ToyUSD oY} UL axe selIOZELOGE] [VoISO[oIq ey, “pUnorseLoy ozerpeutUIE oy} UL ToqrIeyy 9]990T pus ‘paeK Aong oj “Quiog szedrune yyIM
HIOH SGOOM HO MUIA ANVITdalV

Aucust 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 10u 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
against each other, the two sets of forces maintain
the discoidal form. It is not difficult 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 p; and p» at each point, and computing the
pressure P which would have to be applied to
keep a homogeneous cell membrane, with tension
T, in hydrostatic equilibrium:

P = T(1/p1 + 1/pz).

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
exist is, of course, untenable, but the “outward
pressure over the equatorial regions’ is the same
thing as Gough’s “expansive force’. What really
happens 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

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under the Act of March 3, 1879, and was re-entered on July 23, 1938.
It is published weekly for ten weeks between July 1 and September 15 from Woods

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104

THE COLLECTING NET

[ VoL. XVI, No. 143

what a formulation of Norris’ theory might in-
volve.

The first problem, 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, with 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 be a product of
gelation, for the red cell has been shown to con-
tain proteins other than hemoglobin, e.g., the stro-
matin of Jorpes (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”
or “envelope” and the cell “membrane”, the for-
mer being at least thick enough to be visible
(0.05), while the thickness of the latter has been
put 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
Norris’ 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. Disc-sphere transformations.

As 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.

1. The spherical form between two 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
speculation 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.
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 any change in volume; they ultimately
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
probably itself about 14 A?. If the cells are sus-
pended 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

This sol keeps for weeks in the -

Aucust 2, 1941 |

the amount is needed at 37°C. as at 4°C., so the
temperature coefficient is positive, but not large.
(3) If the cells are fixed with fixatives such as
formol, 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 form produced by rose bengal.
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 brought 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 lysins.
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 process, 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,

THE COLLECTING NET

105

when the dise-sphere transformation occurs (Cur-
tis, 1935), nor of the electrical mobility (Furch-
gott and Ponder, 1940), which appears to be un-
changed, indeed, even after the cell is hemolysed
in several different ways (Abramson, 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 ).

3. The spherical form 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 apparently 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
about 60 per 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 membrane.

The study of the disc-sphere transformations
lead us to the conclusion that the changes are
probably due to the action of substances which act
at surfaces, and that the component which main-
tains the shape is probably 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
polarization optics have shown protein-lipoid com-

106

plexes in other membranes (e.g., the axon sheath,
Schmitt, 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
oriented 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 protein and of lipoid found
in the membrane by chemical means, there may
be 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 lipoid, 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
protein moiety would provide 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
different 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 phospholipoid 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 Dziemian,
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. at
pH 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

DHE COLLECIING, NET

[ Vor. XVI, No. 143

up about one-third of the protein content of the
membrane of the disc, is one such substance. I
therefore feel that it is possible 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 occupied 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-

Aucust 2, 1941 |
torted parts of the cell apparently “sphere up”
with difficulty, as if there were some restraint.
This is very clearly seen in the juvenile red cell,
the reticulocyte, in which bands of material, ap-
parently situated in the interior, can be 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 Waugh 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

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 réd 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 Biclogical 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% 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 morphogenetic 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

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 barbs
held together basally, and each barb bears 2 rows
of barbules. Each barb plus its barbules is desig-
nated collectively as a barb-vane. If these com-
ponent parts of the barb-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 barb-vane ridge projecting inward towards
the pulp and bounded peripherally by the feather
sheath. The barb lies at the apex, and one row
of barbules occupies 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 about the change from the
random distribution of pigment cells to a definite
distribution 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 barbs have not yet differentiated, and the cell
bodies of the pigment cells now lie at the apex of
each ridge. Their processes extend peripherally
and carry pigment to the barbule cells.

It is the nature of this relationship between the
pigment cell and the barbule cell which attracts
our attention. Previously the pigment cell has
been considered to be almost a micro-injection ap-
paratus capable of injecting pigment granules
into passive recipient barbule 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 become 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-

THE COLLECTING NET

[ Vor. XVI, No. 143

bule cells in a definite sequence. The melano-
phore 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
peripheral barbule cells are the first to elongate
and to form keratin, and only when these visible
differentiation processes begin can they receive
pigment. As this wave of differentiation spreads
progressively toward the pulp, the more axial
cells in turn become capable of receiving pigment.

(4) Pigment cell processes appear to be speci-
ically 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
plate 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 stimulate
the melanophores involved to undergo prolifera-
tion. If the apico-basal distribution of pigment
cells which are undergoing mitotic division is
plotted 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-
ly at 10 days and 18 hours of incubation, the germ
elongates rapidly, attaining its full growth by 18
days. 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
pigment deposition begins suddenly at that phase
of development when barbule cells begin to elon-
gate rapidly.

The down feather of Barred Rocks is solid
black in color, whereas the juvenile and adult

Aucust 2, 1941 ]

feathers of this breed exhibit alternate black and
white transverse bars. Willier has demonstrated
that 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

THE COLLECTING NET

109

cells from a regenerating feather germ, where they
would normally produce a barred pattern, are
transplanted into feather germs of White Leghorn
embryos, they deposit a solid black pattern 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 uvon 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. Howarp 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 types 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-
pionate, estradioi monobenzoate, testosterone pro-
pionate, 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-

(Continued on page 112)

110

THE COLLECTING NET

[ Vor. XVI, 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 founding 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,
round-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,
Henry Walters Professor of Zoology and Chairman
of the Department of Biology, Johns Hopkins Uni-
versity.

DATES OF LEAVING OF INVESTIGATORS

Baker, Gladys: «....s:...scccssscasccvssesssscesaeo eer eneenees July 21
lick, Sis. ‘De. scccccsscsossssssossssccstsgpeeaneesseeesseee eee July 23
Kreezer, G. ... . July 25
Millen sinuses .. June 30
Ronkiny Ri Re. ccsecscecissccvcctescsscecesessueeseeee tees July 25
Rothstein), cA... :..c..sc..ccesssscsoascssssesacceoe eee eeeeeeeteeeee July 14
Stowell, R. E. .. . July 27
Warner, E. N. .. .. July 25
Weaver, Margare go. densbucvcoueseceusucteceeneaeeeeeneeeee 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. Cincinnati. 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.
Gray, 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 following hours (Daylight Saving
Time) the current in the Hole turns to run
from Buzzards Bay to Vineyard Sound:

A.M.
1 dee ee EEO
. 12:41

Date P.M.

August
August
August
August
August
August
August
August
In each case the current changes approxi-

mately six hours later and runs from the
Sound to the Bay.

Aueust 2, 1941 |

THE COLLECTING NET

111

ITEMS OF

Dr. A. J. WATERMAN, who is instructing in the
invertebrate course, has been promoted from as-
sistant to associate professor of biology at Wil-
liams 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 Faculty 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. RatpH 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. JAMEes 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.

INTEREST

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.

GEE COOLER CMING NE

[ Vor. XVI, No. 143

STUDIES ON THE LIFE HISTORY OF SIPHODERA VINALEDWARDSII,
A TREMATODE PARASITE OF THE TOADFISH

Drs. R. M. Caste AND A. V. HUNNINEN

Purdue University and Oklahoma City University

Experimental studies on the life history of
Siphodera vinaledwardsu (Linton) have demon-
strated that this trematode is related to the Heter-
ophvidae as postulated by Manter, Price, and
Wilhelm on the basis of morphological and sero-
logical investigations. The definitive host in the
Woods Hole region is the toadfish, Opsanus tau,
practically all of which are infected. The small
marine snail, Bittiwim alternatum, serves as_ the
molluscan host in which the cercariae develop in
simple, elongate rediae. The cercaria is a pleuro-
lophocercous form of an unusual type since the
tail is inserted ventrally and coiled when at rest,
the fourteen penetration glands have two instead
of the usual four bundles of ducts in the region
of the oral sucker, and the excretory formula is

2[(2+2) + (2+2)] = 16 flame cells. The cer-
cariae penetrate and encyst in various species of
flounders, developing into infective metacercariae
in approximately 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
13-day metacercariae. Two of these have been
examined and found to contain large numbers of
very 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 them. 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
they become red or black melanophores, since both
kinds develop 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 example, fast-growing feathers
are mostly black in the Rhode Island Red breed,
so that physiological differences in feather germs
must favor the differentiation 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 i
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 by

stopping normal melanin synthesis at a red stage.
If this were true, all black melanophores should
pass 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 (1.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.

An 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
varying shades of brown, sandy, and red_ hair.
Dark brown and black hairs contain only black
melanophores. The similarity of these two kinds
of mammalian pigment 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-
phores 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.)

AvuGust 2, 1941 ]

THE COLLECTING NET

113

INVERTEBRATE CLASS NOTES

We invertebrates, fifty-five in all, representing
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 subject of three lectures
by 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 lab
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 be
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
of 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+lab, boiling un-
wanted lobster claws and playing bridge.

During lab hours (i.e., any time) we hear the
results of John Osmun’s newly (quite newly)
formed quartet with Bob Porter, Howy Miner,
and Sid Pond. The one person that will effec-
tively interrupt them is Gary Metcalf of the col-
lecting crew who never fails to praise the crew’s
soft ball 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 Batchelor

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

Conc. of sperm: “Dry” 135) lO
Time of insemi-
nation after
irradiation:
60 min. : 100 100 87
90 min.: 98 62 19

1:100 Diva 15 1:10 1:100
3 100 100 100 100
2 gy) 100 91 98

from the above 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 possibility 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 [ Vor. XVI, No. 143
Tas_e II.

Amount of radiation: None 1,160 r a \VA0) se 3,180 r 4,240 r 5,300 r

Percent. fert.: 100 69.5 45.0 35.4 41.5 33.8

Cone. Sperm 1:2000

Percent. fert. : 100 7.0 73 3.5 13.2 7.95

Amount of radiation: None 1,195 r 2,390 r 4,780 r 9,560 r 19,120 r

Conc. of Sperm 1:10

Percent. fert. : 100 100 100 100 93.0 93.0

Conc. sperm 1:1000

Percent. fert. : 100 73.0 63.8 36.6 TEN 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. : 100 97.2 98.3 99.3 92.1 56.9

Conc. of Sperm 1:1000

Percent. fert.: 100 94.7 94.4 ZA? 6.7 0

Amount of radiation : None 5,600 r 11,200r 22400r 44,800 r 89,600 r

Cone. Sperm 1:20

Percent. fert. : 100 86.0 86.0 68.5 48.8 40.7

Cone. Sperm 1:200

Perct. fert. : 100 12.6 1.5 0 0) 0

of the sperm at various concentrations was made
in the following manner. Sperm were collected
in as concentrated a form as possible, such sperm
being 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 sperm was diluted to the
same concentration before insemination in order
that the same amount 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 differences due to variation in
radiosensitivity from one lot of sperm to another.
Therefore the values to be compared are those

obtained 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 experiments 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 been 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.)

Aueust 2, 1941 | THE COLLECTING NET

in still projection

At the left is the 100-
watt Model MK Delin-
eascope for 2" x 2"
black and white or color ODAY’S high standards of color projection

slides. c i
owe much to the many optical and mechanical

In the center is the a ike ae
improvements originated by Spencer Lens Com-
fan-cooled 300-watt P ie) yee

Model MK-3, also for pany.
2" x 2" slides.

Voice rinks ist the Spencer offers the utmost in brilliance and clarity

750-watt Model GK of screen images and in operating conveniences.
Auditorium Delinea-
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344 x 4” slides. iti ;
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detailed information upon request.

Spencer Lens Company

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and Chase’s Principles of

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Lucas’ Elements of Human Physiology

Lynn's Organic Chemistry

McDougall’s Plant Ecology, new (3d) edition
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Whillis’ Elementary Anatomy and Physiology

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THE COLLECTING NET

[ Vor. XVI, No. 143

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Ten figures cover the stages in animal
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Ask for a copy of the May, 1941 Turtox
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Aucust 2, 1941 ].

MATE COLEEELING INET 117

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Position available in Preserved Material

Cooperation with
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State education, age, experience in preserv-
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quently write to us to ask if we will loan other preparations, salary desired, references,
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! Authors of biology texts and manuals fre-
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righted Turtox photographs and drawings

have been published in dozens of outstand- a ea i i i ah a
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[ Vor. XVI, No. 143

118 THE COLLECTING NET

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Main Street D Woods Hole

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The Cambridge Spot Galvanometer provides a complete
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Sensitivity in mm. on scale is from 19 to 170 per micro-
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| Cambridge makes many other types of galvanometers.

Cambridge Instruments are used to measure . . . minute direct and alternating potential,

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Aucust 2, 1941 ] 119

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120

THE COLEERELING Nba [ Vor. XVI, No. 143

ITH you, as with us,
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We will endeavor to give our
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All this is made possible because of the Bausch &
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To the pupil in the classroom, to the scientist
working with precision optical instruments and to
the wearers of Bausch & Lomb eyewear, the Bausch
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FOR NATIONAL DEFENSE, EDUCATION, RESEARCH, INDUSTRY AND EYESIGHT CORRECTION.

Vol. XVI, No. 7 SATURDAY, AUGUST 9, 1941 RETA S ERT

EFFECT OF AZIDE ON CYPRIDINA PROTEINS IN ACTION
LUCIFERIN Dr. Dorothy Wrincu
Dr. AurtN M. CHASE Professor at Amherst, Snuth and
Physiological Laboratory, Princeton University Mount Holyoke Colleges
Luciferin is the substrate in the bioluminescent One of the most interesting characteristics of

reaction. In the presence of an enzyme, lucifer- this golden age of biological research is the way
ase, and of oxygen in solution, light is produced. in which a new synthesis of ideas is emerging,

Reversible oxidation of luci- as the result of the immense
ferin can occur in the absence See ee srogress which has been
of luciferase but there is no 4. B. £. Calendar ee by specialist investi-
luminescence in this case. (E. gators working in a score of
N. Harvey, Ann. Rev. Bio- TUESDAY, August 12, 8:00 P. M. highly technical biological
chem., 1941.) Seminar: Dr. W. Trager: “Studies fields and the rise of the new

The luciferin and luciferase on Conditions Affecting the Sur- science of structure chemistry.
used in the present experi- uve) in vitro of a Malarial Para- | The original barriers between
ments were obtained from the Dr. Charles Hassett: “The Effect morphological and functional
crustacean, Cypridina hilgen- of Dyes on the Response to Light studies are giving way before
dorfu. In the thoroughly dried in Peranema.” font 0 the realization of the fact that
organisms the luminescent ma- oe 0. Habehons . Uunzerion of function can only be under-
terials remain stable practical- enn ect anos) | wectood im) termsyoimstructine mse
ly indefinitely. The luciferin Miss Virginia Dewey and Dr. G. that structure, far from being
was extracted and purified by W. Kidder: “The Possibility of | the concern only of certain
the method of Anderson (J. Thiamine Synthesis by Ciliates.” | specialists, is seen to be of the
Gen. Physiol., 1935) and the first importance in a_ great
luciferase was partially puri- FRIDAY, August 15, 8:00 P. M. variety of attacks on biological
fied by prolonged dialysis | Lecture: Dr. Franz Schrader: “The | problems. Biological systems
against distilled water. The see Movements of Chromo- indeed are now seen to be

reaction mixtures, of pH 5.4 superlatively fine examples of
and 6.6, were made up from functional architecture. Simul-
phosphate buffer, 0.2 M. and the sodium azide taneously structure chemistry, the science which
solutions were also made up in these same buffers. studies the geometry of intra- and inter-molecular
Luminescence was (Continued on page 131) atomic arrangements, has come of age. This

TABLE OF CONTENTS

Proteins in Action, Dr. Dorothy Wrinch........ 121 Dates of Leaving of Investigators «0... 128

Effect of Azide on Cypridina Luciferin, Dr. [tems oielnterestaeccsccscccctremccetteee eee ceseeeteserccnccd 129
PASI OM ASC: ce ceecratt seca desdcavcsatestssttvthesieacesvecsebeseee 121

Nominations for Trustees ........cccccccscccssseeeeeeereees 128° Invertebrate Class NOte@S) livtececsesccsceseccesonteseoeteee 130

Review of “Embryology of Insects and My-
Additional Investigators .....cc:cccccceeeseeeseeseees 128 MIapOU Sea eters Graiyaencccesste secretes ttceetesetess 13

= ee | AA

ALONG WATER STREET DURING THE 1938 HURRICANE

Above: The M.B.L. Club House at the height of the storm, with the Woods Hole Oceanographic
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)

Auecust 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

Tur CoLLEctINa 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 published weekly for ten weeks between July 1 and September 15 from Woods

marine biological laboratories.

| Hole, and is printed at The Darwin Press, New Bedford, Mass.

i Mass. Single copies, 30c by mail; subscription, $2.00.

It is devoted to the scientific work at

Its editorial offices are situated in Woods Hole,

124

THE (COLLECTING NET

[ Vor. 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 enzymes which operate through a
prosthetic group depend for their specificity on the
protein portion of the molecule. Further, several
vitamins have been 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.,
thyroglobulin. The sterol hormones are an im-
portant group for which evidence of protein com-
bination is lacking, but 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 genic 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 by 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 normal 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 substances.”
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
physical chemistry of proteins, studies of anti-
bodies and the crystallography of native proteins.
While 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

|

Aucust 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 suggested, 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 my 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
(if 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-
active native proteins, I propose to assume that
the proteins in cytoplasmic and membrane struc-
tures are in the latter form. It may 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
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 Bailentine 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 incidentally 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 by 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 delicacy, 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

DHE COLEECDING NET

[| Vot. 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 immunological facts. How otherwise,
indeed, can we interpret the subtle differences in
the affinities of antibodies formed by an antigen
formed from 1-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 Marrack 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 even 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-
ibility, 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. Any 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 suffh-
ciently well to form a stable combination, is neces-
sarily 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 Will 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 brought 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, Proc. Phys. Soc. London, 51, 592,
1939; Wrinch, J.4.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
different 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-

Aucust 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 different 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 same 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

[| Vor. 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 8, 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
auditorium of the Laboratory 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 Corporation — 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
Trustee of the Class of 1943 to replace W. J.
V. Osterhout—G. H. A. Clowes
Membership 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.

Bongiovennt, A. M. grad. biol. Villanova. Rock 3.

Bowser, E. Jr. asst. fel. biol. Pittsburgh. Rock 7.

Bowen, E. Bee biol. Gettysburg (Pa.).

Deyrup, Ingrith grad. zool. Columbia. OM 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, Marperet R. grad. biol. Brown. Br 312.

Memhard, R. (Riverside, Conn.). Br 334.

Nash, J. F. et 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
Phys. WF.

Reollason, H. D., Jr. grad. asst. biol. Williams. OM
Phys. Dr 7.

Sevag, M. G. asst. prof. biochem. Pennsylvania Med.

For

For

DATES OF LEAVING OF INVESTIGATORS

Costello, D. P. ... Aug.6 Mitchell, P. H..... Aug. 2
IOnbesieeeeseceee Aug.2 Mullins, C. P. ...... July 29
Hager, R. P. ......July 31 Runk, B. F. D..... July 26
Ele niy ul vapeerersescests Aug.2 Schaffel, M. ........ July 29
Hunninen, A. V.... Aug.9 Spratt, N. T., Jr. July 29
IGEN Abs LDS cosososntoa July 29. Warner, E. N..... July 25
REPRESENTATION BY i= AT
THE M. B.

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
Pennsylvania
New York ....

1940

Columbia ..
Yale
Hopkins ....
Harvard
Ohio State ..
@hicagoneeccsss erccrs
Rockefeller Institute
Michigan
Princeton ..
Brown
Cornell
Pittsburgh
Cincinnati
Lilly Laboratories ...
Milton Academy .....
Vassar
Villanova
Washington
California ..........
California Tech.
Iowa
Mt. Holyoke ...
Queens
Rochester :
GUiCs Ne) Yi:
Illinois
Miami
Minnesota ....
Missouri
Oberlin
Syracuse
Temple ......
Toronto .
Union

prowtuaceceserontsasanestsecensetee eee 30 34

23 21
18 22

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
August 10 .
August 11
August
August
August
August
August
August

A.M.
6334
7:18
8:01

P. M.
6:54
7:40
8:27
9:15
10:08
11:02
11:58
IZES

In each case the current changes approxi-
mately six hours later and runs from the

Sound to the Bay.

Aucust 9, 1941 |

ITEMS OF

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. Epcar J. BorLtt, who worked here last
summer, has been promoted from assistant to as-
sociate professor of biology at Yale University.

Dr. Franxk H. JoHNSON, instructor of biology
at Princeton University, has been promoted to as-
sistant professor.

Dr. Joun O. HutcueEns, 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 Marine 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 Club 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.

THE COLLECTING NET

129

INTEREST

PROFESSOR Otto LoEewt, who shared the Nobel
Prize in 1936, arrived in Woods Hole last week
with Mrs. Loewi. They will spend the balance
of the summer here.

Dr. JAMes E. Kinprep, 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. Davin Hann, 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. Mitprep W. S. SCHRAM, executive secre-
tary of the International Cancer Foundation, who
worked at the Marine Biological Laboratory last
year, 1s 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. Linvitte 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 staff. 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 held in the Old
Lecture Hall next Wednesday at 7:00. A pro-
posal to amend the by-laws to permit the mem-
bers of the immediate families of M.B.L. Corpor-
ation members to hold office will be considered.

130

THE COLLECTING NET

[ Vout. 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
abundant mimeographed directions one can select
material of particular interest. Our work on
Coelenterates included a study of the transparent
sea anemone Nematostella, 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.

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. While 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 Weber 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
bow! To make a long trip short, the suspecting
Hilton arrived at 8 P. M. and towed them home.

—Louise Gross and Bill Batchelor

BOOK REVIEW

JOHANNSEN, O. 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 pathological, 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-

Auecust 9, 1941 |

THE COLLECTING NET

131

ures 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 (J. Cell. Comp. 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, J. Cell. Comp. Physiol., 1936). An-
derson has also called attention to the fact that its
oxidation-reduction potential is similar to those
of certain naturally occurring polyhydroxy ben-
zene derivatives studied by Ball and Chen (J.
Biol. Chem., 1933). The absorption spectrum of
luciferin solutions (Chase and Giese, J. Cell.
Comp. Physiol., 1940) bears a marked qualitative
resemblance to those of certain naphtha- and an-
thraquinone derivatives measured by Morton and
Earlam (J. Chem. Soc., 1941). The visible ab-
sorption spectrum of luciferin solutions undergoes
changes during oxidation which may also indicate
a quinoid structure (Chase, J. Cell. Comp. Physi-
ol., 1940). Finally, Giese and Chase (J. Cell.
Comp. Physiol., 1940) found that cyanide com-
bines irreversibly with purified luciferin to give a
compound incapable of luminescence when lucifer-
ase 1s 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 (J. Amer. Chem.
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 HN» which
is effective, rather than the NaNs. 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
(Chem, Abstr., vol. 19, p. 3248), Franklin (J.
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 (J. 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 MNS, COMMACIMUNG, INNTA [ VoL. XVI, No. 144

To be ready for fall use—

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A SURVEY COURSE FOR COLLEGES
Edited by Gerald Wendt, Ph.D.
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CHEMISTRY. By Gerald Wendt. The book covers concisely atoms and molecules, elements and
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EARTH SCIENCES. By J. Harlen Bretz. Under the major headings of earth, water and air, this
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ASTRONOMY. By Clyde Fisher and Marian Lockwood. A short, informative survey of the Earth
as an astronomical body, of the solar system and the relations of its members to each other, and
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BIOLOGY. By Howard M. Parshley. The structural organization and chemical composition of
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134 THE COLLECTING NET [| VoL. XVI, No. 144

DALEY’S

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Aucust 9, 1941 }

THE COLLECTING NET

135

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136

Half a Glass Bead

OUR by hour her temperature has risen while

the doctor pits his skill and knowledge
against the merciless infection. But now the blood
count indicates that the infection has been checked.
The crisis is past.

Such drama is a 1941 commonplace. Through
the magic eye of the microscope, medical science
has learned more of the nature of disease and its
cure. Through “half a glass bead”—a tiny hemi-
sphere of optical precision—science looks beyond
superstition and ignorance, to see life processes
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With Bausch & Lomb’s pioneer application of
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century rarity—has become the twentieth century

[ Vor. XVI, No. 144

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Vol. XVI, No. 8

SATURDAY, AUGUST 16, 1941

Subscription, $2.00
Copies, 30 Cents.

Annual
Single

INTERPRETATIONS OF EFFECTS OF CO

AND CN ON OXIDATIONS IN CELLS
Dr. Matitpa M. Brooks

Research Associate, University of California

I. Effects on Rate of Oxygen Consumption.

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 Os 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.
(Continued on page 150)

It was found that

THE ORIGIN OF GAPS BETWEEN

SPECIES

Dr. Ernst Mayr

LV CW

A. B. E. Calendar

MONDAY, August 18, 8:00 P. M.
Lecture: Dr. R. W. Wilhelmi: ‘“Ser-

clogy Applied to Problems In-
volving Phylogenetic
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.”’

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.”

THURSDAY, August 21, 8:00 P.M. |
Lecture: Dr. H. Dam: “The Biolog-

Relation- |

from the question:
question: How does evolution proceed ?
the question on which the student of evolution

American Museum of Natural History,

p70)

Vork, N. Y.

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
field 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,
comparative 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

Is there evolution? to the
This is

TABLE OF CONTENTS
The Origin of Gaps Between Species, Dr. Proteins in Action (Cont.), Dr. Dorothy
PBSrana'S al Very tee eestesenstncteeeccaccsasssssascesuscacsicacssacenecer= 137 WINCH oa ees sess eeeecsees eee eeceneeeeceeeenneeneceseenccescencets 143
Interpretations of Effects of CO and CN on Biology at the a crSiLy, Olemlexals seeeemeeeete 146 |
Oxidations in Cells, Dr. M. M. Brooks........ 137. Additional Investigators ..........cceeeeseeeeeees 146 |
Aging Phenomena and Factors Influencing Dates of Leaving of Investigators.......0...... 146
Longevity in Mactra Egg Cells, Dr. Victor Items of Interest .
IS Chie Chik Igmeeeeete ne teeta tartan nceestesncecuanace snaarsvomacerssnee 143 Invertebrate Class Notes .........0...::ccccscscseeeesseeseee 148

Aucust 16, 1941 }

THE COLLECTING NET

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 prob-
lem of evolution, 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

139

the naturalist and ecologist, by the geneticist and
taxononust.

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-
brid 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-
ority 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

Tue CouuectiInc 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 published weekly for ten weeks between July 1 and September 15 from Woods

marine biological laboratories.

Hole, and is printed at The Darwin Press, New Bedford, Mass.

Mass. Single copies, 30e¢ by mail; subscription, $2.00.

It is devoted to the scientific work at

Its editorial offices are situated in Woods Hole,

140

Ti: (COLEECLING NEw

[ Voy. XVI, No. 145

species. The same is true, with few exceptions,
whenever we compare species that occur at the
same localities, 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 Rocky
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 Rocky Mountains. Here we
find 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 populations. 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 objective 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-
pletely separated species or incipient species. We
have learned from this discussion that species can
be delimited from two kinds of other species, from
sympatric species, where the gap is complete, and
from allopatric species, where the gap 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 species. 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 “obscura”
and D. “affinis.” The species of the flycatcher
genus Empidonax 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-
portance 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-
phological one. The sibling species around the
Malaria mosquito, Anopheles maculipennis, are
perhaps the most instructive illustration of this
principle. All the various races described by
earlier workers, such as atroparvus, messeae,
melanoon, sacharovi 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
quoted as evidence in support of the idea of sym-
patric 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
proceeds by geographical variation.

Geographical variation

Let us now examine the gap that exists between
allopatric, that is, geographically representative
species. 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
hybridization wherever such species ranges meet.
Morphologically, such forms are species, biologic-
ally they are not. These groups of allopatric spe-
cies are the final results of a process which we call
geographical variation. Let us now examine the
beginnings of this process.

Aucust 16, 1941 ]

THE COLLECTING NET

141

If a number of populations of any widespread
species are compared with each other, it is usually
found that they show certain genetic differences.
This has been proven abundantly by the work of
Sumner and Dice on Peromyscus, of Goldschmidt
on Lymantria dispar, of Dobzhansky, Timoféeff,
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 differences 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
subspecies are distributed continuously as in the
case of the mainland races of /yiolestes, 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 Myiolestes.

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 isolated 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 parent population, it is a species. But such a
purely morphological species definition is not ac-
ceptable to the modern biologist, because species
are, after all, not the creation of museum taxono-
mists, but products of nature.

We should, therefore, let nature make the deci-
sion what is to be considered a species, and not
rely entirely on the subjective judgment of the
taxonomust. 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
meet in nature and act like two different species,
that is they do not interbreed, we must consider
them as good species. A species definition based
on these biological criteria would have to be for-
mulated about as follows: ““A species consists of

a group of populations which replace each other
geographically or ecologically and of which the
neighboring ones intergrade or interbreed wher-
ever they are in contact, or which are potentially
capable of doing so in those cases where contact
is prevented by geographical or ccological 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 differ-
ences will eventually lead to a difference suffici-
ent to prevent interbreeding.

What the taxonomist is interested in is the
purely descriptive fact that the isolation of certain
populations 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
been 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
approach 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

THE COLLECTING NET

[ Vor. XVI, No. 145

vary - gaaraysisteelll 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, Bombus and Zosterops, are subspecies
or species. Geographical variation coupled with
isolation, completely blurs the borderline.

(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) Shght 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 exist 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.

Sympatric Speciation
I have treated speciation, up to this point, en-
tirely from the point of view of the ornithologist.
We 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 specializa-
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-
cificity 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 currents 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 different. 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 behavior, 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 unsolved 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 |

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
hierarchy of separate groups, of which at least the
species has an objective reality.

The ornithologists, and to some extent also the
students of mammals, butterflies and beetles, have
gone a long way toward understanding how the

THE COLLECTING NEY

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 are 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 New York

The unfertilized egg cells of the clam, Mactra
solidissima, 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-

PROTEINS

sium can easily be thought of as due to the dis-
placement of calcium in the egg cortex. It is also
known that 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 arabic; 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

ee 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-
cosity, pH, iso-electric points and salt binding
power are colligative properties.

Although many interesting parallelisms may be
drawn between growth, 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.

ego

(This article is based upon a seminar report pre-
sented at the Marine Biological Laboratory on
August 5.)

IN ACTION

Dr. DorotHy WRINCH
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

ANsls, (COMM RXCAMUNG

NET [ Vor. XVI, 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 EO: NH
groups, similar interlinks between acidic and ale
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 div alent ions of opposite sign.
It is well-known that hydrogen bridges have a
definite energy, say + to 8 keal. /mole. Contrast-
ing this with the energy in a carbon-carbon bond,
which we may take as (say) 60 kcal./mole, we
see that such bridges 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 bond. The nature of the
treatments by which such protein particles as
horse hemoglobin, the seed globulins, the hemo-
cyanins 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
veaker, namely that due to van der Waals attrac-
tions between hydrocarbon groups, such as exist
in hydrocarbon crystals. Such association will
depend upon a favorable opposition of what we
may call the van der Waals outlines of R-group
constellations on opposed 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 they are considered in terms of individual pairs
of R-groups and if our norm is the covalent bond,
but a sufficient amount of simultaneously opposed
groupings permits an association of considerable
stability. Next, in both cases the specificity of the
protein 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 specific 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
stable 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
exigency imposed by the polar groupings. Crystal
analyses have 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 Waals type of association. We can pic-
ture methyl groups so arranged that those from
one protein surface pack into pockets of methyl
groups from any other protein surface and wice
versa, by means of a kind of cushioning effect,
such that the surfaces can be pressed rather closer
together by means of the spreading of methyl
groups, 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 -CH»- 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. With
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

Aucust 16, 1941 |

case of cushioning associations of this kind there
will be no pores of definite size defined by rigid
atomic boundaries. The penetration of molecules
by 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 offered 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 slight preliminary discussion of per-
meability 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-
links. With this picture, we can visualize systems
which have wholly different specificities in virtue
of different patterns of interlinking, even if there
is little 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 (I 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 fundamentally
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

tHE (COLLECTING NET

145

be able to show, to the picture in the minds of
many biologists during the last eighty years. This
protein unit is large, but 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 1s 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 arrangement 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
by 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 erythrocyte systems 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 experimental 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, areal
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

HE OLERERING ENE

[ Vor. 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 38, 1879, and re-entered,
July 238, 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
members 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
University laboratories was opened to staff men
from hospitals, clinics and county health units.

The Clayton Foundation of Houston awarded
this 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 discoverer 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. McCullough of Detroit, Mich.,
and Leo Dick of Marshfield, Wisc., 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....... Henry, R.
Brooks, M. M Horn A@Bee
Bullock, T. H... ‘ Klotz, dine
Costello, D. O....... Aug.6 Mitchell, P. H.
Dumm, Mary E.....Aug.8 Plough, H. H..
Dytche, Maryon....Aug.7 Wolf, E. A...... F
HoT DES swwlayersseceecerece Aug.2 Woodward, A....... Aug.
Gilman, L. ............ Aug.11 Yntema, C. L....... Aug. 13

Dr. R. RuccGLes Gates is rewriting this sum-
mer at the laboratory his book on “Biological
Botany,” which was to have been published by an
English firm last fall. The typewritten manu-
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 December. 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 book included such topics as chlorophyll,
photosynthesis, stomatal action, rare elements in
metabolism, hormones, vitamins, viruses, X-ray
patterns in relation to cellular structure, chromo-
some structure, etc., embodying much recent work,

Dr. Joun Buck, 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
Johns Hopkins University, under the auspices of
the Brooks Fund, is also on the island studying
the development of frogs which 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
been converted into a laboratory and which has
been 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 AS P.M.

August 17 ; Asis}
August 18 lZe0 i 0y/
August 19 = le42e Sea
August 20 230) 245
August 21 SEO 3:30
August 22 4:01 4:18
August 23 4:45 5:03

Aucust 16, 1941 }

ITEMS OF

At the meeting of the Corporation of the Ma-
rine Biological Laboratory on Tuesday Drs. G.
H. A. Clowes, S. C. Brooks and Columbus Iselin
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 Cor-
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. Epmunp V. Cowpry, professor of cytolo-
gy at the School of Medicine of Washington Uni-
versity in St. Louis, has been appointed head of
the Department of Anatomy, succeeding Dr. Rob-
ein). Lerry.

Dr. R. W. GerarpD 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. Cart Hercet, who worked at the Marine
Biological Laboratory two years ago, has been
appointed instructor in physiology at Cornell
University Medical College.

Miss Dorotuy Dotr, 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 mixed 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.

THE, COLEECIING NET

147

INTEREST

Dr. Kennet S. Corr, associate professor 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. Grorce 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 McINNiIs, 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. Witper, 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. Ropert 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 Wednesday, 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, because 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 COLE GING

NET [ Vor. XVI, No. 145

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
species 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 lab 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 explosively, 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
Winifred whistled herself hoarse before the last
team reluctantly gave up 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
ferry the entire group. Supper of hamburgers

PROTEINS

INVERTEBRATE CLASS NOTES

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 Bill Batchelor

IN ACTION

(Continued from page 145)

cal problems, is of course a linear pattern 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.
When 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 about 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

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 & 10* units the size of insulin
per square micron, so that a protein complement
of the order of 107” 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 il

AN ete COLLECTING NET

in which lone Titkeavier filyaile are anes in (say)
four-way native protein units (topologically of the
type of the diamond structure) give water com-
plements of 70 to 99 per cent, corresponding to
fibrils 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 membranes 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.

149

The Een im eter Dees SS

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
Brtcke 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 Pfluger’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

hike COLEP GLRING NET

[ Vor. 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-
posed to have their seat in these fundamental ele-
ments. They make up all living matter, are the
bearers of heredity and the real builders of the
organism.”

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
experimental 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 protein structure in the last resort.

To my way of thinking, the idea in the mind
of 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
Svedberg, 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, [ 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
break 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 been 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
establish 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
by attempting to understand it and that others
should study its implications by experiment and
observation 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
problems.

(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)

different effects upon the rate of Oz consumption
were produced, depending upon the stage of de-
velopment. The results were as follows:

Methylene blue produced a rate of 130% in the
unfertilized stage of Arbacia as compared with the
control; 180% in the first cleavage stage; a slight
increase in the pluteus 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 Alsterias 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-

Aucust 16, 1941 |

WANS, COMIC WING, IN AL

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] 50

’

[Ox] 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 Os
consumption at different 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 different in
different stages of the same animal. If the redox
potentials are different, then the enzyme 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, 1f 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, Fett = FRet++, which must occur for
respiration, remains mainly in the Fet* 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-
ples may be applied. If methylene blue raises the

redox potential, for example, of a component of a
respiratory system so that its ratio of oxidants to
reductants is 50/50, the rate of respiration should
be increased. If however the system affected has
its equilibrium ratios changed by the addition of
methylene blue 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 Ov
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 (Jour. Biol. Chem. 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.

Il. 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 egg 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.
Methylene blue has doubled the life of both Ar-
bacia and Asterias; it produced faster develop-
ment, it increased the size of the plutei in Arbacia
from 280 », an average size in the controls, to
420 p» in length.

(This article is based upon a seminar report pre-
sented at the Marine Biological Laboratory on
August 5.)

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THE UTILIZATION OF AMMONIA BY
CHILOMONAS PARAMECIUM

Dr. JoHN O. HutTcHENS

The Johns Hopkins University

Chilomonas paramecium is a cryptomonad flag-
ellate about 30 micra long and 15 micra wide
which was found by Mast and Pace (Protoplas-

ma, Bd. 29, S. 326-359, 1933)
and subsequently by others to
grow in a variety of solutions
using single amino acids, urea
or aimmonia as a source of ni-
trogen, and lactate, acetate,
formate, or COy as a source
of carbon. Table I (Solution
A) shows the composition of
one of the solutions used by
Mast and Pace and used by
me in earlier work (Hutchens,
J. Cell. and Comp. Physiol.,
W217, 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.

third days.

40. BH. ©. Calendar

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
bryo.”

FRIDAY, August 30, 8:00 P.M.

Lecture: Dr. Rudolf Schoenheimer:
“The Dynamic State of the Body
Constituents.”

They can be subcultured indefinitely,
however, if transfers are made on the second or
(Continued on page 170)

crowded.

THE OFFICIAL
MARINE BIOLOGICAL LABORATORY

Em-

MEETINGS OF THE

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
came 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 new 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
indebtedness and that its fi-
na>ces are in good condition.
The number 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
We may congratulate ourselves that 1
spite of wars and rumors of wars, investigation

The Official Meetings of the Marine Biological

IR, Sb LP auilbifaS): concsoccecconcenacosacacbas ;
Studies on Conditions Affecting the Survival
in vitro of a Malarial Parasite (Plasmo-

TABLE OF CONTENTS

cal Laboratory, Dr. Charles Packard............ 157 Peranema trichophorum, Dr. C. Hassett.... 163
The Utilization of Ammonia by Chilomonas Program of the Summer Meetings of the Gen-
paramecium, Dr. John O. Hutchens .............. 157 etics Society of America at Cold Spring
Comparison of the Respiratory Rates of Dif- Harbors ongalsland Nem Yen 164
ferent Regions of the Chick Blastoderm Summer Activities of University of Chicago
During Early Stages of Development, Dr. BIO] O SISES. oe <8 ss ssee Reece eee ee

Items of Interest

The Effect of Dyes on Response to Light in

Invertebrate Class Notes cesses. 168
dium lophurae), Dr. W. Trager ............000 162 Some Recent Books in the Biological Sciences 168

ge

WIOH SGOOM AO SUIMOLVUOAVT IVOIDO'IOIN ANIIVW AHL

Aveust 23, 1941 |

WiKNE, COMMLIRCIMUNE! INIT AL 159

and instruction have gone on almost unhampered.

The completion of the new wing of the Library
marks another stage in the development of this
important part of our institution. In earlier days
it was housed on the Protozoology Laboratory :
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. [f
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, rubber 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 words:

“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 own opin-
ion that it is improbable that any such demand
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 Chemistry,

Vale 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 Nev was entered as second-class matter July 11, 1935, at the Post Office at Woods Hole, Mass.,

under the Act of March 8, 1879, and was re-entered on July 23,
It is published weekly for ten weeks between July 1 and September 15 from Woods

marine biological laboratories.

Hole, and is printed at The Darwin Press, New Bedford, Mass.

Mass. Single copies, 30c by mail; subscription, $2.00.

1938. It is devoted to the scientifie work at

Its editorial offices are situated in Woods Hole,

160

DHE COLEECIING 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 between 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 respiratory 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 (White 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 Linderstr¢m-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. pl. Oy 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-
sumption 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 publish 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 Qo,’ of the
embryonic pieces could be calculated as_ the
m. wl. Oy consumed/hour/y 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. Oy 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 Qo,’ 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 Qo,’ values. The other
regions have somewhat higher rates, but from the
values of the standard deviations it does not ap-
pear 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 Qo,’ 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 Qo,’
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

,

Aucust 23, 1941 |

THE COLLECTING NET

161

TABLE I.

The Qo,’ (m. » 1. Oz consumed/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- Node and An- Middle Posterior Right Left
Process terior Streak Streak Streak Lateral Lateral
77 75 74 85 89 92
ze OS) SE 7/5 ==) 10/8 se 1165 + 198 + 199

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 11 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. Os 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-
bated blastoderm is of the same magnitude as the
value for the whole blastoderm 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. pl. in the
pellucida area of the unincubated blastoderm to
about 313 m. pl. 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 i
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
Janus green reduction under the induced anaero-
bic conditions of their experiments is the thickest

162

THE COLLECTING NED

[ VoL. XVI, No. 146

region of the head-process area pellucida. Such
a region ought to show the presence of the re-
duced form of the dye before 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 descriptive 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 Qo,, 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 TRAGER
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 sexual cycle of development. There
are 4+ species of malaria parasites of human beings,
no one of which has ever 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, Plasmodium lophurae, has been used for
the work to be 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
presented 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 im witro. 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-
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 promote either
some development or rapid death. Hence, all the
experiments were performed at temperatures of

Aucust 23, 1941 }

iit COLEECIING NET

40-42°C., about the body temperature of the
chicken. Now, how can we judge the survival of
malaria parasites? They are small and typically
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
+. 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.
lophurae 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

163

in a tube 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 COy, — 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 giucose 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 +-+ 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
parasites were alive on the 3rd day, at least 20%
on the 4th day, about 1% on the 5th day and
about 0.05% on the 6th day, the last day of the
tests. In such preparations there was a small in-
crease in parasite number on the first day of life
in vitro.

(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 INPERANEMA TRICHOPHORUM
Dr. CHarLes C. HAssETT
Department of Zoology, The Johns Hopkins University

Peranema trichophorum is a colorless flagellate
without an eyespot. It moves by swimming or by
crawling; when it crawls it moves slowly and is
tasily kept under observation. It responds to a
sudden increase in luminous intensity by ceasing

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

[ Vor. 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 Peranema 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-time to approxi-
mately 1 sec. when the concentration of dye was
2 < 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 * 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 & 10° M; more dilute solu-

tions produced shorter reaction-times down to
12 sec. at 1 X 10° M. This dye is much more
toxic 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 Peranema 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
Peranema, 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 Peranema
vary from auramine O, which absorbs light main-
ly in the region 3900-4300 A to methylene blue,
which absorbs 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 Peranema; (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. DopzHANSKY, Columbia University.
Vice-President, E. W. Linpstrom, Iowa State College.
Secretary and Treasurer, B. P. KAUFMAN, Carnegie In-

stitute of Washington.

Wednesday, August 27, 8:30 P. M., Blackford Hall,
Evening Lecture

A. H. Srurrevant, California Institute of Technology #
Comparative Genetics of the Species of Drosophila.

Aueust 23, 1941 |

THE COLLECTING NET

Thursday, August 28, 9:30 to 12:45, Davenport
Laboratory, Demonstrations

(1) Atwoop, SANForD, S., U. S. Regional Pasture
Research Laboratory, State College, Pa.: The multiple
oppositional alleles causing cross-incompatibility in Tri-
folium repens.

(2) BrsHop, D. W., University of Pennsylvania,
Philadelphia, Pa.: Cytological demonstrations of chro-
mosome breaks soon after X-radiation.

(8) BreHMe, KATHERINE S., Carnegie Institution of
Washington, Cold Spring Harbor, N. Y.: A survey of
the Malpighian tube color of the eye color mutants of
Drosophila melanogaster.

(4) BusHNELL, RatpH J., The University of Con-
necticut, Storrs, Conn.: Incompatible matings in inbred
families of the bean weevil.

(5) Drmerec, M., HOLLAENDER, 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) E1est1, O. J., University of Oklahoma, Norman,
Okla.: A comparative study of the effects of sulfanila-
mide and colchicine upon mitosis of the generative cell
in the pollen tube of Tradescantia occidentalis (Brit-
ton) Smyth.

(7) FANKHAUSER, GERHARD, and
Princeton University, Princeton, N. J.:
of spontaneous aberrations of chromosome
among larvae of the newt, Triturus viridescens.

(8) Gorpon, Myron, New York Aquarium, New
York, N. Y.: Dominant and recessive responses of the
Sd factor in natural and domesticated fish populations.

(9) Hrnton, O. TAyLor, and Arwoop, 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) Kamenorr, Raupu J., City College, New York,
N. Y.: A cytological study of the embryonic livers (16-
18 days) of normal and flexed-tailed (anemic) mice. ~

(11) Kimpatt, R. F., Johns Hopkins University,
Baltimore, Md.: A gene affecting the manner of swim-
ming in the ciliate protozoan, Huplotes patella.

(12) Laanes, T., and MacDoweEL., E. C., Carnegie
Institution of Washington, Cold Spring Harbor, N. Y.:
Screw-tail, a new mouse mutation.

(13) Power, MAxweLu E., Yale Universitty, New
Haven, Conn.: Neurological effects of mutants reducing
facet number in the eyes of Drosophila melanogaster.

(14) RippuE, 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) Roserrson, 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., MeGill University, Montreal,
Canada: Spiralization in microspore chromosomes of
Trillium. e

(18) Spencer, W. P., College of Wooster, Wooster,
Ohio: Inherited variations in wild populations of Clay-
tonia virginica.

(19) Wrutson, G. B., and BoorHroyp, E. R., McGill
University, Montreal, Canada: Differential reactivity in
the chromosomes of Trilliwm species.

(20) Wruson, G. B., and Sparrow, A. H., MeGill
University, Montreal, Canada: Partial fusion of un-
treated root tip chromosomes of Trillium erectum L.

CrotrtTa, Riva,
The frequency
number

Afternoon Session, beginning at 2 o’clock
INSPECTION OF EXHIBITS prepared by resident investiga-
tors of the Department of Genetics, Carnegie Institu-

165

tion of Washington and of the Biological Laboratory
of the Long Island Biological Association.

Late Afternoon and Evening
Swim, Picnic SUPPER, AND DANCE.

Friday, August 29, at 9:30 A. M., Auditorium of
Blackford Hall

(1) BerrnstTeIn, MAariANNE E., Carnegie Institution
of Washington, Genetics Record Office, Cold Spring
Harbor, N. Y.: The incidence and Mendelian transmis-
sion of mid-digital hair in man.

(2) LINDEGREN, Cart 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 genotype.

(4) Neset, B. R., Wiuson, G. B., and MarrNneui, L.,
New 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 Nese, 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) Caspari, Ernst, Lafayette College, Easton, Pa.:
Genetic and environmental factors influencing testis color
in Ephest.a kiihniella.

(7) Muturr, 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
Droscphi'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) Neetu, J. V., Dartmouth College, Hanover, N.
H.: A case of high mutation frequency in Drosophila
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
investigators registered at the Marine Biological
Laboratory each year since its foundation.

166

Dies COLEECIING NED

[ Vou. 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 238, 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
marketed. Dr.

Oldham will bring back pituitary

glands for her work with Dr. Eugene Geiling,
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. Stunkard, 6-0, 7-5

In the finals of mixed doubles, Lancefield and Te
Winkel defeated Lumb and Humm, 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 defeate:
G. Gorokhoff, 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. Muss Galina Gorokhoff 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!

Rejoice in the Lord alway
Ave Verum Corpus William Byrd
Gospodi Pomiluy (Lord, Have Mercy) Lyovsky
When His loud voice (Jeptha) Georg F. Handel

Brightly Dawns Our Wedding Day (The Mikado)
Gilbert and Sullivan
Orlando di Lasso
German Folk-Song

Alexander Kastalsky
Henry Purcell

Good-day, dear heart
The Beetle’s Wedding
A Legend P. Tschaikowsky
Round the Good Father’s Door A. Arkhangelsky
Moon Magic Three Russian Folk Songs

M.B.L. CLUB

The ping pong tournament of the M.B.L. Club
is well under way with most of the first and some
second rounds played off. 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 vane yard Sound:

Date P. M.
August 24 .. 5:30) ora0
August (25) 5.) OclommmOromg
August 26 ....... HAO © 4 e27/
UIC ISEN 2/7 7AD S29
INDE AS crest sil
August ZO 9 So ORIS
August 30 LOESS) SES

Aucust 23, 1941 |

ITEMS OF

Dr. Dororuy M. Wrinch was married on
Wednesday at 12:30 P. M. in the Laboratory to
Dr. Otto Glaser by Rev. Ralph H. Long of Fal-
mouth. They are spending their honeymoon on
Nantucket Island, and will return later to their
house in Quisset. Dr. Wrinch will continue her
scientific work as before; 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.

Katherine Brehme was her only attendant.
The marriage is the first 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. Menkin, Isabel M. Morgan and K. G.
Stern.

Mr. Davin 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. Gorrin, 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 number
probably was about four times as great.

The Anton Dohrn 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.

HEHE COLEECDRING NET

167

INTEREST

Proressor A. H. SrurtevAnr will give the
evening lecture at the summer meetings of the
Genetics Society of America at Cold Spring Har-
bor on August 27. He will speak on ‘“Compara-
tive Genetics of the Species of Drosophila.”

Mr. Rosert 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. JouHnson, assistant director of the
Bartol Research Foundation, has been working at
the Oceanographic Institution for brief periods
during the summer.

Dr. J. H. McGrecor of Columbia University,
left Woods Hele on Thursday after a two-day
visit.

Dr. Hartan 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.
whom were at the Laboratory in
Woods Hole briefly last week-end.

both of
visited

M. W. BoswortH,
1940,

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. Rogert CHAMBERS will leave Monday for
New York to attend a conference on tissue cul-
ture.

Mr. Cart 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. WooprvurFF 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. Epwarp 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 members
of the Laboratory and their families with a garden
party at which a Craig ae of a Russian fairy
tale was presented. Robert Chambers acted
as narrator ace ane play.

168

DHE COLLECDRING

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 trouble keeping us interested in his
discussions. Of particular interest in the lab
work were the dissections of busycon 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 teams,
but every one was glad to abandon 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 Nereis 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 watermelon-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 w ith 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 Batchelor

SOME RECENT BOOKS IN THE BIOLOGICAL SCIENCES

Ackerman, E. A
$4.00. Chicago.

Adams, N. E., and E. M. Bandow. A Study Guide
for Applied Biology. $1.75. Burgess.

New England’s Fishing Industry.

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.

Beaver, W. C. Laboratory Outlines of General Biol-
ogy (2nd ed.). $2.00. Mosby.

Bell, D 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. III, $3.50.
Jaques Cattell Press.

Blum, H. F. Photodynamic Action and Diseases
Caused by Light. $6.00. Reinhold.

LBrockleshy, 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
$3.50. Mosby.

Carlson, A. J., and V. Johnson.
the Body. $4.00. Chicago.

Health.
The Machinery of

Carroll, P. L., and W. P. Horner.
$1.25. 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. $3.00. Blakiston.

Cenn, 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.

Cress, 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
(vev. 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.
Enzymes. $3.25. Burgess.

Evans, H. M. Brain and Body of Fish. $3.50. Blak-
iston.

Fasten, N. Introduction to General Zoology. $3.75.
Ginn.

Felt, E. P. Plant Galls and Gall Makers. $4.00.
Comstock.

Federal 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.

Atlas of the Frog.

Respiratory

|

Aucust 23, 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.

Gause, 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 (8rd 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. Robbins.
Botany (8rd ed.). $2.75. Wiley.

Hoskins, R. G. Endocrinology; the
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. $4.50. 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.

Ley, W. The Lungfish and the Unicorn: an Excur-
sion into Romantic Zoology. $2.75. Modern Age
Books.

Longley, M. H. Laboratory Manual in Bacteriology.
$1.50. F. A. Davis.

Elements of

Glands and

Luck, J. M., ed. Annual Review of Physiology, Vol.
III, 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.).
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. $1.25. 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.
Biology. $1.00. Harcourt.
Smith, K. The Virus, Life’s Enemy. $2.00. Cam-

bridge (Macmillan).

Snell, G. D. Biology of the
$7.00. 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.

$3.00. Cassino

Guide to Modern

Laboratory Mouse.

170

Wald, GCOLMIACMONG INI

[| Vor. XVI, No. 146

Thorsteinson, E. D. New or Noteworthy Amphi-
pods from the North Pacific Coast. Washington.

Transeau, E. N., H. C. Sampson and L. H. Tiffany.
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.
Waddington, C. H. Organisers and Genes. $3.00.

Cambridge (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
Guide. $2.90. Burgess.
Wolcott, R. H. Animal Biology (2nd
McGraw-Hill.

Woodruff, L. L., ed. Development of the Sciences,
Second Series. $3.00. Yale.

Woodruff, L. L. Foundations of Biology (6th ed.
rev.). $3.75. Macmillan.

Zoology Laboratory
ed.). $3.50.

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-
plasma, 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
solution with the acetate omitted but with a gas
phase containing a partial pressure of COz 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 situation still obtains with
respect to the inorganic solution, but as shown by
Hutchens (J. Cell. and Comp. Physiol., V. 16, pp.
265-267, 1940) (and unpublished results) by use
of a solution to which thiamin and iron are added
populations up to 1 ae cells/ec. can be ob-
tained (Table I, Solution B In addition to the
B, and iron it will be ne 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 of Solutions Used for Growing
Chilomonas

Substances mg. %

Solution A Solution B
CH3COONa:3H20 249 249
NH,4Cl 46 46
(NHy)2SO4 10 10
K»HPO, 20 150
MegCls 1 1
CaCls 1 1
FeCls -- 0.17
Thiamin hydrochloride —— 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/ce. 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 obtaining of
additional energy by oxidation of ammonia.

The experimental methods used are as follows:
The organisms were grown in sterile, pure, mass
cultures in 125 cc. Erlenmeyer flasks containing
50 ec. of the solution shown in Table I (Solution
B). Inocula of 1-2 thousand organisms/cc. were
used, 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 different 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. Wet 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

Aucust 23, 1941 | THE

COLLECTING NET

1/1

TABLE II.

Wet and Dry Weights of Chilomonads During
the Logarithmic Phase of Growth
(48 Hour Cultures)

Wet Weights
mg./10° cells

Dry Weights
mg./10° cells

2.64 0.738
2.42 0.559
2.42 0.563
2.60 0.692
BOM 0.530
252 0.595
2.48 0.575
2.49 0.563
2.87 0.767
ellil 0.545
Zeal 0.567
2.48 0.567

average = 2.51

average = 0.605

Dry weight = 24% of wet weight.

the ammonia from an alkalized suspension of cells
and subjecting the residue to Kjeldahl digestion,
or differentially by measuring the ammonia pres-
ent before and after Kjeldahl digestion, both
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 HeSOg.

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 & 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 by 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 linge two Gomponents 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.
25y/cc. had disappeared. Subjection of the total
solution, 1.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 + & 10° organisms/cc. or
1 mg. of wet cells/ec. As seen from the previous
table, these would be expected to be 2.5 percent
N, i.e. to contain 25y 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 1n-
significantly small. The conclusion therefore is
that i in the solution used Chilomonas does not oxi-

TABLE III.

Nitrogen Content of Chilomonads During the
Logarithnuc Phase of Growth

mg. protein/10°

Hae NOT EET cells (N X 6.25)

0.056 0.35
0.071 0.44
0.060 0.38
0.066 0.41
0.063 0.39
0.066 0.41
0.063 0.39
0.063 0.39
average = 0.064 average = 0.40

Nitrogen (% of wet weight) = 2.5%
Protein % of wet weight) = 16%

172 THE COLLECTING NET [ Vor. XVI, No. 146
TABLE IV.
Changes in Nitrogen Content of Cultures of Chilomonas
Original Final Final Organic % of Original Final Original Final %
Expt. NH:;-N NH:;-N Total N Total N NO:-N NO.-N NO:-N NO;:-N _ recoy-
N Fixed ery
y/ ce. 7/ ee. 7/ce. y/ce. 7/ce. 7/ce. y/ee. 7/ce.
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 -— oa 100
113 146 33 226 0.01 0.01 — are
3 146 112 146 34 2353 0.01 0.01 — — 100
112 148 36 24.5 0.01 0.01 — — 101
4 150 ie) 149 30 20.0 no analyses 99
5 147 15 146 31 Zl 98
114 147 33 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 — — go)
dize significant amounts of ammonia, but all that gen. You will see that this amounts to 25-30 per-

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 be 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 Amide N N
mg. mg.
1 35 al BS)
2 Sil 8 26
3 18 5 28
+ 79 20 25
Ue 18 25
5 40 12 30
6 51 12 24
7 87 23 32
average = 28.2

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 been previously reported by numer- —
ous 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 ammonia in the
culture ‘solution.

2. It has been shown that in the solution used —

no significant amounts of ammonia are oxidized
to furnish energy for growth.

3. Finally it has been shown that of the nitro-
gen in the proteins a relatively high proportion 1s
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 on
August 12.)

4
.

Auecust 23, 1941 | THE COLLECTING NET 173

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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 implications following from
the flexible protein framework picture of cytoplas-
mic and membrane structures. I refer to (1) the
synthesis of proteins and (2) the function of me-
tallic ions in physiological processes.

(1) Ihave 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 be capable also of ac-
counting for the anomalous properties of cyto-
plasm; and that fabrics of such units may be the
basic structural units in biologically active mem-
branes (Cottectinc Nev, 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. MEYERHOF
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 physiology. 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 importance 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 labile 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.
OMMWievier Mote recs Lo scisecssdecessrceccssatsachcessasaveens 177

Further Implications of Flexible Protein
Frameworks, Dr. D. Wrinch ............0cc0cc0e 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 .............ccccccseccescoeeesseeee 184
Papers and Demonstrations Presented at the
General Scientific Meeting...................:ss0cesssee0 185
Books REWIGWS) erisyccccxscssocssoncenseencecccteceestoseneee 186
Academic Rank of M.B.L. Investigators 186
Seminarsvat Mountain Wakely vcscccccaserescescereeets 186
tbemsvot Interestsciisssscccececsesnccctecs costetseess sees eeescss 187
Memorials Read at Corporation Meeting of |
the Marine Biological Laboratory ................ 188

178

AEN, COMI CIMING: INI

OH OH
HN — P= HN — P=0O
| \ | N
| OH OH
5 = Niel 5 = INJal
7 ¢ (Clala 1
ee (CH2)s
COOH iia
COOH
Creatine phosphoric Arginine phosphoric
acid acid

FIGURE 1.

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 without
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, was 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. While 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-
sentatives 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 only 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 substituting 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 chemical 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

[ Vou. XVI, No. 1479

Aucust 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 Balanoglossus, an earlier class of chor-
dates than the more recent acrania amphioxus,
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 both 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 sub-
stance, which proved to be a condensation product
of arginine and propionic acid, called octopine.
This substance 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-

Creatine
phosphate

Arginine

Phylum and class phosphate

Most invertebrate phyla + ==
Echinodermata
Crinoidea +
Asteroidea +
Holothuroidea +
Echinoidea +
Ophiuroidea —

Protochordata
Tunicata -|-
Enteropneusta se
Cephalochorda —

+++

Vertebrata, 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 1odoacetic 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

Tur 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 published weekly for ten weeks between July 1 and September 15 from Woods

marine biological laboratories.

Hole, and is printed at The Darwin Press, New Bedford, Mass.

Mass. Single copies, 30c by mail; subscription, $2.00.

It is devoted to the scientific work at

Its editorial offices are situated in Woods Hole,

180 THE COLLECTING NET [ VoL. XVI, No. 147
ie
lel (CIN) OH OH
\
(lal OH
Ze |
ren aires CH—CH—CH—CH,.—O—P
| |
O
I. Adenylic acid = Adenosinmonophosphoric acid
N=C:-NHe
I
HC €—N OH OH
| \ Pl
CH | OH OH

Ye |
N—C—N—CH—€H—€H

| |
CH—€H;—0—P—O— Por

| |
© © oO
II. Adenosindiphosphoric acid
aetee
|
HC C—N OH OH
[ee eS | |
CH | lal ~@lel Ola!
ie lit leek. | | |
N—C—N—CH—CH—CH—CH—CH:—O— P—O— P—O— P—OH.
| | | |
o On” "Ox uae
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, which 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.

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 group 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

One of ©
these is explained by equations C and D. The last ©

|

Aucust 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 substances.

I may add some words about the distribution
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
much as in the same quantity of muscle; part of
this phosphagen breaks 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 creatinephosphate + adenylic acid =
2 creatine + adenosintriphosphate

B) adenosintriphosphate — adenosindiphosphate
+ H3PO 4 — adenosinmonophosphate

(adenylic ac.) + 2 H3PO,

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

FE) 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 undoubtedly 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 Ay. 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. NACHMANSOHN
Laboratory of Physiology, Yale University School of Medicine

Electrical changes during nerve activity occur
within a few milliseconds or even within a frac-
tion of a millisecond. 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 ACh
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 ACh metabolism 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 electricus (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
(92%) 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-
phorus 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 marmorata, 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

Aueust 30, 1941 |

THE COLLECTING NET

183

only a quantitative one. Bioelectric potentials are
surface phenomena. The localization 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., [= current and R = resistance) two as-
sumptions can be made about the way in which
ACh may act:

1) ACh can produce the E.M.F. directly by
action on the surface. This possibility 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 differences are responsible for
the potential differences.

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 for Medical Research, New York, N. Y.

The chemical nature of mitochondria, and the
function which they assume in the economy of the
cell, are problems 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 mp 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 be 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 Harbor, 1941; R. R. Bensley and N. L.
Hoerr, Anat. 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 3, 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

[ Vor. XVI, No. 147

by proper cytological techniques (R. Noél, 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 mp 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 pig 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 about 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 Noél (Arch. Anat. Micros., 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. Reviews, 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)

Aucust 30, 1941 ]

DHE COLLECTING NET

185

PAPERS AND DEMONSTRATIONS PRESENTED AT THE GENERAL SCIENTIFIC
MEETING, 1941

Tuesday, August 26, Morning Session, 9:00 A. M.

CASWELL GRAVE:
of Ascidian larvae.

CASWELL GRAVE: The ‘‘Eye Spot’’ and light re-
sponses of the larva of Cynthia partita.

LuoyD BIRMINGHAM: Regeneration in the
zooids of Amaroucium constellatum.

Ivor CORNMAN: Characteristics of the acceleration
of Arbacia egg cleavage in hypotonic seawater.

ErHEL Browne Harvey: Material inheritance in
Echinoderm hybrids.

E. 8. GUZMAN BARRON AND J. M. GOLDINGER: Inter-
mediary carbohydrate metabolism of sperm and eggs of
Arbacia before and after fertilization.

J. D. CRAwForD, D. Brenepict, A. B. DuBors 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. Minne: 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. Faruua, J. C. SLAUGHTER AND E. P.
LirtLe: Influence of the medium on the radiosensitiv-
ity of Arbacia sperm.

C. Lapp Prosser AND G. L. ZIMMERMAN: Compara-
tive pharmacology of myogenic and neurogenic hearts.

Lois E. TEWINKEL: Structures concerned with yolk
absorption in the dogfish, Squalus acanthias.

W. H. F. Appison: 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.

Further studies of metamorphosis

early

M. H. Jacops anp DorotHy R. StBwaRT: Catalysis
of ionic exchanges by bicarbonates.
DorotHy R. Stewart AND M. H. Jacogs: The role

of carbonic anhydrase in the catalysis of ionic exchanges
by bicarbonates.

Martin G. Nersky anp M. H. Jacoss: Some effects
of desoxycortico-sterone and related compounds on the
mammalian red cell.

HERBERT SHAPIRO AND HuGH Dayson:
of the Arbacia egg to potassium.

A. K. Parpart: Lipo-protein complexes in Arbacia
eggs.

S. E. Hitt: Relation between the action potential
and protoplasmic streaming in Chara and Nitella.

AuRIN M. CHASE: Observations on luminescence in
Mnemiopsis.

Permeability

Papers Read by Title

Frep W. Autsup: Photodynamic studies on Arbacia
eggs.

Ivor CorNMAN: Disruption of mitosis in Colchicum
by means of colchicine.

T. C. Evans: The effeet of roentgen radiation on
the jelly of the Nereis zygote.

R. RuGGLes Gares: Tests of nucleoli and cytoplas-
mic granules in marine eggs.

Russert P. Hacer: Sex-linkage of stubby (sb) in
Habrobracon.

E. R. Hayes: The Elasmobranch interrenal; a pre-
liminary note. The interrenal body of Alopias vulpinus
(Bonnaterre).

Dwicut L. HopKINs:
rucosa.

Grorce W. Hunter, III, anp EDWARD WASSERMAN:

The cytology of Amoeba ver-

Observations on the mealanophore control of the cunner
Tautogolabrus adspersus (Walbaum).

FLoreNce Moog: The influence of temperature on
reconstitution in T'ubularia.

CLintoN M. OssorN: Factors influencing the pigmen-
tation of regenerating scales on the ventral surface of
the summer flounder.

G. H. Parker: MHypersensitization of catfish melano-
phores to adrenaline by denervation.

LronarpD P. SAyLes: Implants consisting of young
buds, formed in anterior regeneration in Clymenella,
plus the nerve cord of the adjacent old part.

A. A. SCHAEFFER: Chaos nobilis Penard in perma-
nent culture.

VictoR SCHECHTER:
cells.

Sipney F. Venick: The effect of centrifugation upon
the oxygen consumption of Arbacia eggs.

ALLYN WATERMAN: LEctodermization of the larva of
Arbacia.

RaLPpH WICHTERMAN:
Paramecium bursaria,

Fioyp J. Wiercrnski: An experimental study of in-
tracellular pH in the Arbacia egg.

E. ALrrep Wo.ur, MaAryon DytcHE AND MILTON
ScHAFFEL: Heat produced by respiring whole blood of
Tautoga onitis and Mustelus canis.

C. L. Ynrema: Effect of differences between stages
of donor and host upon induction of auditory vesicle
from foreign ectoderm in the salamander embryo.

Further studies in Mactra egg

Studies on Zoochlorella-free

Wednesday, August 27, 2:00-4:00 P. M.
Demonstrations

L. F. Boss ann M. H. JAcops: 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. N. Harvey anp F. J. M. SicHeL: Apparatus for
intense flashes of ultra violet light, and the killing of
small organisms.

E. N. Harvey anp F. J. M. SicHEL: Apparatus for
high speed photography with the microscope.

Kurt G. STERN: An air-driven high-speed centrifuge
for optical observations.

Ivor CorNMAN: Disruption of mitosis in Colchicwm
by colchicine.

SEARS CROWELL: Nematostella, a simple anemone,
suitable for laboratory work in general zoology.

SEARS CROWELL: The nematosomes of Nematostella.

JoHN KrosiAN: Apparatus of simple construction
for use in microchemical work.

JouHN Krosian: A spot test method for the quantita-
tive determination of magnesium in tenths of a micro-
gram.

ELEANOR H. Suirer: A mutant Drosophila melano-
gaster with extra sex combs.

Lois E. TEWINKEL: Structures concerned with yolk
absorption in the dogfish.

Dr. Eric LOEWENSTEIN: Demonstration of fluoro-
photometer, and discussion of fluorometric methods of
determining biological substances.

Dr. LAURENCE IrvING, Dr. P. F. SCHOLANDER, MR. C.
Lioyp Cuarr, Mr. GrorGe Epwarps, Mr. Nrets Hav-
GAARD: Section A: Sensitive volumetric apparatus de-
vised by Dr. P. F. 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 O, 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

[ Vor. 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 28, 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 belongs: 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 describes the dissection of
the usual laboratory types in clear language but
also discusses classification, phyletic origins and
ontogenesis. All anatomical terms are italicised
and the author holds a happy balance between the
English and Latin tongues. The absence of illus-
trations and the paper 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 paper and clearer printing 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 Microscopie 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 taxonomy 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:

ProfeSSOES) ....cssccssesscssessassdancceesasostessosceccoumeeeremeteeeeeeaee 66
Associate Professors .. 23
Assistant Professors ... 42
IMStLUCtOrsimeeccseceereeee 35
Research Associates . tl
IASSISGANLS) tecessceeesaseres 55
PQMWOWS. ieescessesvicecsesssastadexvsasevasen tre eee 20
Graduate Students (not listed elsewhere).. OU
Medical! Students) <.i.s2-cccssecse-cseoessesseereeeneeee a
Undergraduate Students ... ee
Preparatory Students .............:0 el
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 been:

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 in the Hole turns to run
from Buzzards Bay to Nitevard Sound:

Date -M. P.M.
PNGRAGEE SIL eececdeocects L1s39) eae
Septembers lee 12:24 12:42
September Zee 1:25 lh:43
September oy 22s [aay 4339)
September 4 3:14 3:30
September Oye ees 4:01 4:18
September 6 ............ 4:42 5:00
September y/aeee ee DE23 ORAZ
September 8 ............ 6:05 6:25
September 9)..21...... 6:45 7:09

Aucust 30, 1941 |

Wald, COWMLIACAMUNE; Naar

187

ITEMS OF

The invertebrate course of the Marine Biologi-
cal Laboratory is holding its last session this
morning.

Dr. Corin M. MacLeop, 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. ReyNnotps 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. WittiAm 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. Exso S. BarGcHoorn, JR. of Harvard Uni-
versity has been appointed instructor in the biol-
ogy department of Amherst College.

Mr. Stipney M. Ponp, 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.

INTEREST

Recent visitors at the Laboratory include Drs.
H. J. Muller, Gertrude Gottschall, Oscar Bodan-
sky, Elvira de Liee, Benjamin C. Gruenberg,
Hans Gaffron, R. Leuchtenberger, 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 15 for Camp Devens; H. Duncan Rol-
lason, Jr., left last week for Connecticut to take
his physical examination.

Mr. Ermer Hiccins, chief of the Division of
Inquiry 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. GerALp 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. Leonarp I. Karzin, who worked at the
Marine Biological Laboratory last year, is at
present 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 Marcaret 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.

THE COLLECTING NET

[ Vor. XVI, No. 147

IN MEMORY OF DECEASED MEMBERS OF THE CORPORATION OF THE
MARINE BIOLOGICAL LABORATORY

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
member distinguished for his accomplishment in
research and even more for his quality as a man.
Tennent was an investigator who proceeded with-
out haste, yet 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 known to many investigators in other
lines, because i ennent was the most modest and
retiring of men. He had none of the flare for
self-advertisement that carries some men so far
on a modicum of worth. His every publication
was marked not only by the critical nature of his
observations and experiments but also by the
meticulous care with 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 advance 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 did his research. Tennent and I were
graduate students together 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

Memorials Adopted at the Annual Meeting of the Corporation, August 12, 1941

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 where he might have had more “‘disci-
ples”. He trained many 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

Aucust 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

Meals, COMURKINING, INS

[ Vor. 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 were 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 im-
provement 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 they were formed,

but his conclusions were always based on a clear-
sighted and critical consideration of evidence, in
the collection of which 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
eift 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.

IRS, Wan ine

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 Harbor Symposium, 1941, forthcom-
ing). Such a situation explains the existence of
symmetric arrangements of R-groups such as are
known to be present in insulin, for example It
also fits with the existence of precursors such as
Svedberg’s prolactalbumin and several other pro-
tein intermediates, which 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 may
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 in general provide 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 zine
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 Euglena can live in a concentration of
10-8 M copper sulfate as long as food lasts, but
dies in a few minutes when the concentration
reaches 10-7 M (Seybold, Biol. Zebtralb., 47, 102,
1927) ; that Amoeba proteus cannot survive when
the concentration of CuCl. 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

Aucust 30, 1941 ]

THE COLLECTING NET

191

fundamental superspecificity of biologically active
structure systems. For the specificity of individ-
ual native protein units, conjoined with specificity
of interlinking to give the superspecificity 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 by 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
CuCly 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)», 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

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 de
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 inteuded 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 been
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 possibly 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.)

INVERTEBRATE CLASS NOTES
(Continued from page 184)

—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 grabbed
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 Batchelor

192 THE COLLECTING NET

[ Vor. XVI, No. 147

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Aucust 30, 1941 | THE COLLECTING NET 193

Turtox Quiz Sheet Subjects for Every Teaching Need

The Turtox series of Key Cards and Quiz Sheets now comprises the most complete
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194 THE COLLECTING NET [ Vor. XVI, No. 147

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Aucust 30, 1941 |

Designed to give all types of illumina-
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The triple lens condensing system is in
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THE COLLECTING NET

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195

THE COLLECTING NET

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[ Vor. XVI, No. 147

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