Abstracts
REZNEAT M. DARNELL, el al., Marquette University.
Symposium: Animal Nutrition.
Recent years have seen a reawakening of interest
in the field of animal nutrition. Armed with calorimeters, isotopic tracers, computers, and a portfolio of chemical tools, nutritional ecologists are
finding themselves deeply involved with bioenergetics, incorporation and turnover rates, and environmental biochemistry. In these areas the gap
between the ecologist, the physiologist, and biochemist has become blurred, and interdisciplinary
dialogue is likely to prove quite rewarding. The
area of food chains, trophic levels, and community
metabolism, although more distinctly ecological, is
strongly rooted in the function of the individual
organism and may be viewed as an extension of
areas of common interest. The present symposium
on animal nutrition is an attempt to develop more
formal communication between ecology and the
related disciplines, to summarize the present state
of knowledge in the various areas, and to focus attention on currently active fields of investigation.
WILLIAM THURMOND, California State Polytechnic College, San Luis Obispo, California
93401.
Intermedin-like activity by the pars distalis of the
salamander Ambystoma tigrinum.
Portions of the pars distalis from adult tiger salamanders were homoplastically transplanted to subcutaneous tissue of "albino" larvae previously hypophysectomized at the tailbud stage. The majority
of the transplants from areas adjacent the pars intermedia caused a sustained dispersion of melanin
granules in the host's melanophores surrounding
the graft. On the other hand, most grafts from
areas away from the distalis-intermedia junction
evoked darkening which disappeared after 4-8 days
and then reappeared within 1-5 days and persisted
for the duration of the observation period (16-48
days).
Thyroids were placed adjacent to the hypothalamus and pars intermedia of completely and partially hypophysectomized adults respectively for one
week then subdivided and transplanted to larvae
to serve as controls to determine possible diffusion
of melanophor-stimulating material into the pars
distalis. Hypothalamic conditioned thyroid grafts
evoked pigment responses for 2-3 hours whereas
thyroid grafts with intermedin caused darkening in
host larvae for 24 to 36 hours.
Control grafts of pars intermedia evoked maximum dispersion of melanin granules in all the
host's melanophores and within a few weeks hyperpigmentation was clearly evident. A majority of
grafts of pars nervosa, on the other hand, evoked
localized pigment responses for only 2-10 days.
(This research was conducted at the Jackson Hole
Biological Research Station, Moran, Wyoming.)
8
S. MATSUO, A. VITUMS, J. R. KING and D. S.
FARNER, Washington State University and University of Washington.
Cytological changes in the adenohypophysis of the
male White-crowned Sparrow, Zonotrichia leucophrys gambelii, during its annual cycle.
Cytology of the adenohypophysis, especially its
pars distalis has been investigated by a variety of
cytological and cytochemical methods. The hypophyses were obtained from birds in different periods of their annual cycle.
The pars distalis has a distinct tinctorial differentiation between the amphophils located exclusively in the cephalic lobe and the acidophils which
occupy only the caudal lobe. The amphophils appear blue-violet and the acidophils red or orange
after applying tetrachrome staining (Matsuo, 1954).
The amphophils are PAS positive (purple cells).
The acidophils usually are PAS negative but sometimes a slight PAS positive reaction has been observed. Many acidophils change their tinctorial
properties and become amphophilic and smaller in
size at the end of April and May.
Large basophils appear light-green after tetrachrome staining and are PAS positive (red cells).
These cells are observed throughout both lobes
from the middle of February and gradually increase
in number reaching a maximum at the end of
April and May. They suddenly decrease in number and size at the middle of June, and almost
completely disappear from September through
January. The intensity of the PAS reaction of
these cells is also maximal from the end of April
through May. Our findings correlate well with the
annual reproductive cycle when gonads reach their
maximal weight in May (Farner, 1964) and also
when gonadotropic potency of the hypophysis is
the greatest (King et al., 1966). This suggests that
these cells may be concerned with the gonadotropic
functions. (Supported by Grant 5R01-HEO7240
NEUA from the National Institutes of Health.)
ROGER A. HOFFMAN, Colgate University.
The pituitary gland of the chinchilla (Chinchilla
lanigera).
The pituitary gland of the chinchilla lies in a
shallow sella turcica, and is heavily invested by a
tough tectorial membrane. It is flattened dorsoventrally, and is elongated along the antero-posterior axis with sample dimensions of about 5.3 X
3.4 mm. The short free stalk is attached to the anterior end of the pituitary gland and courses posteriorly along the top of the anterior lobe to end in
the pars nervosa which occupies the posterior onequarter of the gland.
Along the ventral and lateral surfaces of the infundibular stalk, a prominent pars intermedia contains very obvious cells of two types. The darkcells which are frequently spindle-shaped and
border on the pars nervosa or upon the residual
187
188
AMERICAN SOCIETY OF ZOOLOGISTS
cleft are PAS, AF and alcian blue positive. These
cells stain prominently with AF+ without prior oxidation whereas the typical AF basophils remain
unstained unless first oxidized. Frequently, the cells
of the pars intermedia can be seen invading the
pars nervosa and in such cases, no basement membrane can lie demonstrated between the two tissues.
Neurosecretory granules are frequently found in direct contact with the cells of the pars intermedia.
Cold exposure, castration, starvation or lack of
water lead to colloid-cysts within both the pars anterior and intermedia. On the basis of easy and
obvious cell differentiation in the pars intermedia,
this animal may prove of value in studies concerned with the function of the mammalian pars
intermedia.
10
JERRY W. DUSSEAU, ALBERT H. MEIER, and
ETHELYN LEHAYE, Louisiana State University.
Seasonal and diurnal levels of plasma adrenocortical hormone in the White-fhroated Sparrow.
The levels of plasma adrenocortical hormone in
the White-throated Sparrow, Zonotrichia albicollis,
were measured fluorimetrically at three seasons of
the year. Photosensitive birds were sacrificed in
February and May, and photorefractory birds were
sacrificed in August. The blood samples were collected every six hours of the day, beginning at sunrise, at each of the three seasons. A marked depression of the plasma adrenocortical hormone was observed during the photorefractory period, indicating that the interrenal tissue of the White-throated
Sparrow is less active at this time of year than it
is during the photosensitive period. A diurnal
rhythm of the plasma levels of adrenocortical hormone was observed in the photosensitive birds.
However, no rhythm was found in the photorefractory birds tested in August. Both the seasonal and
diurnal levels of the adrenocortical hormone may
be correlated with certain events of the annual
cycle, such as migratory behavior and its accompanying physiologic events. (Supported by Grant
GE-3679 from the N.S.F.)
11
ROBERT ORTMAN, City College of New York.
The Napoleon Weaver and the Orange Weaver in
the weaver finch test.
Ortman (Amer. Zool. 6: 518, 1966) reported on
experiences with the Paradise Whydah in the
weaver finch test for luteinizing hormone (LH)
and human chorionic gonadotropin (HCG). In
these experiences the Paradise Whydah failed to
show the desired reactivity (except for one bird).
It was thought valuable, therefore, to investigate
the reactivity of other weaver birds: we have begun
these investigations with relatively high doses. A
detailed study of the reactivity and the hormone
requirements of the Napoleon (Eiiplectes afra) and
the Orange (Pyromelana jransiscana) weaver is being made and shall be reported on. At the time of
writing, some preliminary results are available.
The Napoeon W.F. (7 birds) has given positive
reactions with total doses (administered as a single
daily dose for 2 days) ranging from 50 to 384 IU
of HCG; one Napoleon W.F. which was injected
with 192 IU of HCG did not give a positive reaction. Two Orange W.F. gave positive reactions
with 384 IU of HCG; one Orange weaver failed to
react at 384 IU. Moreover, four Orange weavers
failed to react at dose levels between 50 and 150
IU of HCG. Control birds which were injected
with sterile water gave no positive reactions. (Supported by Grant AM 07204-03 from the U.S.P.H.S.
and by Biology Department funds.)
12
HENRY C. BROWNING, University of Texas,
Dental Branch.
Induction of LTH release in mice by norethynodrel-mestranol.
Intraocular ovarian isografts.in castrated female
mice indicate pituitary release of LH by forming
corpora lutea and of LTH release by development
of hyperemia in these corpora. Such grafted BALB/
c or CBA mice, in groups of 10 to 14, showed normal vaginal cycles. In BALB/c mice, hyperemic
corpora developed in 95 of 108 cycles; the duration
of hyperemia was 1.9 ± 0.18 clays. Such hyperemic
corpora appeared in only 33 of 83 cycles of CBA
mice; the duration was 1.4 ± 0.10 days. Similar
groups of each strain received 98.5 jug of norethynodrel with 1.5 /ig of mestranol (Enovid, Searle), subcutaneously in saline suspension, daily for 3 months.
All mice ceased to show vaginal cycles within 5
days and thereafter remained in metestrus. The
ovarian grafts also ceased to show cyclic changes
and maintained vesicular follicles only. At the end
of 2 months, each animal in the treated and untreated groups received 3 consecutive daily doses of
5 fig of LH intraperitoneally in saline. Cycles and
duration of hyperemic corpora were unaffected in
untreated mice. In treated BALB/c mice, hyperemic
corpora developed; the duration of hyperemia was
6.8 ± 0.87 days. In treated CBA mice there was a
similar response but the duration of hyperemia
was 3.3 ± 0-45 days. It appears that norethynodrelmestranol does not alfect FSH release in treated female mice but does inhibit that of LH; at the
same time, LTH release is stimulated. (Supported
by USPHS Research Grant CA 02880.)
13
STUART RUDIKOFF and D. A. OLEWINE.
Georgia Southern College.
Ligation of the utero-cervical junction of pregnant
rats and the disruption of pregnancy.
Recently, much attention has been directed toward investigating the manner by which an intrauterine suture prevents pregnancy in the rat. To
be effective such sutures must extend into the lumen of the uterus since failure to do so results in
implantation. We have also found that sutures
placed in the cervix of pregnant rats, where implantation does not normally occur, interferes with
pregnancy but does not totally prevent it. In the
present study, the utero-cervical junction was ligated during pre-implantation (day 4 of pregnancy)
or during post-implantation (day 7 of pregnancy).
All animals were sacrificed on day 14 of pregnancy.
At this time many of the uterine horns ligated on
ABSTRACTS
day 4 of pregnancy were free of implantations.
However, the number of implantations in uteri ligated after implantation were similar to the number found in uteri from sham-operated and control animals. These findings indicate that ligation
of the utero-cervical junction on day 4 of pregnancy is an effective inhibitor of implantation.
14
ROBERT L. HAZELWOOD and J. R. KIMMEL.
University of Houston and Kansas University
Medical School.
Effect of avian insulin on glucose uptake by diabetic rat diaphragm. (Introduced by N. W.
Klein).
Crystalline insulin was prepared from fresh frozen chicken pancreas by homogenization, acid alcohol extraction, acetone powder preparation, purification by gel filtration and subsequent chromatography. (Fed. Proc. 25:761, 1966). Amino acid
composition, as based upon examination of 20 and
40 hour hydrolysates, indicated 51 amino acid residues/mole of protein; Che theoretical values of individual amino acids agreed well with the determined values.
The avian product was assayed along with
equivalent amounts of crystalline beef insulin employing an in vitro system of hemidiaphragms
either from fasted normal or from diabetic rats.
Glucose uptake and glycogenesis were measured
after a one hour incubation in a metabolic shaker
at 38°C. In other studies, the influence of preincubation of different insulins with diabetic rat plasma
was determined by employing the above techniques.
The results obtained indicated that the avian hormone was less potent in promoting glycogen deposition in the normal diaphragm (41 vs 49% at 0.1 pi
cc and 50 vs 61% at 0.2 fi/cc) and was markedly
less potent in this respect when tested on diabetic
diaphragm (21 vs 40% at 0.1 ^/cc and 26 vs 55%
at 0.2 p./cc). Diabetic rat plasma inhibited the glucose uptake effect of both bovine and avian insulin; however, glucose uptake with avian insulin was
affected to a greater extent (25%).
It appears that avian insulin is inhibited by the
presence of diabetic rat plasma, or when acting directly on the diabetic rat diaphragm, is less potent
than bovine insulin in encouraging glucose uptake
and glycogenesis. (Supported by NSF: GB-2752 and
NIH: AM-O9O72-O2).
15
ELDEX W. MARTIN and LEE E. FABER, Bowling Green State University.
The effects of ultrasonic sound on auditory pathway potentials in the laboratory rat.
The response of wild rats to ultrasonic sound has
been the basis for one method of rat control. This
study was conducted to provide physiological verification that auditory transmission of sound in the
high frequency range was possible in the Tat. Macroelectrode implants in the inferior colliculus were
used to monitor electrical activity which was displayed on an oscilloscope. Anesthetized laboratory
rats were exposed to clicks, white noise, and pure
tones of frequencies from 5 to 100 kilohertz. The
189
clicks which evoked spike potentials and the white
noise which produced increases in background activity were used to verify implantation in an auditory tract. Pure tones caused a decrease in background activity.
The upper limit of pure tone frequencies which
elicited responses varied with the individual rat
and the level of anesthesia. Tones of 40 to 50 kilohertz produced the most consistent results as an
upper limit. It was concluded that the inferior colliculus of the rat, and hence the auditory nerve, is
capable of transmitting neural activity generated
by ultrasonic sound. (Supported by Grant 222177
from The Peavey Company, Minneapolis, Minn.)
16
VALERIE ERNST, University of Louisville.
The structure and function of the proboscis retractor muscle of the sipunculid, Golfmgia gouldu.
The electrophysiology of the proboscis retractor
muscles of the sipunculid, Golfingia gouldii, have
been extensively studied by Prosser and co-workers
(J. Cell. Comp. Physiol., 38, 299, 1951). The muscles
are nonstriated and can respond both phasically
and tonically according to innervation suggested by
Prosser, et al. The fibers are about 1.0 mm long
and 5 p wide and each appears to be surrounded
by a network of collagen. The muscle is extremely
extensible and can easily be stretched to several
times its resting length. Histological investigations
reveal a "basket-weave" type of arrangement of
straight and zigzag muscle fibers which, it is believed, facilitates this hyperextensibility. The
muscles are thought to be dually innervated (Prosser, et al., J. Cell. Comp., 54, 129, 1959) and neurons containing neurosecretory granules or synaptic
vesicles have been observed in the electron microscope.
These observations suggest that histochemical investigations at the light and electron microscope
levels could be used to distinguish between those
fibers innervated by acetylcholine from synaptic
vesicles and those innervated by neurons containing neurosecretory granules.
17
YORIMI MATSUMOTO and B. C. ABBOTT,
University of Illinois.
Folding non-striated fibers of the Golfingia gouldii
proboscis retractor muscle.
The proboscis of Golfingia gouldii functionally
extends approximately to a maximum of 50% of
the animal length and can be completely inverted
into the body. The retraction of the proboscis is
achieved by non-striated muscles consisting of fiber
units 5 p. X 1000 p. Folding of these fibers is indicated histologically (Prosser, et al., J. Cell. Comp.
Physiol., 38, 299, 1951). The fiber orientation is also
determined by the birefringent nature of the
muscle. The birefringent band width is correlated
to the active tension length relation to establish
fiber folding.
18
THEODORE L. JAHN, University of California,
Los Angeles.
190
AMERICAN SOCIETY OF ZOOLOGISTS
A possible mechanism for the effect of electrical
potentials on apatite formation in bone.
It is assumed that when an electrical potential is
applied to bone, the organic matrix and apatite
constituents behave as amphoteric ion exchangers.
On this basis application of a continuous potential
of external origin should cause accumulation of
calcium and phosphate ions at the cathode and of
sodium and chloride ions at the anode. The resulting distribution of these ions can contribute to the
formation of apatite at the cathode and to possible
resorption at the anode, and the increase in mesenchymal cells and osteoblasts observed at the cathode might be caused by galvanotaxis, similar to
that of Amoeba proteus.
The surface of the apatite crystal bears fixed
positive and negative charges (Ca and phosphate
ions) with which counterions are associated. Chloride and phosphate ions compete for the calcium
sites, and sodium and calcium ions compete for the
phosphate sites. These association-dissociation equilibria can be modified by application of an electrical potential, even if alternating slowly, as in the
naturally occurring deformation effect. The deformation potential causes the association of phosphate to be accelerated and the association of calcium to be decelerated. However, the acceleration of
phosphate capture is several times greater than the
deceleration of calcium capture; therefore, the net
effect should be an increase in the formation of
apatite. This electrical effect makes the usual assumption of ionic pumps unnecessary. The dependence of association-dissociation equilibria on deformation potentials provides a mechanism for Wolff's
law. (Supported by Grants NIH 6462, 8611; NSF
GB 1589, 5573; and contract NONR 4756.)
19
WAI-MAN AU, Chu Hai College, and JOSEPH
C. C. HWANG, Chung Chi College, Chinese University of Hong Kong.
Transfer function of a crab mechanoreceptor.
The input and output relationship of stretch receptors in the meropodites of the pereiopods of the
crab Cancer magister was analysed. The input in
the form of angular changes of the mero-carpopodite joint was measured in degrees. The output in
the form of pattern of impulses from a group of
bipolar sensory neurons in the meropodite was then
studied. The relationship between the input and
output of this sensory system could be expressed as
a transfer function to be determined by way of
Laplace Transformation. The transfer function
proposed was as follows:
Eo(s)
with
F (s) =
E,(s)
Tos
T being the duration of movement, and X being
the angle through which the joint has been moved.
Assuming initial condition of eo| •== O,
|t=o
1
.
To
__
f
T,
eo = CkT (e — 1) e + CX when t > T
The effects of moving the joint at different rates
on the pattern of impulses evoked were analysed.
The respective values of C, T o . k, T, T, and X were
recorded and calculated. Movements at three different rates of 10°/sec., 16°/sec., and 24°/sec. were
effected. For movement at the rate of 24°/sec. in
one preparation, k = 24°/sec, C = 0.49, T o = 2.04
sec., r = 1.2 sec, Tr=2.2 sec. and X = 10°. In the
three cases analysed, a very good correlation was
obtained in each case between the theoretical curve
predicted and that plotted with the actual data,
thereby justifying the adoption of the above equation.
20
ROGER D. FARLEY, JAMES F. CASE and K. D.
ROEDER, Tufts University and University of
California, Santa Barbara.
The neural control of ventilation in Periplaneta
americana (L.).
When agitated or exposed to excess CO2 a cockroach ventilates its tracheae with dorso-ventral abdominal movements at a frequency of 1-3/sec. The
expiratory phase is synchronized with spike bursts
in fibers lying within the abdominal cord, while
inspiration depends upon tissue elasticity. Analysis
of impulse direction in the ventral nerve cord,
along with cord sectioning experiments, showed
that the metathoracic ganglion is usually the pacemaker center for these abdominal movements. Impulses travel from the metathoracic to the terminal
(6th abdominal) ganglion along internunciary fibers
with a conduction velocity of approximately 3 m./
sec.
Rhythmic motor activity similar to that during
normal ventilation was recorded from isolated nerve
cord preparations, and sometimes from single isolated abdominal ganglia. In more intact preparations stretching or compressing the abdomen of a
headless insect during ventilation delayed the arrival of the immediately following motor burst.
This suggests that output from abdominal proprioceptors modulates the on-going pacemaker
rhythm, and that feedback coordinates the two
oscillating systems—the abdominal musculature
and the respiratory pacemaker. (Supported by NIH
postdoctoral grant, USPHS Grant NB-04372 and
USPHS Training Grant Tl AI 32.)
21
ROBERT G. SHERMAN and RALPH A. PAX,
C = C2 and T =
Ts
e, (t) — kt when t =£ T
e, (t) = X when t > T
Michigan State University.
A physiological and morphological study of a spider
heart.
One class of arthropods in which cardiovascular
physiology has been little studied is the class
Arachnida. To obtain further information about
this class, we have undertaken a study of the heart
191
ABSTRACTS
of the spider, Geolycosa missouriensis.
This heart is a tubular structure approximately
6 mm in length, situated mid-dorsally in the anterior two-thirds of the abdomen. A cord of tissue,
similar in location and gross appearance to the cardiac ganglion of Limulus, extends the length of the
heart. Silver staining of this cord shows that it contains a number of cell bodies from which arise typical neural-like processes. The mean rate of beating
as calculated from electrocardiograms recorded
from nine intact spiders was 145 beats a minute
(SD 25.6 beats a minute). Hearts removed from
spiders continue beating with little change in rate
for as long as four hours but the rate is markedly
less than that in intact animals. The mean rate for
nine isolated hearts was 60 beats a minute (SD
13.3 beats a minute). The pattern of electrical activity recorded from both intact and isolated hearts
is of the oscillatory type characteristic of neurogenic hearts. The pattern consists of a series of fast
waves, each series characterized by an initial large
wave followed by several smaller ones.
Both the histological findings and the nature of
the electrical activity recorded indicate a possible
neurogenic origin for the heart beat in this spider.
(Supported by Fellowship 1-F1-GM32.OO7-O1 from
the U.S.P.H.S.)
22
MOON JAE PAR and BERNARD C. ABBOTT,
University of Illinois.
Contractile properties of toadfish sonic muscle.
Mechanical and electrical properties of isolated
small bundle of sonic muscle from toadfish swimbladder was studied. Attachments were made to
tendon on one end and to a piece of swimbladder
wall on the other. It was stimulated massively.
Adequacy of the bathing solution was attested by
the prolonged survival of the tissue.
Resting potential at 20°C averaged 74.2 mv and
action potential 88 mv. Isometric tension began
during the action potential, reached the peak in
about 4 msec and complete relaxation took place
in the next 4 msec. Fused tetani were 2not obtainable
and peak twitch tension of 70 gm/cm was maximal
at 6°C. The maximum rate of tension rise showed
the same kind of temperature dependence as peak
tension.
The muscle at 20°C manifested remarkable dependence of its twitch tension on stimulation frequency and also post-repetetive-stimulation potentiation. At optimum frequency, 4-5 times potentiation of resting peak tension was observed.
In order to ascertain whether the extremely fast
contraction is due to rapid excitation-contraction
coupling process (Markedly well developed sarcoplasmic reticulum in this muscle was reported
(Fawcett & Revel, J.B.B. Cytl. 10:89, 1961)) or to the
mechanism residing in the contractile element, tension development in glycerol-extracted fiber was
studied. Extracted fibers did not respond to wide
range of ATP and Ca concentrations. However, when
pretreated with strong surface active agent, tension
of about 1/100 of psoas fiber of comparable size
was developed very rapidly.
It was tentatively concluded that the rapidity resides in both coupling process and unique contrac-
tile property. (Supported by the Grant NONR
395-02 from the U.S. Navy.)
23
THOMAS F. DE CARO, PMC Colleges, Chester,
Pa.
Calcium movements and contractility in rabbit atrium.
Calcium in rabbit atrium can be divided into at
least two components by employing twitch-tension
studies coupled with kinetic studies with Ca15. Left
atria removed from freshly dissected hearts were
cannulated and inflated with a gas mixture containing 95% O2 and 5% CO3. They were depleted
of calcium by immersion in a Krebs solution containing low calcium (0.2 mM), and then treated in
one of two ways: either soaked for 3 minutes
(group I.) or 2 hours (group IIa) in a Krebs solution containing 5 mM CaCI2. When group I . atria
were stimulated at a frequency of once per minute
in a Krebs solution containing low calcium (0.2
mM), twitch-tension declined rapidly, whereas the
half-time of decline in group II, atria was 70 minutes.
In addition, atria employed in the isotope study
were also depleted of calcium and then soaked for
either 3 minutes (group Ib) or 62 hours (group IIb)
in a solution containing Ca* . They were then
washed-out in successive changes of a non-isotopic
solution. Calcium exchange in group Ib was rapid,
but the half-time of exchange in group IIb was 80
minutes.
These findings suggest the presence of at least
two calcium moities involved in contraction in
atria. One is probably superficially bound to extracellular sites, the other is most likely stored within
the cells. (Supported by research grants NBO440904
from U.S.P.H.S. and 6F2HE3099301A1 from N.I.H.)
24
DOROTHEA CASKEY MANGUM, Baylor University, Waco, Texas.
An analysis of the burrowing behavior of a sea anemone from the Gulf of California.
The physiological and morphological properties
of the nervous and muscular systems of the tropical
burrowing anemone, Phyllactis concinnata, were
compared with those of well-studied forms, such as
Metridium, Calliactis and Stomphia. The burrowing behavior of this anemone is described and compared with other purposeful and coordinated movements of anemones. Burrowing is accomplished by
the inflation and contraction of the pedal disc as
the column shortens and elongates. This results in
progressive attachment and loosening of the pedal
disc. Movements associated with burrowing appear
similar to other aspects of anemone behavior such
as walking, feeding, and the slow inherent movements which also require the parietal and circular
muscle contraction sequence. The most unique feature of burrowing is the consistent and predictable
quality of the rhythmic contractions, which may be
maintained for as long as 6 hours. (Supported in
part by NSF Grant GB-3745 and by Pre-Doctoral
Training Grant, USPHS #2 T 1-GM 441-04.)
192
AMERICAN SOCIETY OF ZOOLOGISTS
25
HYMAX GUTHWIN and DANIEL FRIEDMAN,
Hunter College of the City University of New
York, New York.
The fate of Hydra littoralis in the gastrovascular
cavity of Hydra littoralis.
The ingestion of Hydra by Pelomyxa and of Pelomyxa by Hydra with subsequent egestion, alive, of
each by the other, was reported before the Society
of Animal Behavior, December 1966. The ingestion
o£ Hydra by Hydra is reported here. In 1959, W. F.
Loomis described and pictured eversion and partial
cannibalism in Hydra under the influence of glutathione. Neither complete ingestion nor sequelae
were suggested. We have observed, during feeding
with Anemia salinis, both partial and complete ingestion of Hydra by Hydra. When only tentacles
or tentacles and proximal body were ingested, the
captured Hydra was released within half an hour.
When Hydra was totally ingested, the tentacular
end of ingested Hydra was directed towards the
basal end of ingesting Hydra since the tentacular
ends of both were directed at the brine shrimp.
Ciliated protozoa ingested by each along with the
brine shrimp, remained intact long after the brine
shrimp had been reduced. These protozoa were inactivated and lost morphological integrity sooner
in the ingesting Hydra than in the ingested Hydra.
The ingested Hydra was expelled intact after a sojourn of at least two hours in the digestive cavity
of ingesting Hydra. Subsequently, each Hydra
egested the remainder of its brine shrimp and ciliate meal. The egested Hydra, fed Artemia, successfully immobilized and ingested the brine shrimp.
26
SIDNEY C. HSIAO and S. ARTHUR REED, University of Hawaii.
Separation and partial characterization of alkaline
phosphatase from Fungia scutaria, a fungid coral.
Alkaline phosphatase was separated from young
Fungia scutaria, a fungian Medreporaria, by different methods. The soft tissues were separated by
autolysis under toluene, trypsin digestion as a whole
or after pulverization, and manual separation followed by homogenizing in a Virtis homogenizer or
blending in a Waring blendor. The autolysate, digest, homogenate and brie were separately treated
with methods reported in the literature on alkaline
phosphatase purification: n-butanol, ether, ammonium sulfate, ethanol, chloroform, dialysis and
chromatography. Comparison of the total enzyme
obtained per unit of material used and the degree
of enzyme concentration achieved showed that
manual separation combined with butanol and ammonium sulfate treatments gave the best result.
The effects of temperature, pH, dialysis and lyophilization on the activity of this enzyme were examined. The alkaline phosphatase of fungid coral
exhibits different hydrolytic action on different substrates. From these studies the temperature characteristics, optimum pH and the most easily hydrolyzed substrates were determined.
27
LOUISE RUSSERT KRAEMER, University of Arkansas.
The mantle flap in three species of the genus Lampsilis (Pelecypoda). (Motion picture)
The mantle flaps are a permanent anatomical
feature of the mature female, as an extension of the
inner lobe of the mantle edge just anteroventrad to
the branchial siphons. Among the flaps of three
species investigated, structural similarities (presence
of eyespot, innervation by branches of siphonal
nerves from the fused visceral ganglion) and differences were found.
Flap movements are all comprised of: (a) paired
pulses initiated as contractions at the tail base and
moving toward the eyespot ends of the flaps, and
(b) a recovery phase in which the flaps assume their
former position, with tails floating horizontally.
Flapping behavior is not confined to flap movements but involves the coordinated function of foot,
marsupium, valves and siphons to such an extent
that the supposed normal relationships between
body and shell are much altered. Flapping behavior
involves different behavioral complexes as well as
different relevant stimulus modalities for different
species (especially light for L. ventricosa and water
waves and jarring of substrate for L. siliquoidea).
Flap movements per se accompany spawning of
glochidia in all species in which the movements
have been observed. Evidence in support of this
statement includes: (a) flaps occur only in mature
female specimens, although juveniles and males
have flap rudiments; (b) flap movements have been
seen only in gravid, never in non-gravid females:
(c) flap movements have been seen in association
with gradual emptying of the ovisacs, and with
shedding of conglutinants; and (d) flap movements
have not been observed after shedding of glochidia.
28
OZRO B. WISWELL and HENRY C. BROWNING,
University of Texas Dental Branch, Houston.
Reproduction of the giant hermaphroditic snail
Slrophocheilus oblongus. (Motion picture)
A colony of the terrestrial gastropod, Strophocheilus oblongus, has been established. The adults
weigh between 70-100 gms., and the foot when extended is 12-15 cm. long. An apparently adequate
diet has been established in that no adult animal
has died and mating has been observed. Two hundred and forty-seven eggs have been produced in
57 clutches of one to six eggs. Our observations
concerning timed matings suggest an egg maturation time of 19.7 days with the subsequent average
incubation time of 53 days (a few as short as 34
days and others as long as 70 days) at a relatively
constant temperature of 21.1° ± I°C.
This color motion picture film demonstrates estivation, phytophagic activity, mating, egg clutches
and hatching of eggs. (Supported by General Research Support Grant 5-S01-FR-5344-05.)
29
ALAN E. ORGAN, EUGENE C. BOVEE, DUNCAN WIGG, and JAMES R. FONSECA, Univer-
193
ABSTRACTS
sity of California, Los Angeles.
The mechanism of the nephridial apparatus of
Paramecium multimicronucleatum. I. Expulsion
of water from the vesicle. (Motion picture)
Recent analysis of the mechanism of the nephridial apparatus of Paramecium multimicronucleatum
by high speed cinematography (300 p.p.s. at 250
X) confirms the observations by electronmicroscopy
(Schneider, /. ProtozooL, 7: 75-90: 1960) that the
vesicle is invaginated by adjacent cytoplasm, once
the pore is opened, and is emptied by collapsing
under pressure from that cytoplasm, aided perhaps
by pressure of the fibrils which anchor the ampullae to the excretory canal. There is no indication
of active contraction of the vesicle or its membrane.
There is no permanent pore to the vesicle. It is
closed by a sealing of the ruptured membrane
where it is in contact with the pellicular excretory
canal. At onset of expulsion the membrane across
the basal opening of the excretory canal is ripped
along one semicircular portion of the excretory
pore, and is driven up against the opposite wall as
a flap while the water rushes out. This flap drops
back as pressure is reduced; and a constriction of
the vesicular and cell membranes at the base of the
excretory canal reseals the opening. (Supported in
part by USPHS Grant 6462.)
30
BERNICE F. PIERSON, ROBERT GIERKE and
ALBERT L. FISHER. Montgomery Junior College, Rockville, Maryland.
Clarification of the taxonomic identification of Euplotes eurystomus Kahl and Euplotes aediculatus
Pierson.
Morphologic studies were made on specimens
stained with picro-indigo-carmine, impregnated
with silver (Chatton-Lwoff as modified by Corliss
1953) and in the living state. The living animals
were observed with a prototype Francon-Yamamoto differential polarizing interference microscope.
The number of rows of lateral-dorsal kineties was
constant: eight in Euplotes aediculatus and ten in
Euplotes eurystomus.
The peristomal cavity of E. eurystomus transects
the anterior-posterior axis of the body while that
of E. aediculatus roughly parallels it. The anal
cirri are inscribed along their posterior border in a
V-shaped arangement in both species but the distance between the posterior edge of the buccal
cavity is much greater in E. eurystomus.
The macronucleus from a ventral view is typically "3"-shaped in E. eurystomus while in E. aediculatus it is the mirror-image of the letter "C."
Study of peristomal lip movements and positions
in the living state gave further confirmation of the
validity of the above structures as bases for the
identification of these two species.
The peristomal edges of both species opposite the
line of adoral membranelles were seen in living
animals to be cast as a thin undulating structure
suggestive of a membrane. This is apparently the
first observation of an undulating membrane in
this group of ciliates.
The present investigation was an extension of
work reported by the senior author at the Second
International Conference on Protozoology, London,
August 1965.
31
HARLEY P. BROWN, University of Oklahoma.
Psephenids (Coleoptera: Dryopoidea) parasitized
by eulophid and darapriid wasps (Hymenoptera:
Chalcidoidea and Proctotrupoidea).
In the region of Sabinas Hidalgo, Nuevo Leon,
Mexico, Psephenus texanus (a new species being
described elsewhere) is heavily parasitized in the
prepupal and pupal stages by wasps representing
two unrelated families. From 7 to 39 adults of
Psepheuivorous (Eulophidae: new genus and species being described by B. D. Burks) may develop
within a single host. Only a single individual of
Trichopria (Darapriidae: new species) parasitizes a
host. Psephenivorus larvae usually consume the entire contents of the host. A Trichopria larva does
not consume all; it leaves the posterior portion of
the victim, as well as a sac of feces. The life cycle
of Psephenivorus may be completed within 21 days;
that of Trichopria takes longer. Both spend their
larval and pupal stages within the host, emerging
as adults. The host is invariably killed, but its bodyprovides excellent protection for the developing
parasites, the carapace of the last larval skin forming a turtle-like covering for the pupal host. In
the region studied, at least 50% of the pupating
Psephenus were parasitized by Psephenivorus, and
at least 25% by Trichopria. The host occurs in such
Texas streams as the Nueces and Rio Frio. Both
species of wasp may be expected there. The author
has found no parasites in association with psephenids in other parts of the United States, nor any
published records of parasites upon dryopoids elsewhere. (Supported by the University of Oklahoma
Faculty Research Fund.)
32
KENNETH SHERMAN and E. G. SCHANER, U.S.
Bureau of Commercial Fisheries Biological
Laboratory, Boothbay Harbor, Maine.
Seasonal occurrence, distribution, and breeding of
the chaetognath Sagitta elegans in coastal waters
of Maine.
The chaetognath Sagitta elegans Verrill in the
coastal waters of Maine undergoes seasonal fluctuations in abundance, progressing from a spring low
to a summer high and declining in fall and winter.
These fluctuations are related to the breeding cycle.
One generation of S. elegans is produced annually.
Breeding extends from spring through fall: the peak
is in late spring. Maturing specimens dominate in
winter (>80% of the population), and immature
specimens in summer (75% of the population).
Death of mature individuals after breeding appears
to cause the spring population decline. Areal differences in abundance are influenced by the dominant non-tidal drift moving southwesterly along the
Maine coast. In winter, when the drift is poorly
developed, S. elegans is evenly distributed along the
coast. Numbers increase progressively westward in
summer and fall with the development of non-tidal
drift; they reach an annual peak in the western
194
AMERICAN SOCIETY OF ZOOLOGISTS
area (Cape Elizabeth, Maine, to Cape Ann, Massachusetts) during fall. Concentrations of S. elegans
along the coast are the result of local breeding.
There is no evidence of large-scale transport of S.
elegans to local areas from other breeding centers
in the Gulf o£ Maine.
42
PHILIP LEITNER and PAUL LICHT, Saint
Mary's College of California and University of
California, Berkeley.
Physiological responses to high environmental temperatures in three species of microchiropteran
bats.
Body temperature (TB), oxygen consumption, and
evaporative water loss were measured at ambient
temperatures (TA's) between 35° and 45°C in three
bats, Myotis yumanensis, Tadarida brasiliensis, and
Antrozotis pallidus.
At TA's up to 40°C, TB's rose gradually and stabilized 1°-2.5°C above TA without apparent distress.
When T 4 = 41.5°C, T B was kept below 42.5°C by
intermittent panting, salivating, and licking. A T B
of 43.5°C was lethal in about one hour and few bats
survived more than 15-20 min. with a T B of 44°C.
Therefore, at T 4 's of 42.5°C or higher, survival depended upon maintaining T B at or below T A by extensive evaporative cooling and minimizing metabolic heat production by remaining quiet. Under
these conditions, resting bats could dissipate up to
200 per cent of metabolic heat by evaporation. High
relative humidities definitely increased the stressful
effects of high TA's, chiefly because of greater activity and the consequent rise in metabolic heat
production.
In the three species studied, the physiological responses to high TA's in the laboratory correlate well
with behavioral responses observed in natural roosts
on hot summer days. The appearance of physiological stress at TA's near 41 °C is consistent with
the tendency in nature to avoid TA's above 41 °C.
The laboratory data confirm that the TA's between
45°-50°C that commonly occur in parts of their
natural roosts in summer represent potentially lethal temperatures. (Supported by N.S.F. Grants
GB-2885 and GB-4207.)
43
DERRY C. LAWRENCE and ADDISON L. LAWRENCE, University of Houston.
Effects of temperature upon the mechanisms for active transport of monosaccharides in the intestine
of a chiton, Cryptochiton stelleri.
Using everted sacs, segments of proximal anterior
intestine and proximal posterior intestine were incubated for two or three consecutive 2i^ or 4 hour
periods. Initial concentrations of monosaccharides
were 1 /iM/ml of 3-0-methyl-D-glucose or D-galactose, with serosal and mucosal solutions being identical. The temperature progressions followed in
these serial experiments were 10°C to 15°C to 20°C,
5°C to 15GC to 25°C to 15°C. Final concentrations
were determined by radioisotopic techniques.
Results for 3-0-methyl-D-glucose in the anterior
intestine show that no serosal/mucosal (S/M) ratios significantly greater than 1.0 were developed at
5°C. Increments in temperature of 5°C resulted in
a steady increase in S/M ratio, through 20°C (ratios
were in the neighborhood of 3 to 1 at 20° C). At
25°C, however, the first thirty minutes saw very
rapid formation of S/M gradients, but after this
time no further increase was observed. Moreover,
after 4 hours at 25°C, return to 15°C showed the
gut's transport capacity to have been destroyed.
D-galactose in the posterior intestine did not give
S/M ratios significantly greated than one at 50°C.
However, 5°C temperature increments caused S/M
ratios to increase in an almost perfectly linear fashion (25°C ratios as high as 5 to 1 for 4 hours were
observed). Unlike the results with the anterior intestine, transport capacity was not significantly affected by 4 hours at 25°C.
The effect of temperature upon absorption of 30-methyl-D-glucose and D-galactose across the chiton gut indicate an active transport process. (Supported by NIH Fellowship 2-F2-GM-8812-02 and
Grant GB-3256 from NSF.)
44
F. J. VERNBERG and W. B. VERNBERG, Duke
University Marine Laboratory.
Thermal acclimation and cytochrome c oxidase activity in tissues of temperate and tropical zone
fiddler crabs (genus Uca).
Thermal acclimation has been demonstrated at
different levels of biological organization. Previous
work at both the whole animal and tissue level has
shown that there are metabolic differences between
temperate and tropical zone fiddler crabs. However, no data in relation to thermal acclimation at
the enzyme level are available on these crabs. Since
the respiration of most aerobic organisms is linked
to a cytochrome system, cytochrome c oxidase activity was investigated in the supraoesophageal ganglia (SEG) and muscle tissue from warm- and coldacclimated Uca pugilator from the Beaufort, N. C.
area, and with warm- and cold-acclimated U. rapax
from Florida and Puerto Rico. Enzyme assays were
made at 15° and 25°C. There were marked differences in enzyme activity in tissues from these populations. At 15°C enzyme activity in muscle tissue of
the tropical zone crabs was much lower than in the
temperate zone crabs and no acclimation was noted.
Enzyme activity of muscle tissue from the temperate zone crabs was significantly enhanced following
cold-acclimation. Cold-acclimation did not markedly influence enzyme activity in the SEG from
temperate zone crabs. In the tropical zone crabs,
however, the enzyme activity i.i Lhe SEG was decreased following cold-acclimation. (Supported by
NSF Grant GB 4600.)
45
S. JAYARAMAN and H. S. DUCOFF, University of
Illinois.
The response of McGill Black and of wild-type
flour beetles (Tribolium confusum) to temperature stress and to anoxia.
As part of a program to evaluate the influence
of residual radiation injury and of age on physiological performance, we are examining the response
of adult flour beetles to various types of stress. This
195
ABSTRACTS
report concerns the effects of exposure of normal
young adults to lethally high temperatures and to
anoxia. Of particular interest are the general sequence of events, and the much greater sensitivity
of the inbred McGill Black strain for all parameters
scored. Wild-type beetles tolerate 44° for at least
24 hours, but succumb within a few minutes to a
48° exposure. At approximately 46°, the beetles exhibit markedly increased physical activity for the
first 1 or 2 hours, and then become comatose; if
removed after 2-4 hours, most beetles appear to
recover after a few hours, but nevertheless die during the following 10 to 14 days. The same sequence
is seen in the McGill Blacks, but for temperatures
at least 2 degrees lower. When beetles are placed
in a nitrogen atmosphere, all physical activity stops
within one minute; nevertheless, even after 12
hours of anoxia, the beetles eventually recover.
"Knock-down" time is shorter, and recovery time
is much longer, in the inbred black strain. (Supported by Grant GM-10208, U.S.P.H.S.)
46
EUGENE V. NELSON and J. H. CAMIN, University of Kansas.
Cuticular critical temperature of the rabbit tick,
Haemaphysalus leporispalustris.
In studying the evaporative water loss from feeding stages of the rabbit tick, Haemaphysalus leporispalustris, we have found a critical temperature
lower than the surface temperature of the host
(3I°-33°C). This is surprising because earlier critical temperature determinations for other ixodid
ticks have generally been a few degrees above the
host temperature. Since earlier workers have primarily studied engorged ticks, we also measured
these stages and found them to be much higher
(40°-45°C). On the basis of these results and other
recent findings it now seems likely that the feeding
stages of at least several ixodid ticks have critical
temperatures below the surface temperature of the
warm-blooded hosts. This means that evaporative
water loss is relatively high while the tick is on the
host, which may be important in stimulating attachment and in concentrating the blood meal.
These results may help to explain the relatively
low critical temperatures and the lack of interspecific variation thus far reported for the Ixodidae.
(Supported in part by Grant AI-02487 from U.S.
P.H.S.)
47
JAMES L. OSCHMAN and BETTY J. WALL, University of Cambridge, England, and Western Reserve University, Cleveland, Ohio.
Rectal pad structure in the cockroach, Periplaneta
americana.
Interest in mechanisms of ionic and osmotic regulation has led us to a study of the organization of
the cockroach rectum. Primary attention has been
given to the rectal pads, structures thought to function in water and ion reabsorption. Recta from
normal and dehydrated cockroaches were fixed in
glutaraldehyde followed by osmium, embedded in
Araldite, and sectioned for light and electron microscope examination. Each rectum has six pads sepa-
rated by thin rectal epithelial cells. Both pad and
epithelial cells possess a cuticular intima on the
luminal (apical) side. Rectal epithelial cells possess
an extensive subcuticular space containing a fibrillar material. This space is larger in dehydrated
animals. Between the pad cells and the haemocoel
lies a layer of connective tissue containing neurosecretory axons and trachea, a connective tissue
space of variable size, and a muscle layer. Some
neurosecretory axons appear to terminate in the
connective tissue layer. The apical membrane of
the columnar rectal pad cells is extensively infolded
to form leaflets. On the cytoplasmic side of these
leaflets is a layer of repeating subunits similar to
that found in rectal papillae of Calliphora by
Gupta and Berridge (J. Cell Biol. 29:376, 1966).
In contrast to Calliphora, the leaflets in the cockroach rectum are closely associated with elongated
mitochondria. Lateral membranes of pad cells are
also infolded and associated with mitochondria, but
the infolds are not stacked as in Calliphora. Intercellular spaces have been found between adjacent
pad cells, but no connection has yet been observed
between these spaces and the connective tissue
space. (Supported by USPHS Fellowship 1-F2-GM24, 015-01 to B.J.W.)
48
LOU ANNE ROBERTS, University or California,
Los Angeles.
Evaporative water loss in the desert lizard Uta
stansburiana. (Introduced by Malcolm S. Gordon)
Evaporative water loss was measured as weight
loss of lizards held in tubular chambers under a
constant air flow of 400 cc/min. Measurements were
made at four temperatures (15, 20, 30 and 35°C)
and at low, medium and high humidities. The data
were analyzed by quadratic least squares multiple
regression (method of Wood, Biometrics 21:775,
1965). There was no significant change with size or
surface area within tbe small size range considered
(1 to 5 gms). Three variables which were found to
account for most of the regression effect provide
the equation:
mg H.O/hr = 0.032 T + 0.037 SD + 0.014 T
(male) — 0.347
where T is temperature (°C) and SD is saturation
deficit (mmHg). Coefficient of determination =
0.72. Standard error = 0.29.
49
S. RAGHUPATHIRAMIREDDY and JAMES W.
CAMPBELL, Rice University.
Enzymes of arginine metabolism in insects. Arginase.
Of 12 guanidino compounds tested, including 7guanidinobutyrate, only L-arginine and L-canavanine were hydrolyzed by cetyltrimethylammonium
bromide extracts of Platysamia gloveri fat body: Larginine was converted stoichoimetrically to urea
and ornithine by such extracts. This hydrolytic activity showed an optimum around pH 10 and was
markedly dependent upon added Mn". The levels
of arginase activity were determined in the fat
196
AMERICAN SOCIETY OF ZOOLOGISTS
bodies o£ P. gloveri, Antherea polyphemus, and
Blaberus cranifera at various developmental stages.
There is an apparent increase in activity during development: in B. cranifera, 6th instar nymphs have
an average o£ 65 units (/xmoles arginine hydrolyzed
per hr at 25°, 85 mM substrate concen., and pH
9.5) per g tissue which increases to 103 units in the
adults; in P. gloveri, the activity increases from 105
units in diapausing pupae to 2000 units in the
adults; and, in A. polyphemus, from 34 units in the
pupae to 300 units in adults. In both P. gloveri
and A. polyphemus, there is little if any increase in
activity, in either units/g tissue or units/mg protein, during the first 15 days development. The
major increase occurs after this time. Whether this
apparent increase level of arginase activity is due
to new enzyme synthesis has not been determined.
There is no inhibitor of the adult enzyme in early
developmental stages as determined by mixing experiments.
No ornithine transcarbamylase activity was detected in pupae of P. gloveri and A. polyphemus or
adult and nymphal stages of li. cranifera. They are
therefore incapable of de novo arginine biosynthesis which is consistent with the known nutritional requirement for arginine by many insects.
The fat body arginase could thus conceivably function in conjunction with ornithine transaminase to
convert arginine formed through protein catabolism to glutamate for utilization in general oxidative metabolism. (Supported by NSF GB-4524 and
USPHS 1-K3-GM-6780.)
50
WILLIAM D. SCHMID. University of Minnesota.
Natural variations in nitrogen excretion of amphibians from different habitats.
Analyses of ammonia and urea concentrations
were performed on samples of plasma and urine of
anuran amphibians from different habitats. The
technique of microdiffusion analysis proved to be
very adaptable to field conditions and provided the
means whereby specimens could be sampled soon
after capture. Aquatic species such as Rana seplentrionalis and R. clamilans had significantly
lower levels of plasma urea than semiaquatic species such as R. pipiens and R. sylvatica, which in
turn had lower levels than more terrestrial species
such as liufo hemiophrys and B. americanus. There
was also a greater tendency for urea to be partitioned into the urine of more aquatic species. Ammonia content of urine was higher in the aquatic
forms, but varied greatly within species depending
upon the immediate state of hydration of the specimen. High levels of urine ammonia were used as
the basis of critical evaluation of the ecology' of B.
hemiophrys. The adaptive values of natural variations in patterns of nitrogen excretion were discussed relative to interspecific differences in ecology
and water balance. (Supported in part by grants in
aid of research from the Society of Sigma XI and
from the Graduate School of the University of Minnesota.)
51
PRENTISS G. COX, Western Reserve University.
The role of peripheral nerve fibers in tail regeneration in the lizard, Anolis carolinensis. (Introduced by Sidney Simpson, Jr.)
Singer and Kamrin (Jour. Exp. Zool., 138611,
1955) demonstrated that lizard regeneration would
not occur in the absence of the spinal cord. Simpson (J. Morph., 114:425, 1964) further showed that
it was the ependymal portion of the spinal cord
which supported regeneration and that it could
induce tail formation. However, the role of peripheral nerve fibers in lizard tail regeneration has not
been previously investigated. In the work to be reported it has been found that removal of the spinal
cord by cauterization reduced the peripheral nerve
number by only 25-35% and yet complete failure
to regenerate occurred except for approximately
four segments of muscle. Removal of the three
pair of dorsal root ganglia immediately proximal
to the level of autotomy and the accompanying disruption of the ventral motor nerves reduced the
peripheral nerve supply by 80-100% and yet normal regeneration occurred. This seems to indicate
that even in the absence of peripheral nerves the
ependyma is capable of supporting normal regeneration but that in the absence of the ependyma near
normal numbers of peripheral nerves are incapable
of supporting regeneration. The ependyma as it
grows out in the regeneration process is accompanied by several hundred descending spinal nerve
fibeis. This work together with the demonstration
by Simpson that transplanted ependymal pieces
(i.e., pieces without nerve fibers) could induce tail
formation would tend to indicate that the lizard
tail is capable of normal regeneration in the complete absence of nerve fibers either peripheral or
central if ependyma is present. (Supported by
Grant GM-12,653 from the U.S.P.H.S.)
52
STEPHEN D. SMITH, University of Kentucky.
Galvanic induction of partial limb regeneration in
Rana pipiens. (Introduced by Ira D. Fowler)
Forelimbs of small adult frogs (Rana pipiens)
were amputated and implanted with silver-platinum bimetallic rods insulated except at the tips.
The silver end was at the wound surface in some
cases. In others, the platinum end was distally
placed. Other animals received separate implants
of short pieces of bare silver and platinum wire,
with the silver piece placed near the wound surface. Controls consisted of animals with either no
implant or implants of partially insulated silver or
platinum wire.
Only one of the control animals (one with a silver implant) initiated regeneration. Regeneration
was minimal with the separate implants (25%),
more frequent with the platinum end of a bimetallic couple at the wound surface (70%), and most
frequent (83.4%) with the silver end o£ a bimetallic couple at the wound surface. In three cases in
•the last named series, lobulated cartilages reminiscent of wrist and hand elements differentiated. The
results are ascribed to direct stimulation of the
cells, electrophoretic transfer of information-bearing molecules, or stimulation of nerves. (Supported
by a grant from the University of Kentucky Re-
ABSTRACTS
search Foundation and by NSF Grant #B6-22O2-R.)
53
PATRICIA C. BARKER, University of Oregon.
Fine structure of blastoporal cells of Rana pipiens
gastrulae.
The blastoporal groove of the early gastrula of
Rana pipiens was studied in ultrathin specimens
fixed in gluteraldehyde and embedded in Epon or
Maraglas. Wedge and flask-shaped cells line the
groove at Stage 10 and are similar in appearance
to those described earlier for Hyla regilla (Baker,
J. Cell Biol., 24, 95-116, 1965). At earliest invagination, blunt, irregularly shaped microvilli cover the
cell surfaces. A dense matrix of granular material
and fine filaments underlies the distal cell surfaces
and projects into the microvilli. Proximal to this
dense layer, the cell neck contains small vesicles
and cytoplasmic particles, as well as large, granular
bodies of varying density. Some of these bodies have
no limiting membrane; others are partially membrane enclosed. Membrane bound organelles similar to the beta vesicles described by Perry and
Waddington (J. Embryol. exp. Morph., 15, 317330, 1966) are grouped together in the neck. Yolk
platelets, much smaller than those found in the
main body of the cell, are interspersed with the
granular bodies and fill the rest of the cell neck
proximal to the areas just described. Lateral cell
junctions in the apical region are characterized by
parallel membranes separated by a 100-200 A gap.
Just proximal to this zone are regions of desmosomal attachment.
As the groove deepens slightly, flask cells become
confined to the base of the groove. Bundles of microtubules and microfilaments are now evident
oriented parallel to the long axis of the neck.
Wedge-shaped cells along the sides of the groove
also contain filaments extending across the cell
parallel to the outer surface. Groups of filaments
run into the desmosome plaques.
As has been suggested earlier for Hyla regilla
(Baker, ibid., 1965), microfilaments and microtubules may furnish the motive force for certain of
the morphogenetic movements involved in amphibian gastrulation. (Supported by Fellowship
F2 HD-18,010-03 from the U.S.P.H.S.)
54
ELIZABETH J. H. HARTLINE, Brown University.
Electron microscopic observations of in vitro myogenesis. (Introduced by John R. Coleman)
Cell cultures derived from twelve-day chick embryo leg muscle were fixed and embedded for electron microscopy in such a way as to obtain both
longitudinal and cross sections of the cultures. In
the first two days of culture, when proliferation is
the primary activity, two main cell types are distinguished, fibroblast-like cells and spindle-shaped
cells. These differ in their fine structure. The former contain rough endoplasmic reticulum, and the
latter contain many free ribosome-like particles
(some in chains), a small amount of smooth endoplasmic reticulum, and sometimes filaments in their
cytoplasm. Production of filaments and fusion of
197
the spindle-shaped cells, which becomes prominent
in the second day of culture, appear not to be sequentially related. Multinucleated myotubes are
present by the end of the second day. Thin filaments appear in these before thick filaments are
seen. Under appropriate fixation conditions, the
first thick filamentous structures seen are microtubules. In older syncytia. these appear in great
numbers around developing myofibrils. Over a period of several days, the filamentous material increases in amount and becomes organized into myofibrils which develop striations. Dense material appears at intervals along the myofibril but does not
become organized into coherent Z bands until periodic elements of the sarcoplasmic reticulum appear.
Other cross-striations appear after the Z bands, until typical adult myofibrils are seen. (Supported by
Grants HD-OOO47 and HD-00019 from the U.S.
P.H.S.)
55
ROBERT H. WARREN and KEITH R. PORTER,
Harvard University.
Electron microscopic study of developing molting
muscle in Rhodnius prolixus.
The cyclic differentiation of the abdominal intersegmental molting muscles of Rhodnius prolixus
has been described at light microscope resolution
by Wigglesworth (Quart. J. Micros. Sci., 97:465,
1956). We have followed the same events with the
electron microscope, in the hope of elucidating the
mechanism of fibril formation.
The muscles of the fourth stage nymph are dedifferentiated and non-functional. No myofibrils
are present; the sarcoplasm contains free ribosomes,
some mitochondria and scattered bundles of longitudinally oriented 50-70 A filaments of unknown
nature. The envelopes of the axial nuclei are covered with ribosomes.
Muscle development is triggered by the insects'
ingestion of a blood meal. Within 10 hours, the
number of ribosomes on the nuclear envelopes decreases, and the number of polysomes in the sarcoplasm increases. The 50-70 A filament bundles disappear. At 15 hours of development, thick and
thin myofilaments, and microtubules are present in
groups along the peripheries of the fibers.
The definitive myofilament lattice forms slowly
over a period of 5-7 days. The aligning mechanism
appears to be the formation of bridges between
thick and thin myofilaments. Associations of sarcoplasmic reticulum with infoldings of the fiber surface membrane to form diads can be traced from
the initial stages of development.
A postulated function of microtubules in formation of the initial myofilament lattices has been
tested by colchicine treatment of developing fibers
in intact insects. Preliminary observations indicate
that microtubule destruction does not alter initial
number or grouping of myofilaments. (Suppported
by USPHS Training Grant 2G 707-06.)
56
RICHARD MORSE, Western Reserve University.
The synthesis of ribosomal and transfer nbonucleic
198
AMERICAN SOCIETY OF ZOOLOGISTS
acid in the epidermis of an insect. (Introduced
by M. Locke)
During the larval life of an insect the synthetic
activities of the epidermis are correlated with the
alternating behavior of feeding and molting. While
the animal feeds in the intermolt period, the epidermis deposits endocuticle. Preparatory to ecdysis,
enzymes secreted by the epidermis digest the old
cuticle and the digestive products are returned to
the hemolymph nutrient pool. At the same time,
a new and larger cuticle is being made to replace
the old cuticle. This physiological picture suggested that there might be a large variety of proteins synthesized in a closely coordinated sequence
at molting. During the intermolt period, there may
be a steady synthesis of a smaller number of different proteins. In Calpodes ethlius (Lepidoptera) it
was found that each of these different phases of
activity began with a period of increased RNA synthesis (Lock et al., Science 149:437, 1965). One peak
of RNA synthesis occurs just before molting. The
other peak occurs at the beginning of the intermolt
period. The functional significance of these two
peaks can be better understood if it is known what
kinds of RNA are synthesized at the different stages
during the molt cycle. It has been found that ribosomal and transfer RNA are synthesized during
both peaks and during the intermolt period of
minimal total RNA synthesis. The relative proportions of ribosomal and transfer RNA which are
synthesized at these different stages will be discussed. (Supported by Grant CM 09960 from
U.S.P.H.S.)
57
L. J. PIERRO, J. E. COGH1LL and R. M. WOLKOWSKI, University of Connecticut.
Effects of deoxyguanosine on axial development in
the chick embryo.
Previous studies in this laboratory have indicated
that deoxyguanosine may produce disturbances in
growth and development of the trunk region including axial mesoderm segmented prior to treatment. Formation of axial structures from the nodal
tissue and segmentation of axial mesoderm appear
to proceed normally. Viewed at the organismic level, treatment at 3 days of incubation may result in
rumplessness; at 2 days, in a thoracic constriction.
Continuing studies of the regional morphological
response have yet to provide a satisfactory explanation of the pattern described, but observations
made at several levels of analysis argue against suggestions either of differential distribution of deoxyguanosine or limitation of its biochemical action to cells which have attained particular stages
in their differentiation. First, although deoxyguanosine treatment of 2 day chick embryos does not result in the rumpless syndrome, it does increase the
probability of such abnormalities when administered in conjunction with either actinomycin D or
puromycin. Second, although cell death does not
occur in the nodal region as a consequence of treatment with deoxyguanosine alone, nor with puromycin, extensive cell death is found in this region
after treatment with both deoxyguanosine and
puromycin. Third, studies with chick embryo explants show that incorporation of H*-thymidine
into DN'A over a 24 hour period is reduced at all
levels of the embryonic axis. Reduced synthesis of
DNA is presumably a secondary manifestation of
the biochemical action of deoxyguanosine. The precise biochemical action of deoxyguanosine in the
early chick embryo remains to be clarified. (Supported by Grant GB 1940 of the N.S.F.)
58
ROCHELLE M. WOLKOWSKI and LOUIS J.
PIERRO, University of Connecticut.
Effects of actinomycin D on axial development in
the chick embryo.
Previous studies in this laboratory have indicated
that segmentation of the axial mesoderm and formation of axial structures from nodal tissue are
disrupted in chick embryos treated with actinomycin D. Various evidence suggests that development
of the head and trunk regions including axial mesoderm segmented prior to treatment proceeds normally.
Since actinomycin D inhibits RNA synthesis in a
variety of systems, it is assumed that effects on axial
development are mediated through interference
with RNA synthesis. Supporting evidence from
studies involving explanted chick embryos (11-13
somites)
will be presented. Thus, incorporation of
Hs-uridine into RNA is reduced by 50% during a
20 hour culture period in presence of 0.75 ^g actinomycin D; formation of somites is reduced 80%.
Progressively smaller dosages permit more somites
to form and increased incorporation. Incorporation
of appropriate precursors into protein and DNA are
also reduced, presumably as secondary effects.
Further experiments demonstrate reduced incorporation of Hs-uridine into RNA in the head and
anterior trunk region during one hour exposure to
actinomycin D. Incorporation in these areas moves
towards control levels during the second hour. Incorporation into the posterior trunk region (posterior to the 12th somite) falls below control levels
during two hour exposure and remains reduced
until at least 20 hours. Data for H'-thymidine incorporation into DNA also indicate only a temporary reduction in head and anterior trunk whereas
the reduction in the posterior trunk region continues
at least 20 hours. The significance of these observations for the regional morphological response to
actinomycin D continues under study. (Supported
in part by Fellowship 1-F1-GM-33, 242-01 from the
U.S.P.H.S. and CB 1940 from N.S.F.)
62
LOUIS LE GUELTE, Research Division, North
Carolina Department of Mental Health.
Orientation of a spider (Zygiella x-notata Cl.) in its
web. (Introduced by P. N. Witt) (Motion picture)
Zygiella x-notata builds an orb-web with a retreat usually located above the web (Web A). But
the animal can be made to build a web from a retreat below (Web B). On both of these webs, the
spider, after catching prey, goes back directly to its
retreat.
If a Web A is turned around 180°, the spider goes
to the prey without hesitation, then tries to go back
199
ABSTRACTS
to its retreat. During 1-10 minutes, it is obviously
disoriented and wanders repeatedly in the original
direction of the retreat. This spider is in a conflict between going in the direction it came from
or going where it usually finds its retreat. Analysis of data shows that the latter factor is predominant but can be decreased by keeping the spider in
the reversed position or by multiplying the numbers of trials. Then the spider will go back where
it came from. Other experiments show the relative
unimportance of the dragline and of the asymmetric structure of the orb in the orientation of the
spider in its web. (Supported in part by a grant
from N.S.F.)
63
JIM W. DOLE, San Fernando Valley State College.
Homing and orientation in Rana
pipiens.
In a field study of the behavior of displaced Rana
pipiens, using a trailing device to follow movements, it was found that adults displaced relatively
short distances (to about 1 km) generally oriented
correctly in the homeward direction and usually
homed successfully. Animals displaced greater distances showed no evidence of homeward orientation. Blinded frogs displaced up to 800 m oriented
and homed as successfully as normal frogs, from
both upwind and downwind of home. Anosmic
animals appeared to be disoriented. (Supported by
Grant GB-3476 from NSF.)
orientation to occur, although the prerequisites
are more circumscribed. At increased separation
distances all of the cues are weaker and orientation
ceases. (Supported in part by the Honolulu Biological Laboratory of the U.S. Bureau of Commercial Fisheries, and by NSF GB 4791.)
65
LESTER TALKINGTON, The University of Wisconsin.
Bird navigation and geomagnetism.
New information about the navigating behavior
of homing pigeons, obtained by helicopter following, provides substantial evidence that the fundamental faculty for place-to-place navigation depends on some form of geophysical input which
involves the earth's magnetic field. A zigzag effect
found in homing flights seems to represent a succession of corrections in the flight course based on
sampling of data along path. Homing paths,
mapped to a high degree of accuracy, prove to be
local in character and analogous to initial orientation which is local in character, evidently depending in both cases on physical data which is local in
character. From aeromagnetic data for one portion
of the Ohio experimental area, a strong tendency
is shown for homing paths to skirt around or head
between magnetic anomalies. (Supported by Grant
GB-5259 from N.S.F.)
64
66
PHYLLIS H. CAHN, Yeshiva University and Long
Island University.
Some observations on the schooling of tunas. (Motion picture)
Euthynnus affinis (18 inches fork length), were
subjected to partial hydrodynamic and hydroacoustic (swimming sound) isolation while separated in
adjoining channels that permitted only visual contact. Two or three cement channels (size range,
2y2 t o 3 feet by 11 to 24 feet) were partitioned
with transparent plexiglas (rigid) or acetate (flexible). Of 22 fish tested, 18 survived the confinement.
The schooling orientation of two fish on opposite sides of a transparent partition occurred for
prolonged periods of time (several days) if no
major swimming speed adjustments were required
for the maintenance of visual attraction. That is,
both fish had to be of similar size, and also had to
be in a comparable physiological state. In the absence of partitions, fish that differed somewhat in
size schooled together, and were better able to adjust for greater swimming speed variations. The
adjustment was often accomplished by executing
figure-eight movements, but with partitions, there
was insufficient space for this to occur. With widened channels, fish-to-fish attraction diminished.
This diminution occurred despite the fact that the
fish probably could still see one another at the separation distances used (visual acuity studies of Nakamura, personal communication).
It is proposed that in tuna schooling visual cues
are augmented by hydrodynamic and hydroacoustic
stimuli. If the separation distances are not too
great, these stimuli are of sufficient intensity for
NED A. SMITH and C. L. RALPH, University of
Pittsburgh.
Some characteristics of heart accelerating substance
from the cockroach ventral nerve cord.
The work described here is a portion of a comparative study of neuro-endocrine heart accelerating substances of the cockroach. Ventral nerve cords,
consisting of all thoracic and abdominal ganglia,
were dissected from adult male Periplaneta americana (L.), washed, and placed in cold or boiling saline. The tissues were then homogenized in glass
tissue grinders. Homogenates were centrifuged in
various experiments at 1000 X g. 11,000 X g, and
25,000 X g- All fractions were subsequently tested
for their heart accelerating activity on in vitro
heart preparations (Ralph, J. Insect Physiol. 8:431,
1962). In all experiments almost all of the activity
was associated with the soluble fractions. In experiments where tissues were boiled, soluble fractions
initiated acceleration during the first 30 seconds of
the assay. Heart response to varying concentrations
of the crude 1000 X g supernatant was found to
be linear over a range of frequency increases from
0 to 100%, and sometimes as high as 200%. Ultracentrifugation of the 1000 X g supernatant on a 4
to 40% gradient of sucrose in saline indicated that
the activity is associated with light fractions, as
was the case for corpora cardiaca extracts (Smith
and Ralph, Am. Zool. 6:575, 1966). The activity of
heat stable substances of the nerve cord is dializable, but exhibits some properties different from
the heat stable accelerators from the corpora cardiaca. (Supported
Health Service Fellowdiaca.
(supported by
by Public
Put
ship 1-F1-GM 33, 738-01/
200
AMERICAN SOCIETY OF ZOOLOGISTS
67
TSUN-SUNG CHEUNG, University of Miami,
Florida.
Endocrine control of growth and reproduction in
the stone crab, Menippe mercenaria (Say).
Various destalking experiments on adult decapod
crustaceans in the past had shown that such operations would either accelerate molting in some or
ovarian growth in other species. Few attempts had
ever been made to explain their difference. According to Weitzman (Am. Zool. 4, 329, 1964) destalking Gecarcinus lateralis accelerated ovarian development in spring when such development is
dominant, but accelerated molting in the fall, when
molting is dominant. These results showed that destalking only accelerates events that characterize
the animal at the time the operation is performed.
Results on the stone crab indicated that in August
at the summit of the ovarian developing season,
destalking of intermolt crabs led to precocious
molting. In November, after the spawning season
when the majority of crabs molt, destalking of intermolt crabs again accelerated molting. With the
approach of: spring in February, destalking intermolt females led to precocious spawning. In all
these months, however, eyestalkless crabs surviving
sufficiently long 'beyond a molt tend to spawn instead of molting. Also, when destalking was performed at the postmolt period, spawning and not
molting took place. Thus, in this species the consequence of destalking depends also on the molting
stage in which the operation is performed. The experimental animals were housed in similar isolated
tanks provided with Tunning sea water, at temperature and salinity fluctuating as in nature. The
animals were fed daily.
68
ANN HEFFINGTON BUNT and EBERT A. ASHBY, University of Texas Southwestern Medical
School.
Ultrastructural study of the sinus gland in the
crayfish, Procambarxis clarkii.
The sinus gland in the eyeslalks of decapod crustaceans is composed of terminals of neurosecretory
cells lying in the X-organ and perhaps elsewhere
in the central nervous system. It serves as a storage
and release site for five or more tropic factors which
are secreted into the contained blood sinus.
Ultrastructural studies of the terminals and distal regions of the axons discloses five different types
of neurosecretory granules, unmixed within the
endings, and distinguishable either in dimensions,
shape, electron density or staining reaction, fit situ
stimulation of the sinus gland and subsequent
study of the fine structure indicates a lower incidence of certain granule types, changes in the elementary granules, and evidence on the mechanism
of release, when compared with contralateral controls. The secretion process includes fusion of the
granule membrane with the plasma membrane of
the axon terminal with probable fracture of the
fused zone. The granule core next appears extracellularly, within the basement membrane, as a region of electron dense material having a diffuse periphery and lacking a limiting membrane. Nor-
mally the secreted material may enter the hemocoel
after diffusing across the basement membrane.
The observed granule types may correspond to
the stored form of tropic hormones isolated from
other crustaceans. However, the different types of
granules observed could represent stages in the
maturation of a common granule form. The latter
alternative is considered least probable. (Supported
by Grant NB-06623-01 from U.S.P.H.S.)
69
JOHN L. MCCARTHY, LINDA WESSON and
ROBERT G. GAGUANO, Southern Methodist
University.
Action of cyanotrimethylandrosterone on adrenal
microsomal reaction in various species.
The synthetic androstane 2a-cyano-4,-t,l7a-lrimethyl-17fi -hydroxy-5-androsten-3-one (cyanotrimethylandrosterolone, cyano-ketone, CTA) was
shown to reduce the conversion of pregnenolone to
progesterone by rat adrenal homogenate in vitro
(Endocrinol. 79: 1123, 1965). Tn this study, the effect of CTA on the 15,000 X g supernatant of rat
adrenal homogenate was examined. The adrenal
fraction, containing microsomes, was capable of
converting pregnenolone to deoxycorticosterone
(DOC) in vitro. When this microsomal fraction was
incubated with the cyano-ketone (0.14-0.36 ^M),
either under oxygen or nitrogen the conversion of
pregnenolone to corticoid product was partially or
completely blocked. The data provide further evidence that the cyano-ketone inhibits adrenal 3/3hydroxysteroid dehydrogenase and acts as a noncompetitive inhibitor.
The 15,000 X g supernatant fractions of homogenates of bovine, guinea pig or rabbit adrenal glands
were incubated with CTA. Whereas 0.14 pM CTA
inhibited pregnenolone utilization by the rat adrenal preparation, the effective levels of cyanoketone in the bovine, guinea pig and rabbit adrenal
fraction ranged from 1.4 to 14 ^M. While a higher
concentration of cyano-ketone was required to depress the conversion of pregnenolone to progesterone by the 15,000 X g supernatant of homogenate
of bovine, guinea pig or rabbit adrenal tissue, the
agent did appear to inhibit the 3/S-hydroxysteroid
dehydrogenase activity. (Supported by USPHS
Grant AM-05744.)
70
K. V. PRAHLAD and L. E. DELANNEY, Wabash
College and Northern Illinois University.
Induced metamorphosis in the Mexican axlotl:
Rectification of a genetic disability by thyroid
hormone.
Failure of the pituitary-thyroid axis prevents
normal metamorphosis in the Mexican axolotl.
This genetic deficiency is corrected by a single administration of a small dose of tri-iodothyronine.
The following preliminary investigations were undertaken to establish this altered status in the pituitary-thyroid axis of the postmetamorphic axolotl.
(1) Comparative study of the pituitary and thyroid
glands from pre- and postmetamorphic forms. (2)
Effects of thyroidectomy on skin molting. (3) Sequential changes in premetamorphic skin grafted
201
ABSTRACTS
onto both intact and thyroidectomized postmetamorphic animals.
There were uncertain variations in the histology
of the pituitaries of both pre- and postmetainorphic
forms and the thyroid follicles of both were filled
with colloid. However, the lining epithelium was
cuboidal to columnar with prominent nuclei in the
postmetainorphic axolotis whereas the epithelium
was reduced toward the squamous type in the premetamorphic thyroid gland.
The first sign of stoppage of skin sloughing in
thyroidectonii/ed postmetainorphic axolotis was observed between day two and eight (postoperative
period) and complete inhibition of skin sloughing
by twenty to twenty-seven days.
Premetamorphic skin when grafted onto a postmetamorphic intact animal snowed metamorphic
changes and became histologically similar to the
host skin in about thirty-five days. However, the
grafts on thyroidectomi/ed animals did not show
any changes except in the dermal fibrous connective tissue network.
Hypophysectomy of postmetamorphosed axolotis
resulted in cessation of molting, indicating that the
administered hormone had elevated the pituitarythyioid axis to a functional level. (Supported by
Grant GM 05619 from the USPHS.)
ROBERT A. COOPER, VIRGINIA L. JENTOFT,
and SEFTON R. WELL1NGS, University of Oregon.
A dish for hanging-drop organ culture, with particular reference to endocrine tissues.
Hanging-drops have been used for a variety of
purposes, including organ culture (Biggers, |. D.,
et al., J. Physiol. (Lond.) 131:497, 1956). This method has not, however, been widely used for a variety
of reasons, including poor drop stability and poor
fluid:tissue ratios. Because of certain advantages inherent in the method, a dish was designed to overcome these problems. Glass rings (0.5 cm in internal diameter; 0.45 cm in height) annealed to the
internal surface of standard petri dish covers produced wells with an average volume of 0.180 cc.
Using 1-2 mm tissue specimens, an approximate
fluid:tissue ratio
of 60:1 was produced. Siliconization (Siliclad8) of the wells increased drop stability, facilitating inversion and righting of the dish.
Using this dish, with medium changes at 48 hour
intervals, 1-2 mm fragments of testis, endomelrium,
mammary gland, and vagina, and whole ovaries and
adrenals, from 14 day old Balb/cCrgl mice were
cultured. Cultures were maintained at 36°C for
7 days in chemically defined medium 199.
containing 5 ^g/ml of amorphous insulin (20 IU/
mg, Eli Lilly Co.), in a humidified 95% O.:5% CO,,
atmosphere. Cell survival and structural preservation was comparable to standard methods. Use of
the above dish for organ cultures should offer several advantages, one of the most important of which
would be isolation of individual fragments or organs, thus allowing for in vitro studies of endocrine
hormone biosynthesis and metabolism not possible
with other techniques. (Supported by U.S.P.H.S.
Grant HD-00104 and Medical Research Foundation
of Oregon Grant 639.)
72
ALVARO CUADROS and ROBERT A. COOPER,
University of Oregon.
Ultrastructure of spontaneous vaginal keratinization
in organ culture (Balb/cCrgl mice).
Hanging-drop organ cultures (see abstract #71)
of vaginal mucosa from 8.5 week old mice, oophorectomized 10 days previously, were incubated for 4
days in chemically defined medium 199 at 36°C
in a humidified 95% O2:5% CO™ atmosphere. By light microscopy, the 3 cell layered cuboidal epithelium of the castrates increased in
thickness in culture and by the fourth day was
fully keratinized. Digested PAS stains demonstrated mucin secreting surface cells on days 1, 2,
and 3, while stains for sulfhydryl and disulfide
groups (Barnett and Seligman, J. Nat. Cancer Inst.
13:215, 1952; 14:769, 1953) became progressively
more positive from day 2 to day 4. As compared
with the oophorectomized controls, ultrastructural
studies at 24, 48, 72, and 96 hours in culture, using
OsO, fixation and araldite embedding, demonstrated increased prominence of nucleoli, desmosomes, and ribosomes, and progressive increases in
cytoplasmic fibrils, composed of 70A filaments. Unlike in vivo keratinization, keratohyaline granules
were absent in vitro; and, although comparable
changes in cell membranes and desmosomes (modified and composite desmosomes) occurred, the complete aggregation of filaments seen in the stratum
corneum in vivo did not take place in vitro. Thus,
keratinization in vitro appeared to be incomplete,
and the processes of in vivo and spontaneous in
vitro keratinization were shown to be not completely comparable. (Supported by U.S.P.H.S.
Grants CA TI 5081-06 and HD-00104 and by Medical Research Foundation of Oregon Grant 639.')
73
JERRY P. FRIEND and WENDELL \V. LEAVITT, University of Cincinnati.
Preparation of rat pituitary monolayers as a means
of studying the effects of hypothalamic extracts
on pituitary cells.
Monolayers of rat pituitary cells were prepared
using a modification of Moscona's technique for
embryonic tissue.
Adult male CD rats were lightly etherized, decapitated, and the adenohypophysis carefully extirpated under aseptic conditions. The glands were
serrated, transferred to Ca-Mg free Hanks' solution (CMFH), then added to a beaker containing
0.025% Pronase in CMFH, and allowed to digest
for 3.5 hr at room temperature without agitation.
Tissue fragments were then rinsed in 3 changes of
CMFH for 1 hr. Care was taken throughout to
prevent premature disruption of the tissue. Cells
were dispersed in a 40 ml centrifuge tube containing 5 ml CMFH by agitating with an Adams C)clomixer for 30-60 sec. The cell yield varied from 1
to 2 X 10° cells/pituitary. Cells were added to
CMRL 1066 or medium 199 (supplemented with
10% whole egg ultrafiltrate and 20% calf serum)
at 0.5 to 1 X 10° cells/ml medium. Cultures were
incubated in 5% CO2-95% air at 37°C. Confluent
monolayers were obtained after 7 or 8 days.
202
AMERICAN SOCIETY OF ZOOLOGISTS
Autoradiographic techniques have been employed to study effects on nucleic acid and protein
synthesis after addition of hypothalamic or cerebral cortex extracts. Preliminary results indicate
that RNA synthesis is increased in cultures incubated with hypothalamic extract but not in control cultures incubated with cerebral cortex extract.
Studies are in progress on the effects of hypothalamic extract on protein synthesis and hormone release in cultures with or without RNA and protein
inhibitors. (Supported by USPHS Grant HD01568
and Am. Cancer Soc. Grant IN79.)
74
FRANK E. FRIEDL, University of South Florida.
The partition of externally detectable nitrogenous
material from the aquatic pulmonate snail Lymnaea itagnalis jugularis (Say).
As part of a more extensive study on excretion
in Lymnaea, the distribution of nitrogen (assayed
as ammonia) was determined in material accumulating in the fluid environment (a dilute salt solution) of snails held at 25°C for 18-30 hours. In investigations on a total of 29 snails (average whole
weight, 2.06 g) pooled in five experimental groups
it was found that ammonia constituted about 52%
(39-71%) and urea about 28% (25-32%) of the total nitrogen detectable. As yet unidentified material represented the remainder of the nitrogen
(4-29%), and of this fraction, 75% (50-100%) appeared to be non-volatile material, with the difference possibly representing volatile material in addition to the above-mentioned ammonia.
Nitrogen-containing material accumulated at an
average rate of 3.4 micrograms nitrogen per gram
whole weight per hour (range 1.2-5.4); with the
rates of appearance of ammonia and urea corresponding in orders of magnitude with values found
in other similar studies performed with and without added antibiotics. The partition of nitrogenous material also is generally comparable with the
percent values reported by Spitzer (Zool. Jahrb.
57:457, 1937) for a twenty-four hour period at summer room temperature. (Ammonia-N 42.3%, UreaN 13.8%, Uric Acid-N 5.2%, Undetermined-N
38.5%).
It is hoped that closer scrutiny of the unidentified material will assist in further characterizing
the pattern of nitrogen catabolism in this snail.
(Assisted by NSF Grant GB 3158.)
75
ANNE FUNKHOUSER and SUSAN FOSTER, University of the Pacific.
Oxygen consumption during tadpole metamorphosis.
Eggs- of Hyla regilla were collected from ponds
near the Pacific Marine Station, Dillon Beach, California, and raised in the laboratory. Animals were
maintained in pond water at room temperature
(about 22°C) and fed boiled lettuce ad libitum until they reached the non-feeding period. Oxygen
consumption of groups of tadpoles (0.5 to 0.9 total
animal weight) was measured with Warburg respirometers using 15-ml well-less vessels containing
10% KOH in the side arm as a carbon dioxide absorbant.
Metamorphosis, from hatching to froglet, was
completed in about 30 days. Individual tadpole
weight increased until the end of prometamorphosis, then decreased as feeding stopped. The percentage of dry weight increased significantly during
metamorphic climax. Oxygen consumption of individual tadpoles (wet weight) is directly proportional to change in body weight. Oxygen consumption per gram wet or dry weight decreased from the
time of hatching, with the most pronounced decrease occurring just before feeding began, remained relatively constant during prometamorphosis, then increased during metamorphic climax.
Oxygen consumption of adults was lower than that
of juveniles. The transitory increase in oxygen uptake observed during metamorphic climax is
thought to result from the increased thyroid activity of the immediately preceding stages.
76
GEORGE F. CROZ1ER, University of Southern
Mississippi.
Chemical basis for a morphological color change in
a marine teleost.
The young of the California Sheephead, Pimelometopon pulchrum, were found to be quite different in coloration from the adults of the species.
Extraction of the integumentary carotenoids and
separation of fractions through column and thinlayer chromatography indicated that there was a
significant difference in the balance of the major
components.
Astaxanthin (3,3' dihydroxy, 4,4'diketo /3-carotene) was the predominant fraction of the adult
skin but was much less than tunaxanthin (tentatively—3,3' dihydroxy e-carotene) in the young. The
presence of a preponderance of tunaxanthin in the
skin of the latter made them conspicuously golden,
particularly when compared to the red shade of
the adults.
Controlled feeding experiments indicated that
the fish are capable of converting the tunaxanthin
to astaxanthin through zeaxanthin (3,3' dihydroxy
/3-carotene) as an intermediate. This report represents the first instance of chromophore conversion
by fishes, and is further, a heretofore unknown in
vivo reaction. (Supported by N.S.F. GB-2312.)
77
MORRIS H. BASLOW, University of Hawaii.
The function of N-acetyl-L-histidine in the fish
eye.
Acetate "C and L-histidine "C, when injected
into the anterior chamber of the eye of the carp,
Cyprinus carpio, produced labelled N-acetyl-L-histidine in the lens after as short a time as 30 minutes. However, glucose "C and pyruvate 14C do not
label N-acetyl-L-histidine even after 48 hours. Labelled histidine in the lens reaches a peak after two
hours and disappears over a period of 15 hours
during which time increased label in N-acetyl-Lhistidine is found. Maximum labelling of N-acetylL-histidine appears between 15-20 hours from either
acetate or histidine and labelled N-acetyl-L-histi-
ABSTRACTS
dine is slowly lost from the lens over a period of
days. When incubated in vitro in a lens support
medium, pre-labelled N-acetyl-L-histidine leaks out
as a function of time, with specific activity of "C
label varying inversely with time, while other free
amino acids are retained by the lens.
L-histidine "C has been found to be actively
transported into the lens from the surrounding
fluid and concentrated against a gradient, whereas
N-acetyl-L-histidine "C is not. Since no acylase activity is found in the lens, although it is high in
ocular fluid, a cycling phenomenon is indicated by
which ocular fluid histidine is regenerated from
lens N-acetyl-L-histidine. It appears therefore, that
the function of N-acetyl-L-histidine in the lens is
to serve as a carrier molecule for the passage of Lhistidine from tissue to the extracellular fluid. As
far as is known, this is the first description of an
energy driven amino acid recycling phenomenon
between a tissue and extracellular fluid involving a
change in molecular form, and the first demonstration of a lens metabolic sequence dependent on
an ocular fluid enzyme. These observations indicate that there is a potential histidine pump mechanism operating at the lens surface although the exact nature of the transfer phenomenon involved is
as yet obscure. (Supported by Grant GB-3607 from
N.S.F.)
78
PHILLIP SHEELER, San Fernando Valley State
College.
Stromal protein components of mature and immature red blood cells.
Immature red blood cells (reticulocytes) from
anemic rabbits (New Zealand white) and turtles
(Pseudemys elegans) incorporated plasma iron into
hemoglobin when incubated in vitro for 60 min. at
37°C, whereas mature red cells (erythrocytes) did
not. A small and relatively constant percent of the
iron removed from the plasma was recovered in the
hemoglobin-free stroma (primarily membranes) of
lysed cells. Chaser experiments employing plasma
labeled with iron-59 showed that iron was temporarily retained in the stroma but was subsequently
utilized in heme synthesis. In order to characterize
the protein components of red cell membranes, cells
labeled with iron-59 were lysed and the stroma
solubilized in urea, mercaptoethanol, sodium desoxycholate and sodium lauryl sulfate. The dissolved stroma was then fractionated on DEAEcellulose columns and the elutcd protein fractions
identified by their absorption at 260 m^; fractions
containing iron-59 were identified using a scintillation counter. Eight protein fractions were demonstrated in rabbit reticulocyte stroma and 11 in turtles; the iron-59 was associated with several of these
fractions. Starch-gel electrophoretic patterns of
stromal proteins from mature and immature red
blood cells were compared in order to identify any
major changes in the number and/or concentration
of components which accompany cell differentiation.
(Supported in part by N. I. H. Postdoctoral Fellowship 1-F2-AM-25, 672-01.)
79
DOLCLAS K. ANDERSON and ROBERT
L.
203
HAZELWOOD, University of Houston.
Characterization of avian cerebrospinal fluid. (Introduced by A. L. Lawrence)
Ceiebrospinal fluid (CSF) was obtained from
anesthetized female White Leghorn chickens by
aspiration of the cisterna magna with a tuberculin
syringe and 25 gauge needle. The needle with stylet was inserted, the latter then removed, and syringe subsequently attached; a maximum of 0.35 cc
blood-free CSF was obtained. Immediately thereafter, a blood sample was obtained by cardiac puncture. CSF and plasma were analyzed for total reducing substances, total protein, and amino nitrogen (colorimetrically), sodium and potassium (EppendorC flame photometer). Chickens were grouped
as 6-10 weeks old, 11-18 weeks old, and 1-2 years
old. Results indicate that CSF glucose tends to decrease with increasing age (136 ± 5.0 to 119 ± 10.3
mg%), although plasma glucose was essentially unaltered. Plasma/CSF glucose ratios of 1.93, 2.32,
and 2.06 were calculated for the three age groups,
respectively. Similarly CSF total protein levels decreased from 174 ± 14.6 mg% in 6-10 week old
birds to 123 ± 7.5 mg% in 1-2 year old birds. Respective plasma/CSF ratios were 25.4 and 37.6. CSF
sodium levels did not fluctuate among age groups,
approximating 154 mE/1; plasma/CSF ratios varied
between 0.97 and 1.03. Plasma potassium levels exceeded those in CSF (4.15 ± 0.14 vs 3.74 ± 0.11
mE/1); potassium ratios of 1.11 and 1.33 were observed for 11-18 week old and 1-2 year old groups,
respectively. Data obtained indicate that in age
groups studied a maturing regulation of CNS environment is achieved for glucose and total protein.
(Supported by GB-2752 from NSF.)
80
HONG S. MIN, Georgia Institute of Technology.
Entrance of carbohydrates into human cancer cells
grown in tissue culture (KB cell line).
The entrance of carbohydrates into the cells of
KB cancer cell line has been studied, using labeled
monosaccharides. At 10 mil external concentration
the rate of increase of intracellular concentration
decreases continuously and the maximum intracellular concentration never exceeds the extracellular
concentration. It appears that the kinetics of penetration resemble the Michaelis-Menton Law rather
than the Fick's kinetics. On the other hand, the
pattern of entry of sugars between 1.0 and 5.0 mSI
external concentrations does not resemble the predicted pattern of entrance by diffusion, but instead
the rate of entrance is constant up to [C],/[C]O of
1.80.
The data indicate that the transport system has
a high affinity for sugars and the saturation of the
system occurs very rapidly at 10 mM external concentration. The enzyme-like kinetics observed in
these studies suggest that a carrier mechanism may
be involved in the transport o£ sugars into cancer
cells (KB). The large value for V m « (18 mM/min)
and the rapid attainment of the steady state intracellular concentration by the KB cells suggests that
the cells have a large demand for metabolizable
sugars.
It appears that the mode of carbohydrate en-
204
AMERICAN SOCIETY OF ZOOLOGISTS
trance into KB cells is dependent upon the external
concentration of sugars and a carrier system involved. (Supported by Grants GM-12235 from the
U.S.P.H.S. and N'sG-657 from NASA.)
81
MICHAEL N. GOODMAN and ROBERT L. HAZEL WOOD, University of Houston.
Myocardial actomyosin and subcellular phosphorous levels in the alloxan-diabetic rat. (Introduced
by M. S. Harry)
Studies were undertaken to determine whether
cardiac actomyosin and subcellular phosphorous
fractions are altered in adult male Long-Evans rats
which were fasted or injected with alloxan. The
duration of fasting and of the diabetic state was investigated over a period of five days (fasting), and
over 3, 7, 14, 30, 60, 90, 120, and 210 days (alloxandiabetes). Actomyosin was determined by viscosimetric methods and as per cent of total protein.
The following cell fractions were isolated for phosphorous determinations: total homogenate, supernatant (cytoplasm), nuclei-myofibrillar, mitochondrial, and microsomal.
Fasting: Blood glucose decreased immediately
subsequently to rise toward normal levels by the
third day of tasting; heart/body weight ratios followed an inverse pattern simultaneously. Actomyosin, as measured by ATPase activity, decreased from
87.7 to 73.1 units after two days of fasting and
then increased toward normal by the fifth day of
starvation. Mitochondrial phosphorous levels decreased from a control level of 0.133 to 0.115 mg/
gm wet weight after 120 hours of starvation.
Alloxan-Diabetes: Fed blood glucoses ranged
from 396-550 mg%; heart/body weight ratios increased and then returned toward normal by the
end of the observation period. Actomyosin decreased from 87.7 to 69.7 units after the first week
of alloxan-diabetes, then increased toward normal
levels. Alterations in mitochondrial and microsomal phosphorous were noticed, with slight alterations in supernatant and nuclear-myofibrillar
phosphorous.
Characterization of the actomyosin, as a criterion
of purity, was determined by disc electrophoresis.
(Supported by Grant #210-5-65-66 from the Houston Heart Association.)
82
JOHN J. B. ANDERSON and LARRY D. BAKER,
University of Illinois.
Distribution of radiostrontium in the skeleton o£
the young pig with age.
Two piglets (Yorkshire-Hampshire cross) were
injected two days after birth with 1 ^tCi of strontium-85 each. The piglets were killed at 24 and 50
days after injection, at which time whole-body radioactivity measurements were made. The entire
skeleton was dissected out and individual bones
were ashed at 600° C for 24 hours. The ash was dissolved in 1:1 HC1 and diluted to volume (50, 100
or 200 ml), from which duplicate 15-ml aliquots
were removed for gamma spectrometric counting
and duplicate 2-ml samples for gravimetric determination of total calcium by oxalate precipitation.
The percent of the injected dose of strontium-85
per gram of calcium was calculated from the data
of the percent of the injected dose and grams of
stable Ca in each bone. The sum of the percent of
the injected dose for all the bones was slightly less
than the percent retained in the body as measured
by whole body counting, leaving only about 1 to
2% of the injected dose in blood and soft tissues.
The findings were: (1) that bones with high turnover rates, such as carpals, tarsals and some vertebrae, lose a large portion of their radioactivity between post-injection days 24 and 50; and conversely
(2) that bones with low turnover rates, such as phalanges and radii, retain a large portion of their
radioactivity between post-injection days 24 and
50. The relationship between individual bones and
the entire skeleton for the percent of injected dose
per gram Ca may be shown graphically. (Supported
by Grant AT (11-1)1339 from the U.S.A.E.C.)
HELEN GHIRADELLA, JAMES CRONSHAW,
and JAMES CASE, University of California, Santa Barbara.
Fine structure of the aesthetasc hairs of Coenobila
clypeatus.
Light microscope studies have revealed that the
aesthetascs of Coenobila clypeatus contain the dendrites of many bipolar neurons of which the cell
bodies are grouped in spindle-shaped masses beneath the bases of the hairs. The axons of each
group run centripetally, joining with those of other
groups to form the antennular nerve.
The hairs are partially recumbant so that each
has an exposed and an unexposed side. The electron microscope has revealed that the detailed morphology of the hairs reflects this asymmetry; the
cuticle on the exposed side is thinner and more
tenuous than on the opposite side. Furthermore,
all the dendritic elements are concentrated on the
exposed side, lining the thinner cuticle, while the
rest of the lumen contains a close packed mass of
other cellular elements which may be Schwann or
supporting cells.
The dendrites show well-developed rootlets and
basal bodies and terminate in cilia with the usual
9 -}- 2 structure. Before entering the hairs, the
cilia divide dichotomously until there are a number of branches, each containing one or more microtubules. These branches are then continuous
throughout the rest of the length of the aesthetasc.
Dye penetration studies have revealed that the
aesthetascs are extremely permeable and in this respect are similar to those of the marine hermit
crab, Pagurus. In view of the morphology of these
hairs and their permeability, it is probable that
they function as chemoreceptors. We hope that
study of the morphological differences between the
Pagurus and Coenobila aesthetascs may give some
insight into the different problems posed by chemoreception in aquatic and terrestrial environments. (Supported by University of California Facculty Grants 289 and 380 and Grant USPHSNB04372.)
87
LOIS G. KRUSCHWITZ, University of Hawaii.
205
ABSTRACTS
Aspects of the behavior and ecology of a reef
shrimp. (Introduced by E. S. Reese) (Motion picture)
Saron marmoratus, an aggregating, nocturnal
shrimp, occurs on the reef flats in Hawaii. Little is
known of the behavior and ecology of Indo-Pacific
shrimp. The habitat and temporal patterning of
activity, reproductive biology and larval development, the aggregations and social behavior, and
the specific uses of the appendages were studied.
At night, aggregations of a few to several hundred
animals graze on algae. At dawn, Saron retreats
into reef crevices, a prerequisite of its habitat. The
reef flats are bathed with ocean water, and salinity
and temperature variations are small. Saron shows
sexual dimorphism and breeds throughout the
year. The larvae metamorphose after about 30
days at 25-27°C in normal sea water. The aggregations and the social structure were studied under
laboratory conditions. Groups of males, females, and
both sexes were compared in a "standard environment"; males tended to distribute themselves more
evenly than females. In further experiments animals established territories in aquaria; then new
shrimp were added, and the resultant behavior recorded. The possibility of an "aggregating" pheromone was tested using a double Y maze and shrimp
discriminated water containing other shrimp. Appendages are used in specific ways; for example,
during locomotion there is a characteristic "sweepangle" of the walking legs. The chelate pereiopods
are used characteristically during feeding and
grooming. The grazing aggregations of Saron have
their ecological equivalent throughout the animal
kingdom, and the social structure of the aggregations appears convergent with that of grazing mammals. (Supported in part by Grant GB-3651 from
the NSF.)
DANIEL L. HOFFMAN, University of Washington.
Symbiosis between shrimp and anemones. (Introduced by E. N. Kozloff)
Observations have been made on the Hippolytid
shrimp, Lebbeus grandimanus, that appears to live
in close association with several species of anemones. This is the first such reported association
between temperate and sub-arctic species. Limbaugh et al. (1961) have seen similar associations
between the tropical cleaner shrimps, Periclimenes
pedersoni and P. yucatanicus, and the anemones,
Bartholomea annulata and Condylactis gigantea.
The shrimp were maintained in an aquarium
with nine different species of anemones and many
other species of shrimp.
Lebbeus grandimanus show a definite preference
for the anemones Taelia crassicornis, T. lofotensis
and Cribinopsis sp.
On introduction to the aquarium the shrimp approach the anemones and congregate in groups under the oral discs; usually attaching themselves to
the columns by their pereiopods. As a shrimp begins to move up the anemone, it prods the outer
tentacles with its third maxillipeds and first pereiopods. The tentacles of the anemone attach to the
appendages; the shrimp does not dart away but remains stationary. The tentacles soon relax and the
shrimp climbs higher onto the oral disc. The anemone reacts then relaxes until the shrimp is completely among the tentacles. It appears that once a
shrimp has mounted an anemone there is no response to any future Lebbeus that moves onto the
anemone. At times three shrimp have been seen
on a single Taelia crassicornis. However other species of shrimp evoke an aggressive response when
they contact an anemone.
Lebbeus appears to be feeding on egested matter
from the anemone and decaying tissue sloughing
off from the surface of the anemone.
The shrimp is strikingly marked with alternating
fluorescent bands of red, yellow and blue. It is
also probably gaining protection from predators
while in association with the anemones.
89
RICHARD V. BOVBJERG, University of Iowa.
Feeding and dispersal in lymnaeid snails.
Lymnaea stagnalis, Stagnicola reflexa, Stagnicola
exilis, Stagnicola elodes, while primarily herbivorous, feed selectively on carrion. Dispersal is not '&••
rected toward plant food but is strongly so toward' •
animal food. Chemoreception and homing is clearly manifested in weak concentrations of animal filtrates while non-existent in plant filtrates. Feeding
behavior is elicited in far lower concentrations of
animal filtrates. Sites of chemoreception include
all of the head and foot region but in decreasing
sensitivity laterally, and posteriorly; the tentacles
appear to act as homing devices. Responses to food
stimuli include rhythmic buccal movements, lateral
bending, and clasping-flexing of the foot. Again,
animal tissue stimuli elicit the stronger responses.
The picture emerges of snails feeding continuously
on abundant plant material without directed movements, but actively homing on the occasional carrion in the habitat. Evidence is presented to support the notion that some animal food is needed
for normal growth and reproduction.
90
HERMAN T. SPIETH, University of California,
Davis.
Lek behavior of endemic Hawaiian Drosophila.
Field studies conducted in various parts of the
world show that the typical activities of feeding,
mating, and ovipositing by species of the genus
Drosophila occur diurnally in the early morning
and the late afternoon. Individuals of all ages and
both sexes gather for relatively short periods of
time on discrete, usually small, masses of fermenting materials such as decomposing fruits and fungi,
slime fluxes, etc. At such sites the gregariously assembled individuals, usually consisting of a number of species, engage in feeding, ovipositing, courting, and mating. The females spend much time
feeding whereas the males spend a lesser time feeding. Males who are sexually mature devote most of
their time at such diurnal assemblages in courting
and mating.
The members of the rich endemic Hawaiian
Drosophila fauna differ in that both courtship and
mating are spatially and temporally separated from
feeding and ovipositing. On the feeding and/or
206
AMERICAN SOCIETY OF ZOOLOGISTS
ovipositional sites the males restrict their activity
merely to feeding, and individuals of both sexes
are cryptic in behavior and appearance at such
times. The sexually mature males do, however,
assemble on lek territories in the near vicinity of
feeding and ovipositing sites. Each male vigorously
defends his lek territory, at the same time advertising his presence to sexually receptive females.
Significantly the males of the various species possess striking sexually dimorphic characters. The
evolution of this lek behavior, as well as certain
other unique behavioral characteristics of the Hawaiian species, can best be explained as an adaptive response • to predator pressure, exerted probably by the native Hawaiian honeycreepers.
91
ING. JEAN MATHIEU, Tnstituto Tecnologico y
de Estudios Superiores de Monterrey, Mexico.
Mating behavior of five species of lucanidae (Coleoptera: Insecta) (Motion picture)
Observations mainly on sexual behavior were
made in both field and laboratory on five species
representing four genera all occurring in Central
Illinois.
In males of Lucanus Scopoli ( = Pseudolucanus)
and Dorcus MacLeay, which have relatively short
mandibles, there was a direct use of these appendages in subduing and maintaining the female in
position during mating. In the species studied there
was a relatively high level of sexual activity translated in a high rate of copulation, in some instances up to 30 matings per hour.
Males of Platycerus Geoffroy and Ceruchus MacLeay having more dimorphic mandibles do not use
these appendages to hold tightly the female, but
rub intermittently a pair of legs against the female's
elytra and have much longer matings.
The presence of a female-stimulus was determinant in the amount of fighting among males. When
males were exposed to the presence of a female
and the latter was taken out, a gradual extinction
of the fighting drive was recorded, appearing in
many instances as homosexual interactions.
Lucanus placidus, a nocturnal species, revealed
a male population density of over ten times that
of the females. Groups of fighting males were
commonly found, often a female had dug into the
sandy soil leaving the fighting males on the surface. In all cases sex recognition occurred by contact of palps and antennae probably mainly
through close range olfactory cues.
92
ANITA ZORZOLI, Vassar College.
Gl uconeogenesis in mouse kidney cortex. Glucose
production and enzyme activities in newborn
and perinatal animals.
Gl uconeogenesis in kidneys of 0 (newborn), 7,
14, 21 and 28 day-old C57B1/6 mice was studied in
2 ways: (1) by measuring in vitro production of
glucose by cortex slices; (2) by determining the activities of 6 enzymes involved in gluconeogenic
pathways.
At birth glucose production from either oxaloacetate or succinate precursor wTas well below the
adult level (equal to 100%). Synthesis from oxaloacetate rose to 120% at 7 days of age, reached 160%
at 14 days and then declined to about adult value
at 28 days. With succinate precursor, glucose production rose gradually to slightly above 100% by
28 days.
The specific activities of glucose-6-phosphatase,
fructose 1,6-diphosphatase and fumarase were low
at birth, being less than 40% of adult values. By
28 days each enzyme had reached or exceeded
100%. Lactic dehydrogenase and triosephosphate
isomerase were near 100% at birth, declined slightly at 7 days and then rose to 100%. The developmental pattern for phosphoenolpyruvate carboxykinase was quite different from the patterns of the
other enzymes. From 64% of adult value at 0 days,
specific activity rose to 134% at 7 days and to 310%
at 14 days. At 28 days activity was down to the
7 day level. The glucose production and enzyme
activity data presented here can be interpreted as
supporting the hypothesis that phosphoenolpyruvate carboxykinase has a rate-limiting role in renal
gl uconeogenesis. (Supported by Grant HD-00494-08
from the U.S.P.H.S.)
93
FRANK SETO, University of Oklahoma.
Analysis of liver and spleen enlargements during
the graft-versus-host (GvH) reaction in chick
embryos.
The degree of hepatomegaly was found to be directly proportional to splenomegaly among 14-day
recipient embryos grafted with cells from allogeneic
donors (Seto, Growth 30257, 1966), but it was not
resolved whether the parallel liver response was
mainly a GvH-elicited proliferation as in the
spleen, or involved a nonspecific enlargement secondarily associated with splenomegaly. White Leghorn recipient embryos were treated in the following ways: (1) one group was given cyclophosphamide a day before donor cell injection, (2) another
group received methotrexate and blood, and (3) a
third group was grafted on the chorioallantoic
membrane (CAM) with spleen or liver pieces from
donor embryos previously injected with adult Game
fowl blood. Cyclophosphamide treatment reduced
hepatomegaly to a lesser extent than splenomegaly,
whereas methotrexate treatment while effectively
suppressing splenomegaly simultaneously accentuated hepatomegaly. The hepatomegaly in embryos
with CAM spleen grafts, when compared with the
relative liver to spleen enlargements in control recipients, was more closely correlated with the combined enlargements of CAM graft and host spleens
than with the host splenomegaly alone. Liver grafts
produced negligible GvH responses. The differences in the growth kinetics of spleen and liver of
chemically treated recipients and of CAM-grafted
recipients indicate that the liver enlargement involves both GvH proliferation and splenomegalydependent nonspecific responses. (Aided by Grant
E-409 from the American Cancer Society.)
94
E. PETER VOLPE and BRYAN M. GEBHARDT,
Tulane University.
ABSTRACTS
Mixed stem blood cell populations in the bone
marrow of ex-parabiotic leopard frogs.
The authors demonstrated previously (Science
154:1197-1198, 1966) that postmetamorphic diploid
and triploid frogs that had earlier been joined in
parabiotic union from embryonic life until metamorphosis were each chimeric with respect to their
blood cells. Chromosome preparations of cultured
peripheral blood (drawn from the femoral artery)
revealed the presence of both diploid and triploid
leucocytes in each ex-parabiont. The existence of
circulating leucocytes of two genetically distinct
kinds in ex-parabionts that have been apart for as
long as six months or more signifies that blood
cell precursors were interchanged when the exparabionts shared a common circulation in embryonic life. The critical question that has lingered is
whether or not the donor-type primordial blood
cells actually home in the hematopoietic tissues of
the host.
A definitive answer was sought by chromosomal
analysis of bone marrow cells of the ex-parabiotic
frogs. Cytological preparations of marrow cells from
the femur and tibiofibula were made following the
technique of Seto (Cytologia 30:437-446, 1965).
Both diploid (2n = 26) and triploid (3n = 39)
metaphase plates have been found in the marrow
cell preparations of ex-parabiotic frogs. Evidently,
the peripheral blood cell chimerism previously encountered is a permanent condition. That is to
say, donor-type stem blood cells do actually settle
in the blood-forming tissues of the host and perpetuate their kind indefinitely. Postmetamorphic
ex-parabiotic frogs are highly tolerant of skin
grafts from their former partners. This specific
mutual acceptance of skin homogafts is attributable
to the permanent blood cell chimerism. (Supported
by Grant GM-11782 from the PHS and Grant IN24-1 from the American Cancer Society.)
95
GORDON M. RAMM, University of Maryland.
The vitelline circulation of chick embryos subjected to prolonged low centrifugal stress.
The chick embryos subjected to low levels of
centrifugal stress for prolonged periods of time respond with an array of structural and physiological
anomalous conditions depending upon the magnitude and duration of the stress and the time of
application. One of the initial responses is the production of an abberant vitelline circulation.
Series of fertile eggs were subjected to centrifugal
stress between approximately two and eight "G"s
from their initial incubation to six days. The vitelline circulation showed a retardation of total development and aberration of the major elements of
the system. Gross observable variations from a
standard norm never exceeded ten percent in controls but all experimental series showed statistically
significant numbers of abberant patterns in the
order of fifty percent of two "G" animals by the
sixth day and reached one hundred percent in
series at higher "G" levels after the same period.
At higher levels death usually occurred earlier in
development and the alfects were greater.
Embryos were removed from centrifugal stress
after incubation for 24 to 120 hours and allowed
207
to continue to develop to six days. Animals surviving the treatment showed significantly greater ability to recover if treatment was less than 72 hours
and did not exceed 4 "G"s. Some of the recovering
animals showed a vascular pattern approaching normal.
In part the vitelline failure can be correlated
with the activity of the cells in the angioblastemal
region and the accumulation of blood cells in the
sinus terminalis.
96
GERALD J. MIZEJEWSKI and GORDON M.
RAMM, University of Maryland.
Phagocytic activity in the thyroid glands of the
chick and mouse embryo.
Mouse embryos, ages fifteen to twenty days, were
injected with colloidal carbon by cardiac puncture.
The embryos were maintained in mammalian
Ringer's at 37.5°C for fifteen and thirty minute intervals. Chick embryos, ages three to twenty days,
received intravenous injections of colloidal carbon
via the chorio-allantoic circulation and were incubated for similar intervals. The neck region of
both embryos was prepared histologically and examined to determine the phagocytic capabilities of
the embryonic endothelium of the thyroid.
In both the avian and mammalian embryos,
blood spaces appeared in the developing thyroid
prior to and during follicle formation. In the
chick embryo, the thyroid primordia from the sixth
to eighth days possessed blood spaces which exhibited phagocytic capabilities. By mid-incubation,
the blood spaces had developed into a sinusoidal
circulation capable of greater particle uptake. Conversion of the sinusoids to a capillary circulation
occurred during the third quarter of incubation.
By the seventeenth day a capillary circulation had
been established in the thyroid accompanied by a
decrease in the carbon uptake. At the time of
hatching, particle ingestion in the thyroid endothelium had diminished.
In the mouse embryo, organ histogenesis occurred
mainly in the fourth quarter of gestation. A blood
space circulation, developed in the primordia from
the fifteenth to seventeenth days, displayed phagocytic abilities comparable to the pre-sinusoid stages
of the chick embryo. The sinusoidal condition was
never attained in the mouse thyroid and carbon
uptake diminished during the eighteenth and nineteenth days. During this time, capillary networks
became more abundant and replaced the blood
space circulation. At the time of birth, the phagocytic ability of the thyroid endothelium had been
reduced.
97
ALBERT L. ETHERIDGE, The University of Texas.
Mesonephric kidney determination in the newt,
Taricha torosa. (Introduced by A. G. Jacobson)
The development of the kidneys has often been
studied, yet little is known about the cause of determination of these organs. Many studies have implicated the pronephric duct as an important in-
208
AMERICAN SOCIETY OF ZOOLOGISTS
ductor of the amphibian mesonephric kidney. However, in the absence of the duct abnormal and nonfunctional tubules do sometimes form. Studies in
amphibians and other animals have also shown
that parts of the nervous system interact with the
mesonephrogenic mesenchyme to augment tubule
formation. It is obvious that most determination
of the kidney occurs prior to induction by the pronephric duct. This is similar to lens elicitation
where induction by the prospective retina is obviously important, but most lens determination has
occurred prior to retinal association. The actual
complete history of interactions between mesonephrogenic mesenchyme and other tissues in the
environment has never been worked out and an
evaluation of the state of determination at early
developmental stages is also missing.
My experiments are designed to measure the
state of determination of the mesonephrogenic
mesenchyme from earliest development and to
trace the histories of important interactions between this mesenchyme and other tissues in its environment. The importance of the tissue interactions occurring between the mesonephrogenic mesenchyme and the notochord, endoderm, and neural tissue as well as the pronephric duct are being
studied. Explants of many combinations of tissue
from neurula stages 14-18 develop tubules in a
high percentage of cases indicating an extensive
highly determined kidney field. Defect experiments
at these stages give similar results. During tailbud
stages 20-26 important interactions are demonstrable between the mesonephrogenic mesoderm
and tissues in its environment. It is during these
stages that the kidney field becomes limited and
dependent upon induction by the pronephric duct
for tubule formation in vivo. (Supported by Grant
HDOO725-06 from the U.S.P.H.S. to A. G. Jacobson.)
98
J. ROSS STEVENSON, Kent State University.
Lack of correlation of some growth and developmental processes in the crayfish.
Development of new setae, reabsorption of old
cuticle, growth of gastrolithSj and regeneration of
pereiopods were studied during the premolt period
in the crayfish Orconectes sanborni. It was found
that reabsorption of old cuticle and secretion of
gastroliths always began before the beginning of
formation of new setae. On the other hand, premolt growth of regenerating limb buds did not always begin before the beginning of formation of
new setae. This suggests that premolt limb growth
may be initiated by a mechanism independent of
the mechanism initiating these other processes.
Rates of reabsorption of old cuticle, of growth of
gastroliths, and of formation of new setae were
found to vary independently in different individuals. For example, the membranous layer of the old
cuticle may be fully reabsorbed in an individual in
which new setae have just begun to form, and part
of the membranous layer may still be present in
another individual in which new setae have been
completed. Either animal may have the greater
gastrolith/carapace length ratio. This suggests that
different sets of factors may control the rates of
these three processes. (Supported by Research
Grant GM 08363-05 from the U.S.P.H.S.)
99
WAYNE KUENZEL and C. W. HELMS, University of Georgia.
Obesity produced in birds by electrolytic lesions
placed in the ventromedial hypothalamus.
Many migratory birds deposit fat in preparation
for migratory flight in the spring. This deposition
can be artificially elicited in the laboratory by
proper manipulation of the photoperiod (D. Earner In A. Wolfson (ed.), Recent studies in avian
biology, Univ. Illinois Press, Urbana, 1955). In mammals, experimental obesity can be induced either by
destroying the ventromedial hypothalamus or by
electrically stimulating the lateral hypothalamic
nuclei (B. K. Anand and J. R. Brobeck, Proc. Soc.
Exptl. Biol. Med. 77, 323, 1951; J. R. Brobeck,
Physiol. Revs. 26, 541-559, 1946; J. M. Delgado and
B. K. Anand, Am. J. Physiol. 172, 162-168, 1963).
Lipid deposition under these conditions is quite
comparable to that found in birds during the premigratory period (J. R. King and D. S. Farner. In
H. Whipple (ed.), Ann. N. Y. Acad. Sci. 422-440,
1965).
We have conducted a series of lesion experiments
on a migratory bird, the White-throated Sparrow,
Zonotrichia albicollis. Upon destruction of the ventromedial region of the hypothalamus we have obtained obese birds during periods of the annual
cycle when photoperiod is nonstimulating. Initial
gains in weight, due primarily to fat deposition resulting from conspicuous hyperphagia, were statistically indistinguishable from weight gains in photostimulated controls prior to vernal migration (1.51.6 g/week). These results implicate the lesioned
region of the avian brain in the normal control of
lipid deposition. (Supported by Grant AM 06798
from U.S.P.H.S.)
100
ROBERT SMALLEY and KATHERINE SMALLEY, Kansas State Teachers College.
Adipose tissue development in the hamster.
In the hamster, brown fat develops in a matrix
of unilocular cells which closely resemble the "signet-ring" cell of white fat deposits. All sites which
contain brown fat in the adult hamster are occupied by unilocular cells at 4 days of age. The unilocular cells are subsequently displaced by small
cells, possibly arising from the reticuloendothelial
system. We refer to these small cells, which occur
in closely packed clusters, as immature brown fat
(IBF) cells. The replacement of unilocular by IBF
cells commences at about 4 days of age and is virtually complete by 12 days. The IBF cell increases
in size, but does not become multilocular until
after 12 days of age. The major lipid accumulation
in the IBF cell commences at about 16 days. After
this time the cells become markedly multilocular
and lapidly come to resemble the brown fat cell of
the mature hamster.
White fat in the hamster (inguinal deposit) does
not develop from a multilocular cell similar to that
of mature brown fat. The development of the adipose tissues in the hamster thus does not appear to
ABSTRACTS
follow the currently accepted patterns in other
forms. (Supported by Grant AM 09839 from the
U.S.P.H.S.)
101
RAYMOND F. BLOUNT and ISABEL H.
BLOUNT, University of Texas Medical Branch.
Previously undescribed bodies in the thin segment
of the loop of Henle.
With certain treatments the cells of the thin
segment of the loop of Henle in the mouse lose
their squamous character. Large vacuolated bodies
are present that reach a size several times the normal diameter of the tubule. Following short treatments such as pitressin injection, a body consists of
a vacuolated stained globule with frothy appearance. At this stage it appears to be within a cell.
With treatment of longer duration, such as high
salt or dehydration, a body is more dense and darkly staining although it remains vacuolated. At this
time the staining is similar to that of the basement
membrane. In later stages, particularly in old animals, although the vacuolation remains on the
luminal surface, the density of the globular mass is
increased. In such bodies spaces appear, filled with
granules staining red with Schiff's reagent. A suggestive observation is that there is often an association between elements of the capillary net and
the tubule at the exact position of a body. In some
cases, what appear to be blood elements are within
the globular body. Their location is in the outer
part of the inner zone of the medulla, but it has
not been determined which limb of the loop they
occupy. These bodies have never been observed in
young normal mice but in normal old animals they
are a common finding. These changes indicate that
the thin limb is involved actively in the mechanism
of urine concentration. (Supported by Grant HE
07494 from the U.S.P.H.S.)
102
K. V. THIRUVATHUKAL, Canisius College.
Fine structure of the adrenal glands of the freshwater turtle, Chrysemys picta.
The purpose of this investigation was to describe
the fine structure of the adrenal glands of the
freshwater turtle.
The structure of the adrenal glands was studied
with routine histological stains. The presence of
sudanophilic lipids and cholesterol was demonstrated by Sudan Black B and the Shultz test, respectively.
The adrenal glands are paired bodies, irregular
in outline and yellowish in color, extending along
the ventromesial surface of the kidneys in a retroperitoneal position. The glands are not completely
capsulated, but there is an intermixture of renal
and adrenal tissue on the dorsal surface of the
glands. The ventral surface of the glands is covered by pleuroperitoneum, which at the ventrolateral edge gives rise to the mesentery supporting
the gonads.
The stroma of the adrenal gland consists of a
connective tissue capsule, composed of collagenous
and reticular fibers. The capsule, invaginating into
the parenchyma, divides the gland into small
209
masses of tissue. The gland consists of an intermixture of cortical and medullary tissue. The cells
of the cortex range from irregular polyhedral to
columnar cells. These cells show positive reaction
with Sudan Black B and the Shultz test, and they
have a uniform distribution of lipids. The medulla
shows two kinds of cells—outer, which are light
brownish yellow and are located at the periphery,
and inner, which are dark brownish yellow and are
embedded in the side of the cortical cords or in the
intercortical sinuses. Both kinds of cells are arranged singly, in small clumps or in strands.
103
PAUL T. MEDICI and P. L. DEBIASE, St. John's
University.
Hemopoietic protection against lethal X-irradiation by splenectomy.
Our preliminary studies (Amer. Zoologist 6:360,
1966) had revealed that splenectomy afforded protection to rats exposed to lethal doses of whole
body X-irradiation.
In this report the hemopoietic regenerative capacities of intact and splenectomized animals have
been evaluated to demonstrate a possible mechanism underlying such protection. In addition,
spleen cell suspensions were introduced directly into
the femur of intact irradiated rats to assess their
hemopoietic protective potential.
Rehabilitation of the bone marrow was more evident in animals splenectomized 10 days prior to exposure to 600 R/wb than in intact rats. Furthermore, the peripheral blood of splenectomized rats
was characterized by large numbers of reticulocytes,
leukocytes, nucleated erythroid and blastoid cells.
The results may indicate the absence of the normal inhibitory effect of the spleen on the maturation and/or release of cells from the bone marrow.
Thus, the augmented hemopoietic activity following splenectomy enables the animal to survive the
critical period which follows exposure to lethal
doses of X-irradiation.
Increased survival rates were also apparent in intact rats exposed to 725 R/wb when spleen cells
were injected directly into the left femur within 24
hours post-irradiation. Quantitative marrow analyses of a series of rats subjected to 600 R/wb revealed higher total cellularity values in the spleen
cell injected group than in noninjected irradiated
controls. These findings are in accord with those
of other investigators and support the concept that
spleen cells may provide cellular precursors that are
capable of repopulating hemopoietic tissues.
104
DAVID E. WILSON, Central College.
The relationship between cytoplasmic regeneration
and plasmogamy in Actinosphaerium nucleofilum.
Plasmogamy (cytoplasmic fusion of separate animals) was studied under a variety of conditions.
The general requirement for this phenomena to
occur is that the membrane and/or the axopods be
"sticky" or partially solated. This condition was
simulated by 1) attempts to capture prey, 2) contact with an air-water interface, 3) ionic stimula-
210
AMERICAN SOCIETY OF ZOOLOGISTS
tion, 4) electrical stimulation and 5) low hydrostatic pressure.
More drastic treatment with ions, electricity or
pressure resulted in radical fragmentation of the
animals. If left undisturbed in a suitable medium,
these fragments regenerated or reorganized themselves as smaller animals in a manner previously
described as cytoplasmic regeneration (Greef, 1869;
Penard, 1904; Kuhl, 1951).
These cytoplasmically regenerated smaller animals or satellites also had a "sticky" surface and
tended to fuse and form larger individuals whenever they made contact with one another. Because
cytoplasmic regeneration and plasmogamy share
this "sticky" condition, they are considered by the
author to be related phenomena. The significance
of these relationships will be discussed. (Supported
by Grant 5TI-70 from the USPHS.)
105
ANSAR HUSAIN, School of Tropical Medicine,
Liverpool, England.
Preliminary serological studies on stage specificity
in Brugia malayi. (Introduced by Perry C. Holt)
Larvae of B. malayi obtained from mosquitoes
(Aedes togoi) and adults obtained from hamsters
were washed in saline and transferred at pin point
to clean saline solution. Serum from rabbits was
obtained after 3 weeks of three injections, approximately 100 larvae per injection, every 10 days.
The larvae and adults separately were well crushed
and extracted in phenolized saline—0.85% sodium
chloride extract plus 0.5% phenol. Antigens were
preparaed in three dilutions, 1:25; 1:50; and 1:100,
and tested against rabbit serum using agar-gel
diffusion technique.
When adult antigen was used, three bands
were noticed with 1:25 dilution, two with 1:50, and
none with 1:100. With 1:25, one band is wide,
close to the well; another outside, and a thinner
band is in the middle. With 1:50 the middle thinner band is missing. With larval antigen four bands
are formed with 1:25 dilution, two with 1:50, and
none with 1:100. The location and nature of the
four bands is approximately the same as the adult
antigen bands except the fourth band is thin, just
outside the wide band near the well. No attempt
was made to resolve these bands. Serum absorbed
with adult material removes all three adult bands
and three of the larval bands formed at 1:25 dilution, the larval outside coarse band remaining. Serum absorbed with larval material removes all four
larval bands and two adult bands, the outside
coarse band again remaining. These investigations
suggest that there are qualitative differences between antigens in adult and larval stages of B. malayi.
106
JERRY L. PUTXAM and LAWRENCE S. DILLON, Texas A&M University.
Heart structure and the ancestry of amphibians.
An extant dipnoan, Protopterus annectens and
an extant amphibian, Rana pipiens possess several
similarities in heart structure which suggest common descent. For example, in both animals, the
typical tetrapodian vessel, the posterior vena cava,
is present; the opening of the pulmonary vein relative to the sinus venosus is similar; the atrial cavity
is partitioned into right and left chambers; and
the conus arteriosus has a 180° spiral, is partitioned
internally and has pocket valves located principally
in the proximal end.
Since most of these similarities are related either
directly or indirectly to the pulmonary mechanism,
it might be suggested that some have arisen independently in each group represented, as a result of
the pulmonary processes. However, a study of the
heart of two air breathing actinopterygians, Lepisosteus osseus and Amia calva fails to support this
contention. Neither does the heart structure of
Polyplerus bichir support this suggestion although
its lung morphology closely parallels that of lungfish and amphibians. Thus, the development and
use of the pulmonary apparatus seem independent
of heart morphology.
Consequently, on the basis of heart structure investigated in the present study, only the lungfish
can reasonably be assumed to show evidence of
common descent with amphibians.
107
HANS E. KAISER, The George Washington University and National Aquarium, Washington,
D. C.
Studies about the walking mechanism of the horseshoe crab Limulus polyphemus L.
The normal walking mechanism of Limulus is
on one of the five pairs of legs. As these walking
legs are covered by the animal's shell the rhythm
of walking is unknown. Many scientists believe
that the last pair of legs, notable by its size and its
structural arrangement, plays an important function in the forward movement. This may be underlined by the fact that the last legs of this animal possess special spreading appendages which
prevent these legs from sinking into the sand or
mud. We had a number of small and large specimens in our aquaria whose movements we observed
and recorded by means of motion pictures. Sometimes it was possible to observe the spreading of
the appendages in the large specimens in the big
tanks beside the glass window. We also observed
the powerful action of the last pair of legs in forward action. The function of the other four pair
of legs was mainly in balance. In a special aquarium (plexiglass) moving pictures from below were
taken too. From views of the different angles we
determined that the power of movement is found
largely in the last pair of legs. (Appreciation for
the assistance of Mr. Peterson, Acting Director,
The National Aquarium to this project is expressed.)
108
HENRY PANG, University of Wyoming.
Animal behavior in the future of sociology.
Sociologists have paid little attention to the
study of animal behavior, nonhuman interaction is
not considered a proper area of study. Yet, in animal and human collectivities lie many similarities
ABSTRACTS
and differences which need to be examined more
closely. There are many standard fields of sociology
where animal behavior studies can be integrated
and employed by sociologists. And many sociological research findings can be used by natural and
biological scientists in a similar fashion.
Other social scientists such as psychologists and
anthropologists have made important contributions
to animal behavior. Some of the possible reasons
for the inactivity of sociologists is discussed. Inasmuch as the study of animals in a social system is
sociological too, sociology can and should play a
part in the frontier science of animal behavior.
Since animal behavior is at best interdisciplinary,
exchange of theoretical and methodological studies
among natural, biological, and social scientists is
badly needed.
109
JOHN H. HOLL1S, University of Kansas.
Operant analysis of stereotyped movements.
Monkeys, chimpanzees, dogs, and children raised
in socially and physically restricted environments
frequently show a high free-operant rate of stereotyped movements. These environments are generally void of the contingencies and consequences
that control adaptive behavior. This paper presents a method for direct measurements of stereotyped movements (e.g., rocking) and provides data
on acquisition and control (Variable-interval and
Fixed-ratio schedules) of rocking movements in institutionalized profoundly developmen tally retarded children. The results show that some subjects (rockers) respond to Variable-interval and
Fixed-ratio schedules of reinforcement in a manner
similar to other operants (e.g., bar-pressing or keypecking). Other subjects show a deficient operant
response, i.e., they are relatively insensitive to contingency and consequence changes. In one subject
who had a zero free-operant rate of rocking, we
were able to generate stable rates and control of
rocking movements using operant procedures. (Supported by Grant No. 00870 04 from NICHHD.)
110
JOHN D. SPOONER, Augusta College.
Variation in sound production and in life histories
of different populations of the fork-tailed katydid, Scudderia furcala, in the United States.
(Tape recordings)
Three forms of Scudderia furcala occur in the
United States; one each in the Southwest, the Northwest, and all of the East. The eastern and northwestern forms are morphologically more similar
than either is to the southwestern form. Yet, distribution is continuous only through the southwestern form.
The eastern form of this katydid is represented
by three seasonally distinct populations. Adults of
the single generation in the North occur between
the period of adulthood of two populations in the
southern and eastern coastal plain.
Sexual pairs are formed by males and females
moving toward sounds made by the opposite sex.
Males produce a short phrase which attracts females. Males also produce a lisp which triggers a
211
tick-like sound from females. Males move toward
the female tick. Individuals from several localities
have been tape recorded and audiospectrographic
analysis reveals striking differences in acoustical behavior of individuals from different populations.
Ill
RICHARD S. PETERSON and GEORGE A. BARTHOLOMEW, University of California. Santa
Cruz, and University of California, Los Angeles.
Airborne vocal communication in three species of
pinnipeds.
During field studies of northern fur seals (Callorhinus ursinus, California sea lions (Zalophus californianus), and northern elephant seals (Miroutiga angustirostris), we have paid particular
attention to patterns of vocal communication. In
this paper we describe the physical characteristics
and apparent social significance of the airborne vocalizations of each of these three species in their
natural environments. This acoustic behavior is
then discussed in relation to the ecology of the animals and current theories of animal communication.
We have been able to catalog nine distinct calls
in Callorhinus, six in Zalophus and seven in Afirounga. The differences in repertoire seem to relate primarily to a) the complexity and kind of information being exchanged; b) the amount of ambient noise; c) the availability of channels for nonacoustic communication: and d) the physical characteristics of the sounds that the vocal apparatus
permits. (Supported by USPHS Grant MH-1143001, the U. S. Fish and Wildlife Service, and Johns
Hopkins University).
112
RONALD C. SIMONS, RUTH A. BOBBITT, and
GORDON D. JENSEN, Washington Regional
Primate Research Center and University of
Washington.
An experimental study of mother M. ncmestrina's
responses to infant vocalizations.
There are now several studies which catalog the
vocal repertoires of non-human primates, include a
description of the setting in which the vocalizations
were heard, and suggest their possible communicative significance. This study is based on such a
catalog (that of R. Grimm, from this laboratory)
and is an experimental test of the response of
mother monkeys to two infant calls.
Two different vocalizations from each of two infants and a control vocalization from an adult female have been tape-recorded, and from these calls
a standardized stimulus tape has been prepared.
These calls have been played to mother monkeys
separated from their infants and isolated in a
sound-proof room, to adult female non-mother
monkeys and to adult male monkeys. We have
measured the responses of our subject animals in
two ways: we have counted the numbers of vocalisations emitted by the subjects and, using photo
cells, we have measured the amount of pacing occurring during the vocalization stimulus periods.
To minimize experimenter bias, stimuli were pre-
212
AMERICAN SOCIETY OF ZOOLOGISTS
sented and responses recorded by a specially constructed automated device..
In this situation mother monkeys, and only
mother monkeys, respond regularly and consistently
to the stimuli with increased pacing and calling.
The intensity of the mother monkey's response is
not merely a function of high arousal level and
therefore an unselective response to any stimulation because, as her base line activity decreases over
time after separation from her infant, her responsiveness to the specific vocal stimuli increases.
113
DANIEL E. WILLARXJ and WILLIAM F. MARTIN, University of Texas.
The vocalizations of four sympatric species of Texas terns.
The vocalizations of four species of tern (Hydroprogne caspia, Thalasseus maximus, T. sandvicensis, Sterna forsteri) were recorded in the field on
the gulf coast of Texas. Analysis by sound spectrograph and oscilloscope showed that the calls of
all four species are composed of the same three elements: a carrier frequency, a frequency modulation and an amplitude modulation. The calls used
for like behavioral situations are similar in structure: the carrier frequency is 3.2-3.8 KCS., frequency modulations lower the carrier frequency
from 1.6-1.3 KCS., and the amplitude envelope is 2
KCS. wide. Immature terns do not make frequency
and amplitude modulations.
114
SOPHIE JAKOWSKA and P. PAUL FAGUNDO,
27 West 96 Street, New York, N. Y.
Nutritional studies in newts, Notophthalmus viridescens. II. Low caloric intake of high protein
diets.
Diets of approximately 0.03 calories per 0.1 ml
dose were freshly prepared using: desiccated
trimmed beef liver (Difco Bacto Liver), a soybean
product (Purina Assay Protein RP-100, 94% or
more protein and 0.3% or less fat), and a salmonoid
diet (Halver) of 38 parts casein, 12 parts gelatin, 9
parts corn oil, 28 parts white dextrin, 9 parts a-cellulose mixture with vitamins, minerals, and 200
parts water. Twelve weekly intragastric 0.1 ml
doses were given to groups of 10 males, collected
during autumn in North Carolina, uniformly distributed by weight (2.7-3.5 g), preconditioned unfed
for 2 weeks and kept at 12-15°C with 12-hour artificial daylight cycle. Unfed control group received
0.1 ml distilled water. All newts survived and were
sacrificed 3 days after the last dosing. Body weight
change was calculated as per cent of initial weight,
and organ weight as per cent of terminal body
weight. Averages are reported.
Body weight losses occurred in all groups: Unfed,
16.0%; Halver, 25.0%; RP-100, 13.0%; Liver, 9.0%.
Initial liver weight was high in these autumn
newts: it was not significantly affected, though
slightly higher in fed groups: Unfed, 6.93%; Liver,
7.39%; Halver, 7.96% RP-100, 8.0%. The empty
gastrointestinal tract (esophagus through cloaca)
was heavier but shorter in fed newts, especially
with Halver diet, 7.25% vs. Unfed, 6.08%; Liver,
6.68%; RP-100, 6.69%. With Halver diet the fat
bodies were visibly larger and heavier, 1.71%, vs.
RP-100, 1.05%; Unfed, 1.15%; Liver, 1.18%. They
were absent in five newts receiving RP-100, in two
of unfed and Liver-fed groups, and in one on
Halver diet.
115
SOPHIE JAKOWSKA and P. PAUL FAGUNDO,
27 West 96 Street, New York, N. Y.
Nutritional studies in newts, Notophthalmus viridescens. III. Studies with sucrose and two therapeutic nutriments high in carbohydrate or fat
contents.
Diets of approximately 0.3 calories per 0.1 ml
dose (comparable to a dose of 25 mg Difco Bacto
Liver: Amer. Zool., 6, Abstr. 375, 1966), were freshly prepared from sucrose and from Sustagen; the
latter consists mainly of carbohydrate (66.5%) and
milk protein (23.5%) with low fat content (3.5%).
Lipomul, an emulsion with approximately 66%
corn oil, used as received, supplied approximately
0.6 calories per 0.1 ml dose. Twelve weekly intragastric 0.1 ml doses were given to groups of 10
males, collected during autumn in North Carolina,
uniformly distributed by weight (2.7-3.5 g), preconditioned unfed for 2 weeks and kept at 12-15°C
with 12-hour artificial daylight cycle. Unfed control group received 0.1 ml distilled water. All newts
survived and were sacrificed 3 days after the last
dosing. Body weight change was calculated as per
cent of initial weight, and organ weight as per cent
of terminal body weight. Averages are reported.
Body weight decreased by 16.0% in unfed newts,
and by 10.0% in newts fed sucrose and Lipomul,
although the latter Teceived more calories; Sustagen-fed group showed 3.0% gain over the initial
weight, and organ weight as per cent of terminal
body weight. Averages are reported.
Body weight decreased by 16.0% in unfed newts,
and by 10.0% in newts fed sucrose and Lipomul,
although the latter received more calories; Sustagen-fed group showed 3.0% gain over the initial
weight. Liver weight was not affected in sucrosefed newts (6.61%, vs. Unfed, 6.93%), but was significantly higher with Lipomul (8.67%) and Sustagen (10.86%). In Lipomul-fed newts the empty
gastrointestinal tract was heavier, and longer:
9.41%, vs. Sustagen, 7.01%; sucrose, 6.28%; Unfed,
6.08%. Fat bodies were visibly larger and heavier:
Lipomul, 3.02%; Sustagen, 1.82%; sucrose, 1.51%;
Unfed, 1.15%. Spleen weight was not affected. No
differences were noted in duplicated ninhydrinpositive patterns on paper in descending butanolacetic-water system from uniform hydrolyzates of
heparinized whole blood and liver of 2 newts per
group from these and from groups on 0.03 calorie
diets.
116
ALLAN J. STANLEY, L. G. GUMBRECK and
J. E. ALLISON, University of Oklahoma.
Hereditary absence of the entire reproductive system of the rat.
A genetic mutation which obliterates the reproductive system of rats was discovered in an inbred
213
ABSTRACTS
kindred of King X Holtzman hybrids in our colonies, May, 1966. This anomaly confirmed by laparotomy has now been found in 15 animals. External characters resemble those of females. A shallow patent "vaginal" orifice or a scar is present in
the "vaginal" position. Nipples are visible throughout the animal's life. Body size and weight are intermediate between those of males and females.
Karyotypes show them to be genetic males, an X
and a Y chromosome being present in all cases examined. On the basis of these findings and on previous work in our laboratories, on male pseudohermaphrodite rats of which we have produced
more than 500 animals, the authors propose the
following theory of genie control of sex differentiation in mammals: one pair of genes is responsible
for development of gonads, one for reproductive
tract. Normal animals would possess the dominant
genotype GGRR or G-R- or GR (G for gonads and
R for reproductive tract). Pseudohermaphrodites
would have the genotype G-rr or Gr. Animals with
only a reproductive tract would have the genotype,
gg R- or gR and the animal lacking both would
be the homozygous double recessive, ggrr or gr. Sex
linkage is probable; if proven, two separate loci on
the X chromosome would fit the theory. A single
dose of either dominant or recessive gene could
produce its respective trait in males. (Supported
by grants, NIH HD-01075-01 and Institutional
Grant BI165605.)
118
J. B. K1TZMILLER and R. D. KREUTZER, University of Illinois.
Salivary gland chromosomes of some Central American Anopheline mosquitoes.
Salivary gland chromosome preparations of
Anopheles punctimacula,
Anopheles
apicimacula,
Anopheles eiseni, Anopheles triannulatus,
Anopheles oswoldoi, Anopheles aquasalis and Chagasia
bagnathus have recently been made from material
collected in the Canal Zone and in the Republic of
Panama. All species show a salivary complement
similar to species previously studied, namely one
pair of short heterosomes and two pairs of metacentric autosomes. Of the three species studied
thus far in detail, two, punctimacula
and apicimacula, belong to the subgenus Anopheles, and
conform, in the banding patterns of the autosomes,
to the pattern typical of that subgenus. T h e third
species, aquasalis, is similar in pattern to albimanus; both these latter species belong to the subgenus Nyssorhynchus. No inversions are present in
the individuals of any of these three species. All
three possess distinctive X-chromosomes; identification of each species is possible using the banding pattern of the X. (Supported in part by Gran/
E-3486 from the U.S.P.H.S.)
119
117
SARA H. FRYE, P. O. Box 267, Irvine, Kentucky
40336.
LAURENCE G. GUMBRECK, ALLAN J. STANLEY and J. E. ALLISON, University of Oklahoma.
Cytogenetic analysis of a X-ray induced, doublemarker (yellow-achaete) mutant in Drosophila
melanogaster: T h e next step.
Multiple effects of a new allele probably of the
albino series in rats. (Introduced by J. F. Lhotka)
Cytogenetic analysis of a X-ray induced (2-kr)
closely linked yet structurally unrelated doublemarker mutant (Y/iJl+ y ac sc+ In49 BM1 $) of
A new allele which appears to be one of the albino series which extends white, thus limiting color
distribution of any basic color pattern, was discovered as a mutant of cream hood in a single female
of our King X Holtzman hybrid rats. This animal was born on 10-26-62 and an extensive progeny
has been derived from this single source. We have
designated the allele as c1 (c denoting the albino
series and the superscript 1 denoting limitation of
color.) A second effect of this gene is the production of sterility in all males that receive it. By 30
days of age tubular damage is well under way
functional sterility is apparent by day 60 or shortly
thereafter and by day 120 all elements of the germ
line have disappeared completely. Such animals,
destined to become sterile, can be identified at
birth by the color pigment pattern in the skin. All
females possessing the gene are fertile. A third
effect of this gene is the production of heterochromia in a small portion of the offspring. This trait
seems to be largely limited to the female sex, only
an occasional male possesses the trait. These heterochromic males also become sterile by the time
of puberty with few exceptions. Our suggested arrangement for the albino series of multiple alleles
would be C, c r , c1 and c. (Supported by grants NIH
HD-01075-01 and Institutional Grant B1165605.)
two specific genetic regions originating in mature
sperm (i.e., haploid germ cells containing highly
polymerized X-chromosomal-DNA coated with protein) of young Drosophila males (of genotype Y/
J]\* y* ac* sc+ In49 BM1) indicated that the yellowachaete mutant is neither associated with a gross
structural change such as an inversion or a translocation nor capable of being rigorously correlated
with the light-microscope analysis of the doublemarker mutant's polytene salivary glands.
Interpretations (among others) of the structure
of the yellow-achaete mutant are as follows:
1. T h e two mutants are the products of the position effects of two minute inversions, and represent
two dissimilar minute rearrangements. These mutants would be very unlikely to give crossing-over
with one another without either deficiency or duplication (Offerman, 1935).
2. Pilot reverse mutation experiments (sc*.Y/
4-kr
v
i j l * y ac sc+ In49 BM1 $ X Y / y f : = J ) indicated that the yellow mutant was non-revertable
(physical deletion or complete irreparable inactivation?) and that the achaete mutant was revertable
(position effect of minute inversion or partial reparable inactivation?). If one of the double-marker
mutants was a deficiency (non-revertable, non-sepa-
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AMERICAN SOCIETY OF ZOOLOGISTS
rable either via crossing-over or X-ray induced
breakage) and the other mutant was a position effect of a minute inversion (revertable, separable
via crossing-over and/or low-dose X-ray induced
breakage, then it would be difficult to demonstrate
with techniques now available the double mutant's
precise chromosomal structure in view of the extremely small (and equal) sue of these two specific
regions partially based on the forward and back
mutation data of American and Russian workers.
Additional analysis to establish structural and
functional equality among similar mutant phenotypes such as y ac or ac sc of independent origin is
needed. These problems are neither simple nor
solved. (Supported by the estate of the author's
deceased father and a grant from U.S.P.H.S.)
120
CLYDE MANWELL and C. M. ANN BAKER, The
Laboratory, Citadel Hill, Plymouth, England.
Polymorphism of turbot hemoglobin: a "hybrid"
hemoglobin molecule with three kinds of polypeptide chains.
Though fnost vertebrate hemoglobins have the
quarternary structure a2/32, heterozygotes for a hemoglobin polymorphism doA not
display "hybrid"
molecules of the form a./3 /3B in electrophoresis.
The paradox is emphasized by the occurrence of
such "hybrid" molecules in many enzyme polymorphisms (J. Mar. Biol. Assoc. U. K. 47: 145
(1967)). The hemoglobin situation has been explained by Guidotti and colleagues by symmetrical
dissociation combined with molecular separation.
Yet, a similar failure to find naturally mixed molecules in individuals heterozygous for immunoglobulin allotypes arises from cell specialization involving suppression of one of the two alleles.
Individual turbot (Scophthalamus maximus)
have any one of three hemoglobin electrophoretic
patterns: a single "slow" band, a single "fast"
band, or three hemoglobin bands. The "hybrid"
hemoglobin forms spontaneously from a mixture
of pure "slow",and "fast" and has been isolated
and shown to consist of two a and one each j3F and
'/3s chain. The gene frequency of "fast" hemoglobin
is 0.40 in both Plymouth and Aberdeen turbot,
suggesting "balanced polymorphism." This particular lysine-glutamate mutation forms a counterexample to universal application of the symmetric
dissociation theory and is concordant with other
data on the existence of "hybrid" hemoglobins in
some but not all interspecific hybrids (Comp. Biochem. Physiol. 10: 103 (1962)). There exist in proteins a few critical sites were mutation can alter the
quaternary structure (see also Science 152: 1393
(1966)).
121
J. K. SHERMAN, University of Arakansas.
Freeze-thaw induced latent injury as a phenomenon
in cryobiology.
Freeze-thaw survival or injury of cells and tissues
usually is assessed soon after thawing. Part of freezethaw induced injury appears latent (J. Dairy Sci.
42:94, 1959) and, therefore, escapes detection un-
less a sufficient post-thaw interval precedes evaluation.
Lettre ascites tumor cells were trozen slowly
(3.5°C/min.) in the vapor of solid CO3 (—78°C)
and thawed rapidly (33°C/min.). Following 0, 4
and 8 hours storage at 22°C, split samples of unfrozen and frozen-thawed cells were evaluated in
terms of rate of oxygen consumption ()„., mitotic activity, and plasma membrane integrity, by
differential respirometry, cell reproduction after
transplantation, and eosin staining. Two month
old white mice were used in four parallel experiments.
Data showed that: 1) In terms of Q02 reproduction, and eosin test, survival just after thawing (0
hr) was 86%, 67%, and 70%, respectively; 2) Extent of Qoa loss in 8 hr storage was much greater
in frozen-thawed than unfrozen cells (87% vs 39%),
with loss during the first 4 hrs accelerated almost
seven-fold (71% vs 9%); 3) Based upon mitotic activity, essentially all (93%) frozen-thawed cells perished after 8 hrs compared to 61% of the unfrozen
population, with loss in reproductive capacity in
the first 4 hrs doubled by freezing and thawing: 4)
Eosin test revealed definite but slight latent injury
during the second 4 hrs of storage, none during the
first 4 hrs, and an 8 hr loss of 46% for frozenthawed and 38% for unfrozen cells.
Results established latent injury as a cryobiological phenomenon. The complexity of survival evaluation was stressed especially in terms of variations
and relationships between criteria employed. The
significance of latent injury was discussed and need
for its evaluation with other cell types and in general pathology was suggested. (Supported by Grant
GM 06418 from U.S.P.H.S.)
121a
ALBERT F. EBLE, Trenton Junior College, Trenton, New Jersey.
The histochemistry of glycogen and some enzymes
concerned with its metabolism in the oyster,
Crassostrea virginica (Gmelin).
Glycogen is stored as aggregates of small granules in the cytoplasm of Leydig cells, being especially concentrated near the alimentary canal, digestive gland, mantle and gill. Hibernation results
in a decrease in total storage content, the amount
being roughly proportional to length of hibernation. Amylophosphorylase has a distribution similar to glycogen, with very high activities noted immediately adjacent to the developing gonad, the
digestive gland and the gut. High concentrations
of this enzyme are also present in the ventricle of
the heart and the adductor muscle. The shellsecreting region of the mantle also shows abundant
quantities of this enzyme. Amylo-l,4-» 1,6-transglucosidase (branching enzyme) has a distribution
similar to amylophosphorylase. Glycogen and the
above enzymes are presently being investigated
with respect to seasonal distribution in cells and
tissues including metabolic derangements due to
invasion of parasites. (Supported by Contract 1417-0003-137 from the U. S. Bureau of Commercial
Fisheries.)
ABSTRACTS
122
CENE GRUBITZ, III, University of Arkansas.
Agonistic behavior of male ground squirrels. (Motion picture)
Intraspecific agonistic behavior was observed and
filmed in six species of male ground squirrels (Citellns tridecemlineatus, C. mexicanus, C. spilosoma,
C. harrisi, C. leucurus, C. lateralis) in indoor and
outdoor enclosures and in the field. In initial encounters all species exhibited a stereotyped pattern
of agonistic approach and contact in which both
animals postured their bodies and held their
(luffed tails horizontally at a right angle to the
body. The two moved alongside one another in a
head to tail orientation with the tail of each being
in front of the face of the other. The two then began pushing at the flank of each other, and a revolving pattern resulted. Tail presentation toward
the opponent was common in all forms of agonistic behavior. Boxing, tumbling, and chasing were
seen during the fights. The frequency of these
events, as well as their form differed between species, with boxing being the most common in C.
harrisi and C. leucurus, and tumbling in C. lateralis. Among individuals where dominance was
established, threat postures included many of the
same postural components as those of the initial
encounters. Freeze-dried, postured models of C.
mexicanus were presented to captive and wild
populations of this species. The models elicited
postured approach and inspection. (Supported by
a grant from the University of Arkansas Graduate
School, a grant-in-aid from Sigma Xi. and NSF
Grant GB-3462.)
123
F. C. ELOFF, University of Pretoria.
Observations on the behavior of the gemsbok
(Oryx gazella). (Introduced by Edwin M. Banks)
Observations on the behavior of the gemsbok
were carried out in the Kalahari Gemsbok Park
(South Africa). The gemsbok is a gregarious animal, found mainly in desert or semi-desert country. Sex-ratio records show that amongst the adults
females predominate. There is no definite breeding season and production of young occurs all the
year round. The young are either hidden by the
mother or are left in nurseries with a single individual in charge. The gemsbok seems to be largely
independent of water, subsisting on a variety of
plants when water is not available. The animal is
a great wanderer and its movements are influenced
by a complex of factors, in particular its craving
for mineral licks. The chief predators are the hyena and the lion. The latter has developed a special hunting technique to avoid injury from the lethal horns of the gemsbok.
124
J. P. KENNEDY, The University of Texas Dental
Branch.
Premating behavior of the fence lizard, Sceloporus
undulatus. (Motion picture)
Premating behavior is usually initiated by • the
215
sexually active male. In the usual sequence, the
approach of the male to a nearby female is made
in short runs and momentary stops during which
the male elevates and then abruptly lowers the
body on the forelegs with a rhythmic vertical bobbing of the head. The trunk is laterally compressed and the gular region is extended, thereby
exposing the bright blue coloration of these ventral areas. This form of bobbing and display are
clearly male and may function in sex recognition.
Premating contact includes nipping. The male nips
at the base of the tail, hind quarters or lateral aspect of the female's trunk. During this time the
female may move only a few inches as the male
follows nipping at her whenever possible.
The breeding system in this population is promiscuous and not all premating behavior results in
mating. The non-receptive female responds to the
presence of a male intent on mating by elevating
her body on all four legs and hopping in short
jerky movements. Her trunk is notably arched; the
gular region is extended and the tail is elevated.
The presence of a male could be predicted upon
the basis of a gravid female eliciting the "female
hop." These sequences and the lack of heterospecific antagonistic behavior between fence lizards and
several coexisting lizards in the Big Thicket of Texas are illustrated in this 16 mm color field movie.
(Supported in part by N.S.F. Grant GU-482-A.)
125
BARBARA F. BROCKWAY, The Ohio State University.
Interactions among male courtship warbling, photoperiodic and experiential factors in stimulating
the reproductive activity of female budgerigars
(Melopsittacus undulatus).
Male Soft Warble, but not Loud Warble, stimulates the ovarian activity of females housed without
nestboxes when caged in continual darkness (Anim.
Behav. 13:575-578, 1965). No vocalizations are effective when females without nestboxes are exposed
to 12-hour daily photoperiods (Brockway, MS). A
new courtship sound, LPSC-Warble, has recently
been discovered. Over 200 females were used to
further study the reproductive efficacy of LPSCWarble, Loud Warble and Soft Warble. Taped
solos or quartets were played for 6 or 12 hours
daily for 21 days to individually-caged females with
or without prior breeding experience and kept
either in continual darkness without nestboxes (reconditions) or under 12-hour daily photoperiods
with nestboxes (L-conditions). Controls heard
either noise or noncourtship vocalizations.
Results indicate that, all else being equal: (1)
experiential factors do not affect the ovarian response of females under D-conditions but past
breeding experience potentiates the response of females under L-conditions; (2) quartets were never
more effective than solos; (3) both 12 and 6 hours
of daily vocal stimulation equally affected the
ovarian activity of females under D-conditions, but
12-hour amounts promoted more ovarian activity
and nestbox-oriented behavior in females under
L-conditions; and (4) although Soft Warble promoted more ovarian activity in females under Dconditions, LPSC-Warble promoted more ovarian
216
AMERICAN SOCIETY OF ZOOLOGISTS
activity and nestbox-oriented behavior in females
under L-conditions. (Supported by N.S.F. Grant
GB-319K)
126
LARRY C. HOLCOMB, Ohio Agricultural Research and Development Center.
Egg turning behavior of color-marked eggs in birds.
Eggs were numbered with fingernail polish in 28
species. Eggs were manipulated so that all the
numbers faced up one day and faced down the
next in 12 species. The frequency of those found
up or down on the following day was recorded.
Nine of the 12 species where manipulations occurred did not turn the eggs at random. Instead,
Chi-square values show that the numbered side of
the egg was turned down. Furthermore, Chi-square
values showed that the same nine species turned
their egg numbers down when left up more often
than they turned them up when left down.
Species that are frequently parasitized by cowbirds reacted to the colored eggs in the same manner as infrequently parasitized species. The colored
numbers apparently acted as a "disturbing" factor
to the incubating parent but they did not differ
enough from the original color to cause ejection or
desertion.
127
C. RICHARD TERMAN, College of William and
Mary.
Frequency of pregnancy failure in female Peromyscus maniculatus bairdii related to parity and social environment.
Vaginal smears of nulliparous and recently parous female Prairie Deermice were taken twice daily
for five days or until insemination (20 sperm per
low power field). Young of the parous females were
removed at birth and smearing occurred during
the first post-partum estrus. Following insemination, each female was subjected to one of the following treatments during a period from 24-72
hours after insemination: (a) stud male removed
and female left alone, (b) stud male left with female, (c) stud male removed and strange male
placed with female and (d) strange male placed
with female and stud male. All females were killed
at 10 days post-insemination and examined for embryos.
The data indicate significantly greater frequency
of pregnancy among parous females than nulliparous (.005 >P) and decreasing pregnancy proportions in both parous and nulliparous females in
the treatment sequences of (b), (a), (d) and (c).
(Supported by Grant MH-08289 from the U.S.
P.H.S.)
128
EDWARD PRICE, N. Y. State Univ. College of
Forestry.
Differential reactivity of wild and semi-domestic
deermice, Peromyscus maniculatus bairdii, to a
novel environment: effect on food consumption.
It was hypothesized that following seventeen
years of laboratory breeding (approximately 20-25
generations), a semi-domestic stock of deermice,
Peromyscus maniculatus bairdii, would show decreased reactivity (sensitivity, responsivity) to being
placed in an unfamiliar environment (activity
wheel). Two hundred subjects were used, including
the semi-domestic stock and offspring of a representative sampling of wild-caught animals. Food
consumption, body weight and activity were measured daily for five days. The effect of early environmental experience upon reactivity to a novel
environment was determined by fostering neonates
on mothers of the opposite strain (maternal influence) and offering five weeks experience in a seminatural outdoor enclosure in contrast to the laboratory (place of rearing influence). A control group
for isolation and handling was employed for both
strains.
Food consumption for all wild experimental
groups was significantly depressed (P^.01) during
the first 48 hours in the novel environment while
no change in food consumption was observed for
the semi-domestic strain. No strain differential
change in either body weight or activity was observed during the test period. Fostering had no
effect on the behavior of either strain. Semi-domestic mice given rearing experience in the outdoor
enclosure consumed significantly less (P<.01) food
than the other domestic groups but did not exhibit
the initial depression characteristic of the wild
strain. Neither handling nor isolation significantly
influenced the results obtained. The differential
reactivity of the wild and semi-domestic strains to
an unfamiliar environment may be due to genetic
changes resulting from a relaxation of natural selection and selection pressures accompanying domestication. (Supported by Grant M-5643 from the
U.S.P.H.S.)
129
ROBERT S. SCHMIDT, Stritch School of Medicine, Loyola University.
Preoptic activation of frog mating behavior.
Lesion studies show that the ventral magnocellular area of the posterior preoptic nucleus is required for mating calling to occur. The dorsal magnocellular area is necessary for orientation of the
male to the female. Electrical stimulation of the
preoptic area will elicit mating calling. Electrode
holders will be illustrated and described for both
unrestrained chronic animals and for restrained
acute preparations. (Supported by Grants NBO6673 and K3NB9513 from the U.S.P.H.S. and Grant
GB4612 from the N.S.F.)
130
BEDFORD M. VESTAL and JOHN A. KING,
Michigan State University.
Relation of age of eye opening to first optokinetic
response in four taxa of Peromyscus.
The relationship between visual development and
age of eyelid separation was studied in Peromyscus
maniculatus bairdii, m. gracilis, polionotus, and
eremicus. The maniculatus subspecies had been
selectively bred for early and late age of eye opening. Mice were tested in a transparent chamber
ABSTRACTS
suspended within a rotating drum, lined with alternating y^ inch black and white vertical stripes. Beginning within eight hours of eye opening, each
mouse was given 5-15 second trials daily until it
responded. Criterion of vision was an optokinetic
response in 2 of 5 trials. Of approximately 80 mice
tested, only the early selected line of bairdii failed
to respond within one day after eye opening. These
results indicate that mice can see conspicuous environmental features at age of eyelid separation.
The delayed optokinetic response in the early selected bairdii line, requiring approximately two additional days of maturation, indicates the possible
independence of eyelid separation and visual development. (Supported by N.I.H. Grant MH-05643.)
131
ROBERT BOICE and CAROL BOICE, University
of New Mexico.
Drinking behaviors of newly captured rodents.
Chew (Physiological Mammology, Academic
Press, pp. 43-178, 1965) suggests that captivity may
bring about genetic and/or ecotypic (e.g., stress)
changes in the processes involved in the water balance of a mammal. This study was concerned with
the voluntary and time restricted water intake of
six species of newly captured wild rodents (Rattus
norvegicus; Onychomys leucogaster; Neotoma albigula, micropus, and mexicana; and Dipodomys ordii). The results indicated that: 1. Wild Rattus norvegicus did not necessarily drink more than their
domestic counterparts or taper off in water consumption with habituation to the laboratory. 2.
The water consumption of Neotoma increased over
days and appeared to be inversely related to the
dryness of the natural habitat. 3. Dipodomys ordii
showed small but stable levels of voluntary consumption when placed on dry lab chow and came
to show eager anticipatory behaviors when placed
on one hour a day availability of water.
132
M. W. FOX, Galesburg State Research Hospital,
Galesburg, Illinois.
Multidiscipline studies on the postnatal development of the CNS and behavior of the dog.
In order to present a concept which is considered
highly significant as a natural ontogenetic phenomenon, multidisciplinary studies on the postnatal development of the CNS and behavior in the dog are
described. The close chronological correlation between EEG and EP development, myelinization and
reflex development indicates that a period of integration of component parameters of the nervous
system, in terms of perception and locomotion, underlies the onset of the critical period of socialization (Scott, J. P. (1962). Science, 138, 949-958) and
possibly of imprinting (Sluckin, W. (1965). Imprinting and Early Learning, Aldine Publishing
Co., Chicago) in precocial animals. This integration, which results in improved perceptual and locomotor abilities, allows the organism greater reaction and interaction with the environmental
milieu, thus more complex genotype-environment
interaction ensues (e.g., individual differences, nerv-
217
ous typology or 'character' emerge). Similar 'internal' and 'external' integrative processes may take
place during other behaviorally and physiologically
critical periods such as the immediate ante-natal
period and at the onset of sexual and maternal behavior. This concept is discussed in relation to
Schneirla's approach/withdrawal theory (Schneirla.
T. C. 1965). In: Advances in the Study of Animal
Behavior, 1) and Anokhin's concept of heterochronous development of the CNS (Anokhin, P. K.
(1964). In: Progress in Brain Research, 9, 55).
133
ROBERT G. WOLK, Adelphi University.
Comparative morphology of the retina of the Black
Skimmer, Rynchops nigra (Aves).
This species feeds in a manner unlike that of any
bird and is the only bird with a vertically-slit pupil. In connection with studies of its unique feeding behavior, investigations of the functional mor
phology of the eye were undertaken.
The Black Skimmer, which feeds during both
night and day, has a largely scotopic retina. Yet its
retina permits a degree of photopic vision also.
Rods outnumber cones at a ratio of 8:1. The outer
and inner nuclear layers are of approximately equal
thickness revealing an intermediate degree of adaptation to scotopic vision. The ganglion cell layer is
relatively thin.
The visual acuity of the eye is limited. When
the Black Skimmer forages during periods of low
illumination, it utilizes scotopic vision; because it
does not perceive its prey visually, this is not a disadvantage.
Comparison is made to closely-related species of
Laridae and other crepuscular and nocturnal forms.
(Supported by Grant GE-6267 from the N.S.F.)
134
A. ULRIC MOORE, R. H. BARNES, G. W.
POND, and I. M. REID, Cornell University.
Behavioral characteristics of adult pigs after recovery from early malnutrition.
A number of experiments, involving more than
50 pigs, show that several behavioral differences
exist between normal adult pigs and experimental
pigs fed a diet severely deficient in protein for eight
weeks immediately after early weaning.
A battery of tests was used to assay to what extent results were due to changed emotionality, motivation, or more limited mental capacity: 1) simple
operant food learing with fixed ratios as high as
twelve; 2) conditional emotional response based
on operant training; 3) extinction of the conditional
response; 4)dassic avoidance conditioning; 5) 48hour records of ad libitum food consumption patterns.
The experimental pigs learned the simple operant and classic conditioning more slowly than the
controls. They also developed the conditional emotional response more irregularly, but once established, it was almost impossible to extinguish the
response. When first put on ad libitum feeding,
the experimental pigs ate more and fed more often
than the controls, particularly at night.