RESEARCH PROJECT - SECRETION OF ANTERIOR PITUITARY HORMONES AND OVULATION IN SMALL RUMINANTS

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• Collection: General CIA Records
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• Release Decision: RIPPUB
• Original Classification: C
• Document Page Count: 19
• Document Creation Date: December 27, 2016
• Document Release Date: April 18, 2014
• Sequence Number: 1
• Publication Date: May 1, 1964
• Content Type: REPORT

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INFORMATION REPORT INFORMATIOW REPORT
CENTRAL INTELLIGENCE AGENCY
50X1-HUM
This material contains information affecting the National Defense of the United States within the meaning of the Espionage Laws, Title
18, U.S.C. Secs. 793 and 794, the transmission or revelation of which in any manner to an unauthorized person is prohibited by law.
C-0-N-F-I-D-E-N-T-I-A-L
50X1-HUM
COUNTRY Poland '
SUBJECT Research Project - Secretion of Anterior
Pituitary Hormones and Ovul tion 'n Small
:Ruminants
REFERENCES
DATE OF
INFO.
PLACE &
DATE ACQ
4.3
REPORT
DATE DISTR. I may 64
NO. PAGES
1N-Q--cc-LJ
50X1-HUM
THIS IS UNEVALUATED INFORMATION
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an annual report (16 pages, English) of a
Polish research project entitled "Secretions of Anterior Pituitary
Hormones and Ovulation in Small Ruminants" which is being conducted
at the Institute of Animal Physiology and Nutrition at Warsaw, under
the direction of Eugeniusz Domanski. CONFIDENTIAg
-end-,
C-0-N-F-I-D-E-N-T-I-A-L
STATE
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I AIR
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CROUP 1
Excluded from automatic
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declassification
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INFORMATION
INFORMATION REPORT INFORMATION REPORT
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? 07,0A.V
C
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? Institute of Animal Physiology and Nutxition
(. Polish Academy of. Science )
Jab/onna near Warsaw
'Poland
Chief. Inve tigator
Title of problem
eniuse DOWSI
oretion Of anterior pituitary
hormone? and ovulation in small
ruminants .0
50X1 -HUM
(GAmi PIMPL
. ? GROUP 1
Excluded from automatic
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Summary.
The aim of our work is to investigate the role of the hypotha?
lamus and the process of transmission, from the hypothalamus to
the hypophysis, of the impulses which regulate the secretion and
release of gonadotropins by adenohypophysis An sheep. ?In pre?
vious GMC experiments on this problem we investigated the role
of the pituitary stalk in the process of impuls?transmission
using the method of the stalk section. In these experiments
we were able to evidence in sheep after stalk section the di?
sappearence of ovulation as well as the atrophic changes in
adenohypophysis and in sexual organs'. We Showed also that the '
development of the mammary gland for lactation as Well as the
secretion of milk were Strictly bound with the integrity of the
hypothalamo?hypophyseal junction. Thus on the basis of these
results we were able to conclude that the hypothalamus and the
hypothalamo?hypophyseal junction in sheep plays a fundamental
and controlling role in the secretion and release of gonadotro?
pio hormones (FSH, LH, LTH).
Starting from the thesis that this controlling role is exerted
by neuroseoretory materials elaborated in oentral nervous sys?
tem (CNS) and transmitted on the vascular way of the pituitary
stalk, we have undertaken in 1963 the search for these neuro?
secretory substances responsible for the release of gonad's:Aro?
pie hormones by adenohypophysis. For these purposes we prepa?
red extraots from various parts of the hypothalamus and other
parts of CNS on the chemical way and we tested their activity
by biological methods. Namely, we injected these extracts imme?
diately into the hypophyses in rabbits and sheep (the method
in vitro) or we added extracts to the incubated slices of adeno?
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hypophyses and after incubation we titrated the content og gona-
dotropic hormones in incubated medium (the method in vitro).
By using the method of injections of extracts immediately into
the adenohypophysis in rabbits we induced ovulation in these
animals.
By the method in vitro we were able also to show the stimula?
tion of release of gonadotropins by the slices of adenohypophy?
sis.
Thus, our experiments seem to show the presence in neural tis?
sue of median eminence of some substance(s) being able to pro?
mote the release of gonadotropic hormones by adenohypophysis.
These results raised the question as to whether this substance,
obtained by extraction from nervous tissue, releases FSH and
LH together or LH only. The experiments concerning this ques?
tion are currently being performed.
In 1.963 we continued also our work on vascularization of the
pituitary gland. For in the physiology of this gland and espe?
cially in investigating the process of impuls transmission from
the hypothalamus to the hypophysis an understanding of the struc-
ture and architecture of the vascular system of the hypophysis
is indispensable. As important achievement of these investiga?
tions and worthwile to note there was a finding of hypothalap.
mopetal flow of venous blood from adenohypophysis. This con?
cept of hypothalamopetal flow of venous blood from adenohypo?
physis, gives rise to the idea of the feed?back regulation pos?
sibly existing between CNS and adenohypophysis. And this idea
has not been yet a subject of any investigations.
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1.o/eover in 1963 the technical preparatory work to investiga-
tions on the function of the hypothalamus in sheep was being
performed. For these purposes we prepared histological slices
of the hypothalamo-hypophyseal area of the brain of sheep to
design the stereotaxioal map of this area. By means of coordi-
nates oftstereotaxical map we will be able to place lesions at
Particular areas in hypothalamus and on this way to localize
the function of these areas.
II. Detailed report.
Introduction.
As presented in our first and second annual report, the main
aim of our work was to investigate the role of the hypothalamus
and the process of transmission, from the hypothalamus to the
hypophysis, of the impulses which regulate the secretion and
release of pituitary gonadotropins, and, especially, of FSE
and LTE in sheep. Starting from the generally accepted thesis
that the nervous regulation of adenohypophysis is exerted by
neurosecretory materials transmitted from the median eminenoe
to this gland on the vascular way of the pituitary stalk, we
investigated first the role of the pituitary stalk in the pro-
cess of this transmission using the method of stalk seotion.
By this method we were able to evidence that the ewes after
pituitary stalk section showed no sexual cycles, ovulation had
disappeared and the autopsy of these animals revealed atrophy
of ovaries and uteruses. Histological examinations of these
pituitaries, stained with heamatoxylin . eosin, and according
to PAS method, showed in all animals with the stalk section
an atrophy of
the glandular tissue and a hyperplasia of conneo-
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(OtN9C111:0"
tive tissue. The dimensions of cell cytoplasm and nuclei were
reduced, delta cells showed no granulation ti:e. were PAS nega-
tive).
Thus on the basis of the above cited findings we were able to
conclude that the disappearance of ovulation as well as the
changes in pituitaries and in sexual organs were caused by the
lack of impulses from the hypothalamus to the adenohypophysis
ID following the transection of the pituitary stalk.
?
Moreover we investigated also the role of the hypothalamo-hypo-
physeal junction on the process.of lactation. And we were able
to show that the development of the mammary.gland for lactation
as well as the secretion of milk were strictly bound with the
integrity of the hypothalamo-hypophyseal junction. Vhen this
junction was damaged, the mammogenic and lactogenic function
of the pituitary gland was abolished. After th,pse facto had
been stated the following experiments were continued and under-
taken in the third year of our investigations: ?
1. Experiments searohing for the substance(s) in the hypothala-
* mus responsible for the release of pituitary hormones (1H
and FSH).
2. Histological investigations upon the vascularization of the
pituitary gland in sheep.
3. Teohnioal preparatory work to investigations on the function
of hypothalamus by the method of lesions.
cogr
L rktiv t,i .
tic.3..0 W.4.
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Progress Report.
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Experiments searching for the
substance(s), in the hypothalamus
responsible for the release ef
pituitary gonadetropins.
To investigate this problem we prepared extracts from various
parts of the hypothalamus and from the cortex, we infused them
immediately into the adenohypophysis of rabbits and sheep and
observed the reaction of the animals after these infusions.
These extracts were tested also by the method in vitro; namely,
they were added to the incubated slices of adenohypophyses and
then the relwased gonadotropins in incubated medium were titra?
ted. The extracts from nerve tissue were prepared in the fol?
lowing way: The whole brains were removed and deep?frozen im
mediately after the slaughter of the animals. Then all the
isolated median eminences were put together and all the anterior
parts of the hypothalamuses were also put together. Isolated
tissues were extracted with 0,1 N HC1. This crude extract was
lyophilised, stored in the refrigerator, and immediately before
testing it was dissolved in saline. Extracts of cerebral cortex
(from the frontal lobe) were prepared in a similar way and
used for eontrol infusions. The lyophilised crude extracts were
dissolved immediately before tests in various concentrations
in physiological saline and slowly infused into the adenohypo.
physis of rabbits and sheep through chronically implanted can?
nulae. In rabbits the dose level amounted 25 AA and eentai?
nerd as low as 0.175 mg equivalent (concentration 1) and as
(f):
t
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(C'r".'" 6
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high as 1.4 mg equivalent (ooncentration II) of lyophilized
tissue per one animal. Ovaries in rabbits were examined by
.1aparatomy after 48 hrs and on the 5th day after infusion.
The results of these infusions were as follows:
Table ?4.
Part of the brain No. of
and concentr. of rabbits
lie extract used for
test
.111.411??????M ????????????????????????????????41M.
Changes in ovaries afterinfusion
No of rabbits
with bloody
spots in ova-
ries 48 hrs
after infusion
No of rabbits No of corpora
with aorpora lutea per one
lutea, 5 days animal
after infusion
median emin.
rabbit cone. I.
median emin.
sheep, conc. I.
median emin.
rabbit conc. II.
median emin.
sheep cone. II;
hy2ipaalamus
conc. II.
cerebral cortex
rabbit conc. II.
Pitocin /oytocin/
0,4 I.U. /Parke
Davis!.
Pitressin /vaso-
pressin/
2.0 I.U. /Parke
Davis!
5
a
9
3
5
6
3
3
acetyl choline Oa 3
?????????????????=marwra....???????????? ???????????? IMP.O? 01M.?
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8
3
2 -
1
0
6
8
2
7
1
9
4111111.
0,11.
In sheep, we have to data been able to carry out only a few
experiments with injections of extracts. We injected these
extracts into two sheep during the sexual cycles., in one on
???-?t1 A
rkr'...
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CORDRIPAL
the 10-12 and into the second on day of he cycle.
The dose level amounted 200A-1 and contained as much as 14 mg
of lyophilized median eminence of sheep. The effect of the
injeotions in sheep was tested by vaginal smears and by be
running of the ewe with the ram. To date we received positive
reaction i.e ovulation in one sheep only, namely
the sheep
wMch.was injected on he l2 day of its sexual cycle.
2. !nye igati.ons upon the vasott?
riza 'Oil of th
in sheep..
a
p ituitary gland
An understanding of the structure and arohi eotu e of the vas-
cular system of the hypophysis is indispensable in investiga,
ting the physiology of this gland and especially in investiga-
ting the process of impuls transmission from hypothalamus to
the adenohypophysis. This basal problem of blood ciroulation
in hypophysis and especially vascular relations between hypo-
thalamus and hypophyais in sheep is known .rather very littlE,
therefore we undertook this
To this aim the arteries an the. veins f the heads of sheep
were under pressure injected with indian ink or prussian blue,
then after their fixation in formalin? the hypothalamo- po-
phystal areas of the brain were embedded in oelluloidin and
sectioned in slices fifty micron thick. All slices were exa-
mined in the stereoscopic microscope and on the basis of these
examinations the vascularization of the pituitary gland and
the blood circulation-relation between hypothalamus and hypo-
COVIFIDNI14,
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(Melina
physis were reconstructed. The pictures Nr 3 and 4 represent
these reconstructions. According to these pictures the ante?
rior and bosterior lobes receive separate arterial supplies.
Namely, the anterior lobe (adenohypophysis) is supplied by the
arteries running from circulus of Willis and approaching to the
median aminence. Branches of these arteries encircle the me?
dian aminence like a ring. Numerous small descending branches
spring from this ring and form the capillary bed of the stalk.
Capillaries around the stalk were seen to empty into the dila
ted channels of the anterior lobe. The effluent blood drains
from adenohypophysis through two venous systems. The first
one drains the blood directly into the venous sinus which sur?
rounds the pituitary gland (in pituitary fossa) and the second
one conveys the venous blood upward to the floor of infundibu?
lar recess, where the veins break up into a system of fine
channels, a secondary distributing net.
According to the anat mical sizes of veins of these two eye?
we suppose that the majority of the blood is drained
from the adenohypophysis upward hypothalamopetal and smaller
part of its is conveyed directly into the perihypophyseal
sinus.
The post eror part of the pituitary gland is euppli.ed by the
arteries originating from the so called rate situated in pi?
tuitary fossa and formed by the internal carotid arteries.
Connections between these two arterial ssteme of anterior
and, posterior part of the hypophysis, exist in special anato?
mical places of the gland; they are, however, not numerous
(see picture Nr. 4)
(ONFIDENTIAL
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(.1jVir 111) V4 UAL.
The effluent blood from posterior part of the hypothalamus is
drained also by two ways, namely by the immediate way into
perihypophyseal sinus, and by the veins running upward hypo-
thalamopetal.
3. Technical preparatory work to
investigations on the function
of hypothalamus by the method
of lesions.
This work was carried out to prepare stereotaxical map of the
hypothalamo-hypophyseal area of the brain in sheep. This map
is indispensable for the method of lesions and other methods
applied in electrophysiological investigations of CNS. For
these purposes the hypothalamo-hypophyseal areas of the brain
of sheep were embedded in celloidin and sectioned at fifty
micron thickness. Every fifth section was mounted. The sec-
tions of two brains were stained for cells and those of two
other brains were stained for fibers. On the basis of these
histological sections, coordinates for particular nuclei of
the hypothalamus will be drawn. This last cited work is cur-
rently being performed.
CONFIDENTIAL
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Tr:FIDMIAL
The abili
of extra?
o indaoe bvula
rabbits by
ha amus directly
?physis and .the dif n the action between the extrac
from par icular parts of the hypothalamus and, the cortex seem
to show the preOn in nedian emlnence
substance being able to promote the rel
by the denohypophysis. This. phenomenon ?
a? to whether, in indueed ovulation by iauion of the ex-
some tranamita4
of Sonadotr pins
*see the qUestion
d Iflax,e released t othez, or LH oily. Our pre
sful attempts to induce ovulation in sheep
'With, these extracts
does not 0
sted iniri
em to dhow that the 1oaeix of FSH
g extracts
atted by
to disou
with Dr.
e
The
scientists nOBt advanced ta.
d Dr.S.LLoCannt
ober and Nov tb ex' 1963).
The result of
th above cited
'date this probl in further
tted

wyc Departnen
DOPar
of Penney
hedio
rnia
$ Sohool of
lphia.
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11 ?
YP0PhYse la ization
The most important pl?Oblem in the hypophyseal vascularisatio
is the relation in the blood circulation between hypophysis and
hypothalamus. Our findings of arterial blood supply of adeno
hypophysis in"sheep,by the capillary bed of the stalk is in a
whole agreement with the generally accepted concept on the by
pophyseal 'portal system. The direction of the flow Of arte?
rial blood in this system (from infundibulum to adenohypophy?
sis) supports the thesis of neurohormones transport from hyp
thalamus to the adenohypoyhyeis on the vascular way.
On the opposite,
iews on he draining of venous blood
from adenohypophysis are not conclusive. Popax as firs
in his theory of a portal circulation (dated back to 1930)
suggested the concept of.hypdthalamopetal venous blood flow
from adenohypophysis. This concept, however', was not supported
by reliable anatomic data by Popa himself and rejected later
x)
by several other. authors?x? Newer work conderning this con?
Popa, G.A
xx) Visiocki
Yewell
Barnet
U.Fi lding: A portal circulation from the
pituitary to the hypothalamus region.? J.Anat
6$, 1930.
.G.B. 1937. The vascularsupply of hypoPhysis
celebri in cat.?
D.A., 1956,. The vascular link between the poste?
rior and anterior lobes of the pituitary gland.
J.Endocrinol., 14: 24.
R.I. and R.O.Greep, 1951. The direction of flow
in blood vessels of the infundibular stalk.
Science, 113:185.
(Ot\IFIDENVAL
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oept give also little informationx) . Oar findings in sheep
support the oonoept of hypothalamopetal flow of Venous blood
from adenohypqihysis and demonstzate the breaking up of the
veins running hypothalamopetal into a secondary distributing
net in the area of infundibulum. This concept of thalamo-
pet l flow of venous nlood from adenohypophysis gives rise to
the idea of the feed-back regulation possibly existing bet-
ween CNS and adenohypophysis, idea being not yet a subjeot of
any investigations. For these reasons our investigations upon
vascularization of hypophysis aroused a great interest and vi-
vid discussion with several scientists+) during my-above cited
stag in the United States'.
This diseussion, however,revealed that to support the concept
of 'hypothalamopetal flow of venous blood from adenohypopkyeis,
further experiments are needed.
Moreover there is another problem very Important conneoted
with the vascularization of the adenohypophysis; namely, the
problem of arterial blood supply to adenohypophysis after the
stalk section. According to the investigations of Daniel and
Landsmeer J.F.M. 1963, A survey of the analysis of hypophy-
. sealvascularity.- Advances it Neuroendocri-
nology, University of Illinois Press - Urbana
Dr. E.T.Clegg, Ani al Husbandry Department, University of
California, Davis.-
Dr. Ganong Department of Physiology, School of Medicine
University of San Francisco,-
.
D . dc Groot and Dr. B.Donovan, Department of Anatomy,
Medical School, University of San Franoi o.
0014FIDE4TIAL
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?richardx) and supported by our (described above) the adenohy?
popkisis in sheep is supplied mainly bl arterial vessels of the
stalk. ?aniel and .1.ichardx7) were able to find massive in.
fftetions, necrosis and regeLeration in Ldenohypophysis after
the istalk section in sheep. These chanios in adenohypophysis
after stalk section were intcrpretod b aniel and Prichard as
the result of the deprivation of the blood supply to the gland.
In our expericlents none of the 6 ewes operated on and slaughte?
red about one year after operation showed7to traces of neoro?
Si8 in adenohypophysce. The proces of necrosis was alio not
foutd in 1 ewes, of which one was slautjitered 2 Q.etl."%s, and the
second one 4 weeks after the operation. Thlts, on the basis of
our observed histological ohanges, it seemed us that the atro?
phy of &landular tissue in adenohypophysis and particularly the
degeneration of delta cells after stalk section were caused
not by disturbanoes in the blood circulation but rather by look
Of functional correlation following the abaenoe of substaLces
transritting; tho irpulses from the hypothalamus to the adenoAy?
pophysis reLulating its function. This view can be supported
also Ifyy our investigations on the blood oirculation in hYPo?
physis and statement of vascular conneetions between its ante?
rior and posterior part. Fox these vascular connections give
possibilities of oonpensation in blood circulation in the adeno.
hypophwas after the o-;:;eration of stalk seotion.
411,111.11.1.1.1...????????
x) Janiel ,.,.. and 3.".trichard, r.57. The vascular arrange?
ments of the pituitary gland of. sheep.?
:yrl.Physiol.
xx) Daniel, and :'rlahard,- 1957. Anterior pituitary
necrosis in sheep produced by section of the
42:433.
Wits:
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? ? (011.i}-
Prospectus for investigations in the next year.
areamOisiONO* aeot*Oommoi.*iismemodvonui.......0".00.0e40.1.0004.40.1Ffp,...
The ability to induce ovulation in rabbits by the infusfl of
extracts from the median eminence shows the presencein this part
of the brain of some substance (factor) being 'able to promote
the release of gonadotropins by the adenohypophysis. This phe
nomenon raises the question as to Whether, In the process of ovu.
latien there is one only factor of hypothalamic origin respell ?
.sible for the release of both, FSH -and LH there are two
different factors responsible for the release of each Of these
lhormones. To elucidate this question we plan:
1. to continue our experiments with injeetions of crudeextracts
in order to induce ovulation in animals with spontaneous ovu.
lation (chiefly in sheep and also in guinea pigs)
? to fractionate our aotive crude extraots from median eminen
oe by chemioal methods and to try to induce ovulation by the
? WO of purified fractione.
eover we will damage some neural centers in the hypothalamUs
by eleotrocoagulation in sheep and we will observe the effect
of these injuries upon the functioning of the adenohypophysis.
Therefore, as it is noted above, we are now working on an histo
.-
logical map (atlas) of the hypothalamus of sheep to be used
in connection with these studies.
In addition, according to this report, we will continue our
work on the vascularization of the pituitary glaniln sheep.
CONFINNitAi
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Cho= 010 coma-
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hypothalamo tn(
Aonolmloplivocic
in Olcopt throA
!Aloi inguoionc
of =tract? aro
porforncd.
20 X 2?ay pies
ro og nocalo
?Ica trodo int
atood into cc
noUypopllycic
in rcnit for
inftvoion og
ontrcoto.
CINIF:DEVAL
Declassified in Part - Sanitized Copy Approved for Release 2014/04/18: CIA-RDP80T00246A026700300001-2
?
Declassified in Part - Sanitized Copy Approved for Release 2014/04/18 : CIA-RDP80T00246A026700306001-2
CUNtivtri
_
FRon
RETE
10 vet+ o
SIM,t5
,ornitruttio,
of pituttory.
P.:
CONFIDENtiA1 -?
Declassified in Part - Sanitized Copy Approved for Release 2014/04/18: CIA-RDP80T00246A026700300001-2