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6-1-1936
The influence of Theelin on the histology an
physiology of the primary and accesory sexual
organs of the Albino rat
Violet Alice Garrett
Atlanta University
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Recommended Citation
Garrett, Violet Alice, "The influence of Theelin on the histology an physiology of the primary and accesory sexual organs of the Albino
rat" (1936). ETD Collection for AUC Robert W. Woodruff Library. Paper 603.
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LNFI1UB~1IQE OF TRF1LIN ON T~
RISTOIJOGY AND PHThIOLOGY OF THI~
PR~L~ARy AISfl) ACCESSORY SEXUAL ORGANS
a?
TR~ AT1BI1~TO RA~
A 1~HES!S
SUBMIT~ TO 1~ FAC~ULT~f OF AT1~NtA, UNIVERSITY
IN PARTIAL 1UI~FILI~NT OF T~
~
i~ DEGREE OF ~AS1~E~R OF ARTS
DY
VIOLET ALICE GARRETT
DEPARTM~1~ OF BIOLOGY
ATILA~fl~A, GEORGIA
31~~ 1936
FOR
ii
T&~I~~ OF CONTENTS
PA~
C~AP~R
1
INTRQDUCTION
II
REVIEW OY LlTERA~I~Ii1RE
III
MATERIALS AND M&TH(I)S
I~
RESULTS
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0 C
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9
• 0 0 • 0 0*
0 0 0
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• 5 0
5.15
~zperimeflta1 Results in Females ~ 15
Experimental Results in Males 0050*0 00000000Ø• 21
Histological Picture in Females 004004400 ~ 23
Histological Picture in Males ...........~.... 24
V
CONSIDERATION OF RESULTS
V.~
Cc~CLUSIONS
VII
BIBLIOGRAPIIY
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LIST O~ TAB
?ABIE
I
PA~
F~ffeots of Theelin Injections Upon
Norma). Albino Rats
~
•O~êO
ODO
•O~
•OO~
1.1
II
Effects of Theelin Injections Upon
the Oestrus Cycle of the Albino Rat
III
Effects of Theelin Injections Upon
Eormal, Male, Albino Rats ~ 22
20
iv
LIST O~’ FIGURES
FIGURE
PAGE
1.
Oestrus Smear, Typical of normal Animal
and Animal Under Influence of Theelin .~....,.... 31
2
Metaestrus Smear, Obtained 24 Hours
After the Estbous Smear ..
a
•
a
a a a a
0
• o a
3].
a
3.
Diestrus Smear, Obtained 36 Hours
After the Estrous Smear •......................... 31
4
Proestrous Smear, 48 Hours After
the Estrous Smear . . . . .. . .
.
..
3].
...~. a... coca
33
. . .
5.
Control Uterus, Infantile and Anemic
6.
Control Uterus, A Higher Magnification
of Figure 5
. . .
•oD0OS0•e•00000000000000000S00000000q.
7.
Injected Uterus9 flyperemic and Distended
00*000 00.6
33
Injected Uterus, A Higher Magnification
of Figure 7 a a a a a a a a a a a a a e • a a a a
a o
• a a a a
33
*
9.
33
a
0 a
0 S S
Control Vagina, Closed By a Solid Cord
of Epithelium
a a
a
• a a a a
a
. a *•
• a
a~ a
a a. 00
a
a a a.., a ace
35
10.
Control Vagina, A Higher )ilagnU’ication
of Figure 9 oaeoao.aaaaa.~.0.e.*.a....a0a.ooa,.a.. 35
11.
Injected Vagina in an Experimental Oestrus
Condition (SaggitalSection) ~
12.
Injected Vagina, A Higher Magnification
of Figure 11 • . a a a a
a
a a a a a
a .
•
a a
13
a
a a
0
•
5
Injected Vagina in an Experimental Oestrus
Condition (CroesSection) 00000000.,000000
o . .
35
~.....
37
a a a
14.
Injected Vagina After Effects of Theelin
Have Worn Off 0005000aa505.OOaQaCa000000é.,5500aa. 37
15.
Immature Testis of an 18—day Old Anin~.l
16
Immature Testis, A Higher MagnifLcation
of Figure 15 •aeoeaa•aao..eaa00050000060a.000a000. 39
17.
Control Testis at Beginning of Puberty
18
Control Testis of Adi~1t Rat, 109 Da~rs Old
•a~...a,
41
19.
Injected Testis9 Animal Injected 13 eeks
With Theelin a
a a a a a a • • a o a a a a a
a a a a a a a
.
41
a • a a a a 0 a a a
a a
..~.......
...........
39
39
CR~PTER I
INTRCIDTJO TIGN
A crystalline substance was obtained from the urine
of pregnant women by Doisy and Butenandt, independently, in
1929
Doisy has given it the name, “theeli&’, which is derived
from the Greek term, “theelus”, meaning fer~1e. Its function
is the maintenance of the accessory genital organs and the
secondary sex characteristics of the female in full functional
condition0
Since males and females differ in so many essential
respects, it was natural to expect differences in the reaötion
of the two sexes to theelin0 It is the purpose of this paper
to check and attempt a verification of certain facts already
established and to compare the effects of theelin on males
and females thus:
1.
Effects of theelin injections in the i~Tmiature female
rat as related to:
(a) Production of “heat° or the ‘~oestru&’ cycle
(b) Effects on the vagina, uterus, and uterine tubes
2.
Effects of theelin injections on the immature male
rat, as related to:
(a) Development of the testes
(b) Descent of the testes into the scrotal sac
I
2
Since the principal action of theelin is to induce growth
in the tissues of the accessory genital organs of the female,
it seems natural to conclude that this same hormone was
probably the effective agent responsible for growth of these
organs which results in the attainment of puberty. The im
mature rat is especially well adapted for experimental tests
of this kind because the ~iagiaa does not open until the
attainment of puberty. Rats and mice also have the shortest
known oestrus rhythm among ra~smnals, four cc five days for
both growth. and degeneration phases. During this oestzua
period the vaginal epithelium corni~ies, pr viding a olear~
cut end point, which can be followed in the living animal.
For these reasons the irweatigator decided to use the
immature rat as a test animal.
CRAPTER II
REVIEW OF MTERATUBE
The h-ori~one, theelin, is produced in the follicles. Its
principal f
tion is its influence upon accessory- sex organs
and secondary sex characters of the female, keeping them in
full functional condition0 This was demonstrated by Allen and
Doisy, 1923, by injections of liquor folliculi from follicles
of hog ovaries. The test animals uSed were mice and rats.
Several injections of the active substance induced the accel
erated growth, byperemia, and secretion in the genital tract
characteristic of the period of oestrus. The experimentallyinduced conditions were equal in degree to the maximum attained
in the normal animal under ovarian influences. While in this
induced oeetrus condition the test animals experienced typical
mating instincts, at times taking the initiative in courtship,
and accepting coitus in a normal way
Therefore, sex instincts
are ultimately dependent upon this hormone (Allen and Doisy
and collaborators, 1924). In other rats the full oeatrus
growth reactions of the vagina may be obtained without evidence
of mating reactions (Hemmingsen, 1929).
The most important of the reactions to this substance is
an increase in growth in the accessory genital organs, vagina,
uterus, uterine tubes, and mammary glands. The epithelial
I
• .~.V
.A.~
4
tissues show a high incidence of cells in mitotio division.
The epithelium of the va€inal wall is completely replaced
at each oeetrus perio4, Secretion from glandular tissues
and a change in contraction rate and amplitude of muscular
tissues of the genital tract follow the wave of growth.1
These processes culminate in ass trus, the normal period of
se~cual activity of lower m~mnais, which can be induced
experimentally in ovariectomized animals by injections of
the ovarian foflicular hqrmono (Allen and floisy, 1924). The
temporary cessation of secretion of this hormone or a
decrease below a threshold level fails to sustain this
byperplactic condition and much of the new growth result
ing from the endocrine action degenerates and is removed,
returning the accessory genital organs to the castrate or
resting condition.2 After the hormone is exhausted, de
generative c anges set in and most of the tissue resulting
from its action during t~e preceding period is removed.
Thus the alternate absence and presence of the follicular
hormone is sufficient to supply the causative meohaniam of
the oestrus cycle (Allen and Doisy~ and collaborators,
1924.)
In immature animals transformations in the genital
tract may be induced both in the normal and in the ovariect
omized rats by injections of the ovarian follicular hormone
(Allen, Edgar and Doisy, ‘.A. 1924). Thickening of the vagina
and cornifjcatjon of the epithelium result in opening of the
ar
en,
ex an
n era
ecre one, a
ore:
4,
Wifliams and Wilkins Company, 1932
~Ibid., p. 404
5
vaginal orifice. The tiny infantile uterus begins rapid growth
4rd differentiation; besides enough secretion forms to
die
•~tend the uterus greatly.1
Unusual effect of ovarian extract was demonstrated in
the pocket gopher by Hisaw, 19~5. He was able to cause res rp
tion of the pubic bones, leaving the pelvic girdle open in
front in young females and males with injections of ovarian
extract.
The action of the ovarian extract in the immature animal
gives exactly the same histological results in the accessory
genital organs as are obtained from implants of anterior
pituitary tissue or injections of active extracts of that
gland (Smith and Engle, 1927). Implants of anterior lobe in
immature female rats and mice produce changes in structure
that are characteristic in the vagina and uterus of the nor
mal rat and mouse at puberty and. maturity, These changes are
identical to those reactions to the ovarian follicular hormone.
The conclusion then that the anterior pituitary hormone stim-.
ulates the accessory genital tissues to growth is obvious.
That the anterior pituitary substance has no direct action
upon the accessory genital organs is shown by negative results
with ovariectomized animals.2
It thus must act through the
ovaries by stimulating their endocrine activity0
Evans and Simpson, (1928), associate two distinct sub
stances with the ~~zmna1i~z~ anterior hypophysis: one a growth
~ Edgar Allen, Sex and Internal Secretions, Baltimore:
Williams and Wilkins Company, 1932~ p. 415
2 Ibid09 p. 417.
6
hormone and. the other a maturity-provoking hormone0 The
maturity-provoking hormone stimulates the development of the
gonads.
Injections of ovarian extract in normal adult females
are successful in accentuating and maintaining growth in the
accessory genital organs. Continued administration of more
than minimal doses may eliminate completely the degenerative
phase of the cycle in the vagina of the normal animal as
determined by the cell contents of the vaginal smear. The
uterine mucosa is maintained on a high functional plane)~
Injections of the follicular hormone into senescent
albino rats, after the wave of sexual activity, have resulted
in the typical wave of oestrus growth in the accessory organs
similar to that already described in ovariectomized animals
(Slonaker, 1927). Such injections have no stimulating effect
on the ovaries. There is no evidence to indicate that this
substance will rejuvenate the senescent female.
The adult male shows few distinctly harmful effects
from injections of moderate doses of ovarian follicular
hormone (Bugbee and Simond, 1926). In the inm~.ture male,
however, it is possible to inhibit the normal growth of the
spermatogenic tissues with relatively small amounts of
ovarian hormone (Goldirig and I~amirez, 1928). The testes remain
infantile, sperm are not developed, and descent of the testis
into the scrotum is inhibited. The testes can be held in this
state of development f or sever 1 weeks, well past the normal
time of attainment of puberty. Very soon after the cessation
1 Edgar Allen, Sex and Internal Secretions, Baltimoreg
Williams and Wilkins Company, .1932, pp. 419-420
7
of injections, however, the retarded testes begin to grow very
rapidly and descend into the scrotum. Two weeks after cessation
of hormone treatment the testes have recovered, from the in
-hibiting effects, normal, mature cells in ail stages of sperm-.
atogenesis are found, ~uid sperm are present in the lumen of
the seminiferous tubules. These inhibitory effects may be
explained. by a depressing aotic~ upon the anterior pituitary
as far as its production of secretion essential to nermal
growth and function of the testes is concerned.
Doisy, Clement, Veler, and Tha~er, (l9Z~O), reported the
isolation of the crystalline follicular hormone before a meet
ing of The thirteenth International ~h~siologica1 Congress at
Boston, August 1923. Upon further study they found that in
the preparation of the hormone, two crystalline forms could
be obtained~ one, a leaflet type of ãrystal, appeared when a
solution of the, hormone was precipitated with water. The
injection of solutions of the crystals produces the oestrus
response of ovari.ectomiaed rats.
K~md.~, D’Armour, Qarlson, and Gustaveon, (1930), reported
that injections of the female sex hormone cause secretory
activity of the uterus, swelling of the vulva, hemnrragic dis
charge from the vagina, and accentuation of nmti.ng ins~tincts
in the dog~
Aocordir~g to ~utez4a~dt and 1~rrian, (1931), Doisy and
his fellow werke~s reported the isolation of a crystailine
substance out of prognancy ur~ine that was identical with one
they reported in 1929
There are two active substances in
pregnancy urine; their formulas are C18H~O2 (called Theelin
8
by I~oiay,
di-bydroxy Cestrin by ~arrian, and Progynon by
~utendanêt
)
and C~H~O3 (called Theelol by Doisy, tn
~ydroxy Oeetrin by Marrian, and Hormone Hydrate by Butendandt)0
Theelin is a ~ydrate of ~heelol/
The exact nature of the action of theelin upon the genital
tissues is still not understood0 “Since we cOnsider hormones
to be metabolic products of cells, secreted directly into the
blood stream or tissue fkiids, it seems reaBonable to look for
action directly upon the cells which are invol~od. It seems
possible that such action may be centered on the mechanism
regulating the blood supply0 There is little doubt but that
these hormones directly stimulate the vascular control meohan
ism0 for a hyperemia attendant upon the de’~elopment of oestrus
conditions has been recorded by the earl&est investigators in
this field”.1
A normal rhythm of alternate dilation and constriction
causing blushing and blanching of tissue exists in the uterus
of iii~ymn~.ls. During oestrue this vaso-dilation occurs more often.
Markee, (1932), studied experimentally the role of ~rascular
control in theelin by transplantation of uterine mucosa to the
anterior chamber of the eye0 Injections of theelin into
inmi~ture uterine tissue transplants bring about the oestrus
type of vaso.cdilation within thirty minutes after the first
injection0 The blushing and blanching persists in a propor
tional amount to the hormone injected.
~dgar Allen, Sex and Internal Secretions, Baltimore8
Williams and Wilkins Compar~y, 1932. p. 425
CHAP~R IU
~TAT~RIALS A1W METHC~DS
One of the major events upon which the isolation of
the foilicular hormone depended was the discovery- of Aecheim
and Zondek, (1927) of the great concentration of the foflic
ular hormone existing in the urine of pregnant women. After
the isolation of theelin and theelol the vaginal smear
procedure of bio~rassay was used by many investigators. This
caused much confusion, for almost every investigator intro
duced his own ideas into the bio—assay procedure. Many
variables existed. Mice and. rats were used; some investigators
used a single injection and. others used as many as eight.
Curtis and Doisy, (1931) defined the rat unit as, ‘1the minimum
quantity that causes esthblisliment of the vagina orifice
within ten days in three out of five animals receiving the
hormone in six doses in three days, beginning when the animals
ar~ eighteen days old.’1 “Theelin is approximately twice as
active as theelol in adult spayed rats, whereas theelol is
six or seven times as active as theelin in immature female
rats,11
Using the vaginal smear method of assay on eighteen
day old rats approximately 0.16y of theelol causes opening
of the vagina within ten da~rs following the
first
injection;
.O8y of theeliri suffices. Theelol, with subcutaneous in
jections assays at least 1,500 rat units per mge Orally it
is one-third to one-half as active. With subcutaneous injec
tions of eighteen day female rats theelolassa~ys~abou~t1 H
~i i~L~
10
69000 rat units per mg.
Alien obtained best results with subcutaneous injections01
Aqueous solutions of Theelin are more effective when admin
istered in divided doses (Dodds, 1929).
The hormone, theelin, was employed in the investigations
reported in this thesis. It was obtained from the parke,
Davis and Company, which distributes it under license from
St. I~ouis University. Each lot of the product is tested and
approved by the Biochemical laboratory of St. Louis Univer~
sity before being released for sale. The hormone was received
in glaseptic ampoules of ~0 rat units per cc. Each ampoule
was diluted, when needed, with 7.2 cc. of distilled water
for injection material. This gave a potency of 3 rat units
per 1/2 cc., which was equivalent to the rat unit of Curtis
and Doisy.
The albino rats used were obtained from the Breeding
and i,aboratory Institute, 1567 Third Avenue, )Iew York City,
New York. These were inbred for several
enerations and uaed
when needed, thus insuring wziformity of material0 The
animals were kept in breeding cages made of galvonimed
iron wire netting, resting in a galvanIzed—iron pan. This
material was. used because it may be readily sterilized. Once
a week the cages were cleaned with a stiff brush, provided
with fresh nesting tissue and clean food dishes. Once a
month everything was washed, sterilized, and disinfected.
Such sanitary precautions were observed because, even with
Edgar Allen, Sex and Internal Secretions, Baltimore:
Williams and Wiikinà Company, 1932. p. 400.
U
the beet of care, rats may be infested with parasites or in
fectious diseases
The food consisted of cracked corn, rolled oats, and
lettuCe. In order to prevent vitamin diseases the lettuce was
provided daily; co&liver oil was mixed with the cracked corn
once every two weeks; and a dropping bottle containing yeast
dissolved in water was placed in each cage once a week. As rats
are dependent upon the constant availability of food for good
health, a dog biscuit or alice of toasted bread was kept iz~
their cages at all times0 A closed drinking-water bottle,
fitted with a rubber cork pierced by a glass tizbing drawn out
to a small opening, was placed in each cage • This was done be
cause water standing in an open dish in a rat cage becomes
quickly contaminated with urine and feces and, is thus unfit
for drinking.
E’rery animal was numbered, marked, and a record kept with
regard to date of birth, sex, individual number, and descrip
tionG The marking system used for identification purposes was
the clipping of hair in minute spots on either the head,
fore-limbs, hind-limbs, back, or stomach.
~he first series of experiments was begun July 5, 1934.
~imala of a single litter were kept in one cage and all cages
were kept under like conditions. Control groups were formed
from each litter and 1~ept under the same conditions as the
experimental groups
Subcutaneous injections of three rat units per 3/2 cc.
of theeli
were made in the abdominal region of the females,
• ,.•
.~•
•-.•~•
12
ice ~er day and for th~’ee successive days, beginning when
the animals were eighteon days old. Records were kept daily of
the opening of the va4ina. V~ginal smears o
each animal (es
perimental and control) wore made by sampling the contents of
the vaginal lumen twice daily for twelve sirneeseive dayw,
beginning when the vagina oponed
The varioti
identified after spreading them in a drop of water on a
types of cells were
lide (a
platinum wire was used) and staining with easin or mothyle~ne
blue
The cefle found indicated the condition of the vaginal
epitlieliutna. 1-n rats. and mice cornification of the ~a~inal
epithelium during ~est~rus provides a c1ea~ cut end point which
can be studied in the 1i’u~iug animal. The stages <f the oe~trus
(heat) oycrle are d~estrurn, prGeet
oestr~nn, and metaestrum,1
During the d~es~tru~z, interval9 the vaginal epithélium is thin
and many polymorp1~onuclear leukocytes are found in the vagInal
lumen. In the proestrum~ preparatary, the epithelium becomes
several layers thiok, proliferate, and begins to corni~y.
Leukocytee are rare. The oestrue, heat, phase is characterized
by cornifioation of the upper epithelial layers. The ngclei
no longer stain, leucocytes are absent, and only cornified
oe1ls~ are present in the smear. The metaestrum stage consists
of a sloughing off of the coruified layers, infiltration of
leucocytes, and a return of the vaginal epithelium to its
minimum0
~taximOw, GA., and. Blo.on~, W. A Textbook of His toi~.
Second Edition Philade2phia.
W B. Saunders Cmipany5, 1934. pp. 55~—353.
13
A positive smear c~onsists of non~-rnio1eated, cornified
scales which stain a bright red with
scam.1
The negative
smear consists of a few nucleated, epithelial cells and ma~
1~u~ocytes.
~iesues of the vagina, uterus, and uterine tubes were
removed from (1) the eighteen day old rat, before injections
were started, (2) from experimental animals at opening of the
vagina, (3) from controls at normal opening of the vagina,
(4) from e~erimenta1 ani~le when the vagina of the controls
opened, (5) from controls at the normal opening of the vagina.
The tissues were fixed in Soujn’e fluid and stained with
1ieidenhain’s Iron~-hematoz~r1in. This stain is used in the study
of cell division and in determining the finer structures of
the nuoleu~. The method used for pr
ar~ti~n of tissues and
slides was according to G~qer.2
In the males injections of theelin wei’e also started
July 5, 1934. tnj ctions of three rat units per 1/2
CC,
were
made daily orer a perind of several weeks or several months.
Daily recorde were kept of the descent of the testes in both
experimental animals and controls. Tissues were removed from
(1) eighteen day old rats befors injections were started, (2)
from controls having descended. testos, (3) from experimental
anImals still injected with theelin when th~ testes of the
controls de~ended, (4) from e~erimen~1 animals still in
jeated weeks after the testee of the controis had descended.
~ Fidgar Allen, Sex and Internal Secretiona, Ba1timore~
Williams and Wiik~ns Com’any, l9~. p. 4O~
2 ~Lichael F. Guyer, Anma2. Microlo~, Revised Fidition,
Chicago: The Univeriity of Chicago Press, 1927.
14
The tiesues were then fixei in Bouin’a fluid and prepared
for histological examination, the same procedure being foil
as that used on the female tissues.
ed
QJLA11~R IV
RESULTS
Experimental Results in Females.~
~fl~ie first experi
ments were started July 5, 1934. There were seven females
in this litter, A.
Two were used as controls. The five
experimental animals were given injections of three rat units
per 1/2 cc. of theelin twice per day for three successive
days. The injections were made subcutaneously in the abdom
inal region. During the afternoon of the fourth day after the
firn~t injection, July 8, 1934, a congested vulva and open
vagina was noted in all experimental animals. The secretion
in the vulva was a colorless, viscous mucus evident in the
vaginal orifice for about two hours only0 After two hours
the fluid dried up, leaving a pasty substance in its place,
which was gradually rubbed away by the animal itself
Vaginal smears were positive, indicating the establishment of
a typical oestrus reaction
The vagina of the two control
animals was s~ti1l closed and innnature in appearance. The ex
perimental animals and one control were anesthetized with
ether and incisions into the abdominal cavity made, Observa
tions showed the vagina, uterus, and. uterine cornua of the
control infantile. Those of the experimental animals were
hyperezuic and considerably distended. These structures were
removed and prepared for histological examination0 The
vagina of the remaining control did. not open until July 22,
1934. The animal was thirty-five days old, the age at which
15
16
normal opening of the vagina should occur. As in the case of
the prematurely opened vaginas of the experimental animals,
the vulva was congested upon Opening and the smear was positive.
Ex~periments on litter C were started November 8, 1934.
There were four experimentals and two controls. After the
injections of theelin, the vagina of three experimentals
opened during the evening of the fourth day after the first
injection, November 11, 1935, The smears were positive. The
vagina of the fourth experimental animal did not open until the
morning of the fifth day after the first injection, and the
smear made was negative, This indicated only a partial reaction
to theelin. The investigator attributed this to a slight loss
of the injected hormone, due to the movement of the animal
when one injection was made
The vagina of the controls
opened when the animals were thirty-five days old, November 25,
1935, the smears being positive.
Experiments on litter D were started February 15, 1935.
There were five experimental animals whose vaginas opened four
days after the first injection. The vagina of one control
opened when the animal was thirty-five days old. The vagina of
the other control did not open until the animal was thirty—six
days old, March 24, 1935. The smear was positive. Normal
opening of the vagina of albino rats occurs, usually when the
animal is thirty-five days old, Rowever, no accurate standard
ization regarding age of the opening of the vagina and attain
zuent of puberty can be given. The range is between thirty-five
and ninety days (Long and Evans, 1922).
•~~-*
.~-.j ~ ~
17
TA3I& I
EF~CTS OF ~JfI~1~LI~1 IHJECTIO~3
NORMAI~ AI~B INC RATS
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I
18
In Table I is listed data on the effects of theelin
injections upon nine different litters of albino rats. About
seventy-five female rats were used. Litter sisters are grouped
together. The animals were eighteen days old when the first
injections were given. Injections of three rat units per 1/2 cc
were given for three successive days. This made a total number
of eighteen rat units received by each ~experimenta1 animal.
During the afternoon of the fourth day after the first inj Be..
tion congested vulva and open vagina were noted in animals
A—I, D-l, F-i, H-i, 1-i, and K-i.
Smears made when the vagina
opened were positive, indicating the establishment of a typical
aestrus reaction. Control animals were still sexually immature.
The vagina of controls A, B,
j?,
J, K, and ~T did not open until
the animals were thirty-five days old, two weeks after the
opening of the experimental animals. The vagina of control D
opened when the animal was fifty-six days cad, control H at
fifty days, and control M at forty-nine days of age. Since
puberty ranges from forty to ninety days this was not an
altogether unnatural phenomenon
Vaginal smears made from the control animals indicated
that they pass through a complete oestrus cycle every four
days. As the vagina opened, positive smears were obtained.
These smears indicated that the animals were passing through
oestrus phase of the cycle. Smears made twenty-four hours
later were n~gative, indicating that the animals were passing
through the metaestrus phase. On the third day, thirty-six
hours later, the smears were still negative and indicated that
the animals were passing through the dies true stage. On the
fourth day, forty-eight hours later, the smears were also
19
negative and showed that the animals were passing through the
proestrus phase. By the fifth day another positive, oestrue,
smear was obtained, thus the cycle was starting over again. In
Table 2 are listed data on the effects of theelin injections
upon the oeetrue cycle of the albino rat. Eighteen rat units
of theelin caused the vagina of immature rats to open at the
age of twenty—one days. Smears made at the opening were positive,
indicating a typical oes true response to theelin. These smears
were made twice per day until the animals were about forty-five
days old. After the oe.etrus smear, obtained when the vagina
opened, a metaeetrus smear was obtained on the second day, and
a diestrus smear on the third day. On the fourth day, instead
of getting a preestrus smear, which would have indicated that
the oeatrus rhythm was about to start over again, a diestrus
smear was still obtained, Smears for the fourth day, eighth
day, and twelfth day after the first oee true smear were all
negative. On these days the oestrue phase of the cycle should
have occured, and a positive, cornified, epithelia3. smear should
have been obtained
After the first positive smear another one
was not obtained until the animals were at least thirty—five
days old. On this day normal opening of the vagina should occur
and a typical, positive, osetrus smear should be obtained.
Smears of animals A-I B—I 1).~l ~l H—i K-I ~-.1 and N-I were all
positive. On the second day a metaes true smear was obtained, a
diestrue smear on the third day and a proestrus smear on the
fourth day. By the fifth day another oestrue smear was obtained
in the same animals. Every four days thereafter the smears
showed a return to the oestrus condition
After the first
20
TA~BI~ U
EFFECTS OF THI~ELIN INJECTIONS UPON
~flI~ OESTRUS CYC
IN TH~
ALBINO RAT
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21
positive smear, obtained at the age of twenty-one, rat .T-l did.
not show another positive smear until it was fifty days old.
Smears made every day after that indicated a typical phase of
the oestriia cycle and every four days thereafter a positive
smear was obtained.
~perimeuta1 Results in Males.-
Injections of theelin were
also started in the males July 5, 1934. Three rat units per 3/2
cc were given each experimental animal, starting when the
animals were eighteen days old, for a period of at least forty
days and not longer than one-hundred and four days.
Table 3 shows the effects of theelin injections upon the
normal, male rat. The first injection, in all cases, was given
when the animal was eighteen days old. The testes of all control
rats, except animals H-C and K-C, descended when the animals
were thirty-f our days old. The testes of rat H-C descended when
the animal was forty—five days old and that of animal K-C at
forty days. Rats A-6 and. B—5 received a total of 108 R.U. over
a period of five weeks and one day, During that time there was
evidence of descending testes and by the fourteenth day the testes
bad completely descended into the scrotal sac, Rats D-4 and
?—6 received a total of 118 R.U. over a period of six weeks;
rats H-.8 and 1—12 received a total of 273 R.U. over a period of
thirteen weeks; rat J—4 received a total of 261 R.tJ. in twelve
weeks and three days; rats K-S and M-4 received 210 R.U. in
ten weeks; rat 1~T-5 received 78 R.U. in eight weeks and tour
days. In all of these cases the testes did not descend. into the
scrotal sac as long as injections of theelin were given. Ten
days after the last injection was given evidences of the
descending testes could be determined; and by the fourteenth
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23
day they had con~1etely descended into the scrotal sac. The
size of the teates and genital organs of control animals was
twice that of injected rats.
HiBtological Pi~cture in Females.—
Sections of the uterus
and vagina of litter N represent the conditions of these organs
in. all animals. The uterus of control animal N-C is infantile0
anemic, and the walls are ceflapsed (Fig. 5)
The uterine stroma
and epithelium is undifferentiated and embryonic (Fig. 6). A few
simple tubular glands have begun to deve1~p. The lumen of the
uterus is slit—like.
The uterus of injected animal N—i is hyperemio and dis
tended with secretion (Fig. ?). Typical low, columnar, secretory
epithelial cells with nuclei arranged along the basal membrane
are present (Fig. 8). These epithelial cells have enlarged and
their cytoplasm is filled with secretory droplets, wMch are
discharged into the lumen of the uterus. The uterine blood
vessels are prominent.
~he vagina of control animal N-C is closed by a solid
cord of epithelium, four to eight cells in thickness (Fig. 9).
Leucocytes are found in and under the epithelium. A mitotic
figure is found in two or three places (Fig. 10). This indicates
that some growth is taking place.
1’he vagina of experimental animal N-I is open and
typical of an animal at its first oestrus (Fig. ii). The
vaginal epitheliuin has grown to twelve to fifteen layers in
thickness. The granular layer is cornified0 its outer two or
three layers are eloughed off into the lumeii (Fig. 12). This
layer contributes the cells of the oes true smear. The germinal
24
epitheliimi contains many mitotic figures this indicate8 that
rapid growth is taking place.
Figure 13 indicates the vagina of animal L-l after the
effects of theelin have worn off. The vagina remains open, but
the epithelium is at its minimum thicimess, two to three cells0
The epitheiial cells are of the stratified squamous type.
!~any leucocytes are found in and under the epithelial layers
(Fig. 14).
~istologicai I>ioture in Ma1ea~-
Seotions of the testes of
animal 1>-B represent the conditions of the testes of an im
mature, eighteen day old rat. The testes are infantile and
sperm are not developed0 The germinal epithelium consists of
ma~zy small, embry-onic, epithelial cells. The seminiferous
tubules have no distinct lumens (Pig. 14)
Many small epithel
ial cells, arranged radially, are present. These cells are the
future Sertoli cells. Primary- sex cells are scattered between
the small epithelial cells. Between the tubules are found broad
strands of interstitial tissue.
Figure 17 represents the condition of the testes of animal
M-C a control rat at the beginning of puberty. This animal was
thirty-four days old when the testes were removed. The germinal
epitheliuni consists of email, cuboidal cells, arranged in a
single layer in some places or several layers. A sn~1l lumen is
present in many of the seminiferous tubules. The indifferent
epithelial cells of the tubules have decreased in numbers. The
se~ cells between the epithelia]. cells have increased in numbers
and have the characteristics of spermatogon].a. The interstitial
cells are arranged in strands between the tubules.
25
Figure 18 represents the condition of an adult testis, an
imal 1-8. This animal was one-hundred nine days old when the
testes were removed. The seminiferous epitheliuin rests on the
inner surface of the basement membrane. The interstitial t1~s~e
contains inter~titial cells9 blood and lymph veseels~ thin
collagenous fibres, and fibroblasts. The interstitial cells,
scattered between the tubules are irregularly polyhedral Gr
elongated. Some cells have one nucleus, others have two nuclei
The nucleus contains chromatin granules and one or two nucleoli
The seminiferous tubules are composed of Sertoli cells and sex
cells arranged around a lumen. The Sertoli cells are slender
structures, attached perpendicularly to the basement membrane.
The nucleus is oval in shape and contains a nucleolus which is
also oval in shape. The nucleus is imbedded in cytoplasm con
taining mam~ granules. All stages of spermatogenesis are found
in the tubules. Each tubule, however, has the same combinations
of generations along its periphery. Bunches of nearly mature
spermia are seen near the lumen of the tubule to the lower
left of the figure. Whorls of mature spermia are present in
the lumen of the seminiferous tubule in the middle of Figure 18.
Figure 19 represents the condition of the testes of ex
perimental animal H-8. This animal has been injected with
theelin for thirteen weeks, receiving a total of 2’7~ R.tI. This
animal is of the same age as control animal 1-8 of Figure 18
The germinal epithelium is very thin, consisting of one or two
layers of small celIa. The seminiferous tubules havo no dietino~t
lumens. Their epithelium consists of two or three cells arranged
radially. Primary sex cells fill the remaining space • No mature
26
symrsia an fotmi. Strands of interstitial tissue are present
between the tubules.
~AWPE~R V
CONSID~R~TIOH O~ RESULTS
Injections of theelin into immature female rats result in
rapid growth of the vaginal epithelium~ which causes the open~
ing of the vagina prematurely. This occurs four days after the
first of a series of ifljections. The epithelia]. layers become
cornified, the outer two or three layers are sloughed off
into the lumen. Smears made at this time indIcate that the
vagina passes through a typical oestrus reaction, similar to a
normal animal experiencing its first oestrus. Smears of a later
date indicate that after the cornified cell stage, the epithe~.
hum is infIltrated with leucocytes. The vaginal epithehiuuz is
returned to its minimal thickness, two tt four layers. The
vagina remains in this condition until puberty is reached, when
it undergoes the charaoterist4..c cornification and degeneration
phases period~.cally, every four days~
The tiny infantile uterus, under the influence of theehin
injections, begins rapid growth and the secretion of a clear
fluid greatly dis tends it.
Thus it seems that theehin induces, in the innnature female9
the accelerated growth, hyperemia, and secretion of the genital
tract characteristic of the period of oestrus in the normal
animal at puberty.
Injections of theeli-n into immature male rats inhibits
the growth of the testes, which are retained in the abdomen.
27
28
The testes and genital organs of control animals are much
larger and the teete~ descend into the scrotum during the
experiment. The testes of injected animals remain infantile
and sperm are not developed. This state of development can be
maintained for several weeks, even months past the normal time
of the attainment of puberty. Two weeks after the cessation of
injections the retarded testes begin to grow very rapidly and
deacend into the scrotum. These glands are normal, containing
all stages of spermatogenesis.
?rom these considerations it seems that theelin tends to
accelerate growth and development of the genital organs of the
female and to retard the growth and development of the genital
organs or testes of the males.
CRAPTgR VI
aoHcI,13SIo~S
1. The injections of theelin into immature female rats
induces a condition in the genital tract similar to that of
an animal experiencing its first oestrua.
2. This result may be obtained in four days by six suc
cessive injections of three rat units each, as early as
twenty-one da~rs of age. This is fourteen days before the
normal time.
3. The uterus becomes hyperemic and distended with secre
tion.
4. Rapid growth greatly thickens the vaginal wall, causing
the formation of a heavy cornified layer
This results in the
opening of the vaginal orifice.
5. After the hormone is exhausted, degenerative changes
set in and most of the tissue resulting from its action during
the proceeding period is removed.
6. Injections of theelin into immature male rats retards
the development of the genital organs or testes.
7. The testes can be held in this state of development
for at least thirteen weeks. Fourteen days after cessation of
treatment the testes descend into the scrotum and sperm are
developed.
29
Fig 1
Oestrus smear, obtained upon opening of vagina~
Cornifned epithelial cells, nuclei don~t stain, leukocytes
absent. This smear was obtained in normal animals at the
age of 35 days and in animals under the influence of theelin.
Fig. 2
Metaestrus smear, 24 hours after the oestrue
smear, few cornified scales with stainable
, many
leukocytee. ~
-
Fig. 3
Diestreu5 smear, 36 hours after the oeetrua
smear, an occasional epithelial cell with many leukocytes
-
X~R
Fig. 4
Proe3trous smear, 48 hours after the oestrus
smear. Cornified cells with stainable nuclei, few leukocytes..
This type of smear was not again obtained in injected anima1~
until they were 35 days old. This is the age the first oestru
smear is obtained in the normal animal. ~
Lei~ compound mi~croacope was employed in ail the histo
logica studies. Objective and ocular combinations wei’e
choosen to give the magnifications indicated in the legends.
31
c~9,
0
c2OQ
Fig. 1
D
na1~
~tru
ada.
Fig. 2
Fig0 3
I
pig. 4
Fig. 5
Control: Uterus (cross section) infantile,
anemic; lumen slit—like. Rat N—C. ~
—
Fig. 6
Control: Uterus. A higher magnification of
Fig. 5. Uterine stroma and epitheliwn is undifferentiated
and embryonic. c~Y~’o
-
Fig. 7
Injected: Uterus (cross section) hyperemic
and distended with secretion. Rat N-i. xIoo
-
Fig. 8
Injected: Uterus. A higher magnification of
Fig. 7. Low, columnar, secretory epithelial cells,
cytoplasm full of secretory droplets, stroina cells spindleshaped. x’~’.+’o
-
crq
0
L
C;’
0
0
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PS
Fig. 9
Control:
orifice is closed by a
eight cells thick. The
1.7 days later. Rat N—C
-
Vagina (~a~ita1 section). The vagina
solid cord of epithej.jum, four to
vagina]. orifice does not open until
(18 days old)0 x,o~
Fig. 10
Control: Vaginae, A higher magnifjcatjo~ of a
section of Fig0 90 Jv!itotic figures9 leucocytes in and under
the epitheijum. ,‘.y~,
-
Fig, 11
Injected: Vagina (Saggitaj. section), Animal ir
an experimental oestrus Condition from injections of theej.jn,
This hormone caused rapid growth9 thickening, and cornifica...
tion of the epithejium, which resulted in the opening of the
vagina Prematurely. Rat N-1~. xi~a
-
Fig0 12
Ifljected~ Vagina, A higher magnifjc~~j0~ of a
section of Fig. i1~ Epithejjum 12 to 15 layers thick; gran~
u~ar layer is Carnified, outer 2 or 3 layers are aloughe~ off
into the lumen, epithe1j~ has many 1rd~totj~ figures, x~o
-
•
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...;-,
-~
I
~
•.
.~
V
V-I....
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V.-V
.,
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._-~•t•~~•
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/-
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-
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Fig. 9
~
Fig, 10
un
un.
Ca—
the
Fig, U
Fig. 12
Fig 13 — Injectedg Vagina (cross section). Thick wallec
epithelium, with heavy cornified layer being sloughed off ml
the lumen; a typical oestrous condition. Rat H-2, 22 days 0]
Fig. 14
Injected: Vagina (Saggitaa. section). After
effects of theeljn have worn off. Vaginal epithelium at
minimal thickness, 2 or 3 cells. ~
—
LI
~
°PtO
ovrc
P0E
Fig. l5~
Immature testis (cross section) ~.rrangement
of the seminiferous tubules. Rat D-8, 18 days old. ~‘i~io
-
Fig. 16
Immature testis. A higher magnification of’ a
section of Fig. l& Sperm not developed, seminiferous tubuleE
have no distinct lumens, primary sex cells in tubules. .x~
-
Fig. 17
Control: Section of testis (cross section) at
beginning of puberty. Small lumen in seminiferous tubules,
sex cells have appearance of spermatogonia, epithelial cells
of the tubules resemble Sertoli cells. Rat IL-C, 34 days old~
-
p.
.
0)
a
C)~
‘-I
aq
a
~w
U,
Fig. 18
Control: Section of adult testis (cross sectjo~
Seminiferous tubules have mature spermja in the lumen. All
stages of spe~rzuatogenesja in tubules. Rat 1-8, 109 days old.,
—
Fig. 19
Injected: Section of testis (cross section)
under the influence of theelin for 13 weeks, This animal is
of the same age as control animal of Fig. 18. Seminifero~
tubules have no distinct lumens; primary sex cells present,
no mature. No mature spermia nor stages of spermatogeneeis0
Rat 11-8, 109 days old. Xt~
—
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BIBLIOGRAP~1Y
Allen, Edgar A.
Bal tirnore:
Sex and Internal Secretions
The Wi fliams and Wilkins Company, 1932.
Allen, Edgar A. and Doisy,
Preliminary report on
partial purification,
Reprinted from J. Am
E.A. 1923.
An ovarian hormone.
its localization, extraction,
and action in te.st animals.
Med. Aas’n., vel, 8’l~ p. 819.
Allen9 Edgar A0, and Doisy, B .A. 1924
The induction of a
sexually mature condition in immature female rats by
injection of the ovarian foUicular hormone. Am. J0
Physiel. Vol. 69, pp. 577-588.
Allen, Edgar A., Doisy E,A., Francis, 5.?., Gibson, N.Y.,
Robertson, L L., Colgate, C.E , Kountz, W.B., and
Johnston, C.G., 1924. The hormone of the ovarian
folliclet its localization and action in test animals,
and additional points bearing upon the internal 8ecretion of the ovary. Am. J. Anat. Vol. 34, p.133.
Ascheim, S., and Zondek, B. 1927. NypopbysenvorderlaPPefl
Hormon and Ovarial-hormone im Horn von Schwangeren.
Kim. Wchnschr , Vol. 6, p~l322.
Butenandt, A., and lLarrian, G.F., 1931. Zur Kenntnis des
Kryst-a].lisiertefl FollikeIhormOris (Ovarial- odor
BrunathOrmOfl5). Ztsohr. f. Physiol. Chem,, Sonderabdruc
aus Band 200, Heft. 4-6, pp. 277-286.
Bugbee, B P., and Simond, A.B. 1926. The effects of injec
tions of ovarian follioular hormone on body growth and
sexual development of male and female rats. Endocrin.
Vol. 10, p 360.
Curtis, Jack M., and Doisy, E.A 1931. The Bioassay of Theelol.
Reprinted from Journal of Biological Chemistry, Vol. XCI,
2, pp. 647-650.
Dodda, B.C. 1929. Administration of Theelin, Biological
Abstracts, Vol. 3, p. 1741, No 18588.
1~O.
Doisy, E.A., Clement, D., Veler, and Thayer, S 1930. The
preparation of the crystalline ovarian hormone from the
urine of pregnant women. Reprinted Jour. Biol. Chein,
Vol. LX3CCVI, No. 2, pp. 499 509.
Evans, Herbert M., Simpson, Miriam, B 1928. Antagonism of
growth and sex hormones of the anterior hypophysia.
Reprinted from the Journal of the American Medi~a1
Association, Vol. 91, pp. 1337—1338.
Gelding, G T , and Ramirea, F.?. 1928. Ovarian and placental
hormone effects in normal immature albino rats.
Eadoerin. Vol. 12, p. 804.
U
Guyer, Michael ~?. Animal Micrology. Revised Edition, Chicago:
The University of Chicago Press. 1927
Remeingeen, A.M. 1929. Attempts of indicating mating instincts
in spayed rats and mice with ovarian extracts Am J.
PbyaiOl. Vol. 90, p.383.
Hisaw, I.L. 1925. The influence of the ovary on the resorp
tion of the pubic bones of the pocket gopher, Geomys
bursarius. J. Exper. Zool., Vol 42, p. 411.
Kunde, M M., D’Amour, F E~., Carleon, A.J., and Gustavson, R G.
1930 Studies on ?Aetabolism. VIU
The effect of estz’in
injections on the basal metabolism, uterine endoxaetrium,
lactation, mating, and maternal insUncte in the adult
dog. Am. J. Th~rsiol., Vol. 95, p. 630.
Markee, J.E. 1932. Phyt≥miic vascular uterine changes. Am. J
Physiol., Vol. 100, pp. 3239.
Maximow, A.A., and Bloom, W. A Textbook of Histology. Second
Edition: Philadelphia: W.B. Saunders Company, 1934.
Moore, C.R. 1930. A critique of sex hormone antagonism.
Reprinted. Proc. 2nd. [nt. Congress Sex Research, p. 293.
Slonaker, J R. 1924. The effect of the follicular hormone on
old albino rats. Am. 3. Pbysicl., Vol. 81, p. 325.
Smith, PJ. and Engle, E.T. 192’?
Experimental evidence
regarding the role of the anterior pituitary in the
development and regulation of the genital system.
Am. 3. Anat., Vol. 40, p. 159