HORMONES AND BEHAVIOR 9, 249-263 (1977)

Aromatization: Important for Sexual Differentiation of

the Neonatal Rat Brain

BRUCE S. MCEWEN, IVAN LIEBERBURG, CLAUDE CHAPTAL,

AND LEWIS C. KREY

The Rockefeller University, 1230 York Avenue,

New York, New York 10021

This paper examines the hypothesis that testosterone (T) produces its differ-
entiative effect on the neonatal rat brain after undergoing conversion in situ to
estradiol-17P (E,). We examined the abilities of an aromatizing enzyme inhibitor,
(1,4,6-androstatriene-3,J7-dione) (ATD). and an anti-estrogen, CI628, to inhibit
sexual differentiation. Male and female rats were treated during the first few days
of postnatal life with ATD or CJ628, and females were treated on the following day
with T in Silastic capsules or its propionate (TP) in oil. ATD and ATD+T females
were normal with respect to time of vaginal opening, ovarian weight, ability to
demonstrate an LH surge, and lordosis behavior. T and TP females were mas-
culinized with respect to all these measures. CJ628 and CJ628+TP females had
impaired ovarian function and intermediate lordosis quotients (LQs) compared to
controls, though they were higher in both measures than T females. ATD males
demonstrated high LQs in response to estradiol benzoate (EB) + progesterone,
comparable to those of control females. CJ628males had intermediate LQs, which
were significantly higher than those of control males. These results indicate that
ATD can substantially protect the neonatal rat brain from the differentiative
effects of exogenous or endogenous T, probably by blocking aromatization. CJ628
affords only partial protection against T and produces a weak differentiative effect
due to its own weak estrogenicity. Thus, aromatization of T in newborn rat brains
appears to be essential if sexual differentiation is to occur.

Sexual differentiation of the rat brain may be achieved in large part

through the action of estradiol-17/3 (E,) which is formed locally from
testosterone (T). Estrogens and aromatizable androgens are effective
agonists in this process when given to newborn female or castrate male
rats systemically and intracerebrally (Nadler, 1973: Hayashi and Gorski,
1974; Whalen and Rezek, 1974; Docke and Darner, 1975; Doughty,
Booth, McDonald, and Parrott, 1975a, Korenbrot, Paup, and Gorski,
1975). Moreover, the hypothalamus, preoptic area, and amygdala of the
newborn rat contain high levels of aromatizing enzyme activity (Naftolin,
Ryan, Davies, Reddy, Flares, Petro, and Kuhn, 1975).
There is also some evidence that agents which interfere with aromatiza-

249
Copyright @ 1977 by Academic Pres>. Inc.
All rights of reproduction in any form reserved. ISSN 0018-506X
2.50 McEWEN ET AL.

tion or E, action attenuate or block effects of T. For example, barbiturates

have been reported to attenuate some of the masculinizing effects of T and
may do so by competing directly with T for the hydroxylation reaction of
the aromatization enzyme complex (Arai and Got-ski, 1968; Clemens,
1974). Estrogen antagonists, such as MER 25, have also been reported to
attenuate sexual differentiation induced by T (Doughty and McDonald,
1974: Doughty, Booth, McDonald and Parrott, 1975b).
Several newer and more effective drugs provide opportunities for more
definitive testing of the role of aromatization in sexual differentiation of
the rat brain. In this paper we report the ability of a potent aromatizing
enzyme inhibitor, androst-1,4,6-triene-3,17-dione (ATD), and of an
estrogen antagonist, CI628, to attenuate the differentiative effects pro-
duced by T in newborn rats. In a complementary biochemical study
(Lieberburg, Wallach, and McEwen, 1977), we reported that ATD inhibits
in vivo cerebral aromatization of T in neonatal rats and that C1628, while
not blocking aromatization, does reduce the association of T-derived E,
with putative cell nuclear receptor sites. Portions of the present study
have appeared in abstract form (Lieberburg, Chaptal, Krey, and Mc-
Ewen, 1977).
GENERAL METHODS
Experimental subjects were housed at 23°C in enclosures in which a
normal 1ight:dark cycle (14-hrllight: lo-hr dark) was maintained. Stud
males for testing of lordosis were maintained in a reversed dark:light cycle
(14-hr dark: lo-hr light).
Ovarian histology. At the ages indicated for each experiment female
rats were ovariectomized under methoxyflurane (Metofane; Pitman-
Moore) anesthesia. The ovaries were cleansed of fat and connective
tissue, rinsed in saline, weighed to the nearest milligram, and then fixed in
10% formalin for histology. Paraffin sections of paired ovaries (10 pm
thick at intervals of 100 Frn) were stained with hematoxylin and eosin and
were scored for the average number of circumferential corporea lutea per
section. These data are presented together with ovarian weight nor-
malized to body weight as indices of ovarian function.
Lordosis behavior. At the ages indicated for each experiment, gonadec-
tomized male and female rats were given daily subcutaneous injections of
estradiol benzoate (EB) in sesame oil for 7 days (10 pg/day for females;
20 &day for males). On the day of testing they were administered 500 pg
of progesterone (P) in sesame oil 4 hr before testing with vigorous sexually
active males. Stud males were obtained from Simonsen Laboratories
(Gilroy, Calif.). After the first behavioral test estrogen therapy was con-
tinued for an additional 3 days, followed by another progesterone injec-
tion and a second test for lordosis behavior. Testing was carried out under
low illumination in arenas measuring 18x 59 cm for 20 min or until the stud
 BRAIN SEXUAL DIFFERENTIATION 251

male had mounted the experimental animal 10 times. The lordosis quo-
tient (lordoses/mounts x 100) was computed for each animal which was
mounted 20 times by the stud in the two tests.
Steroids. Estradiol benzoate, testosterone propionate, progesterone,
testosterone, androst-1,4,6-triene-3,17-dione, and cholesterol were ob-
tained from Steraloids (Wilton, N.H.). C1628 was obtained from Dr. Jerry
R. Reel, Parke Davis, Ann Arbor, Mich. Silastic tubing (0.058-in. inside
diameter: 0.077-in. outside diameter: medium-grade tubing) was pur-
chased from Dow Corning. Silastic implants of steroids were prepared as
follows. Eight-millimeter lengths of Silastic tubing were cut. One end was
sealed with Silastic adhesive and the tubing was then filled with 5 mm of
steroid and sealed at the other end with adhesive. Implants were cleaned
by soaking in absolute alcohol for 1 hr and were then kept in 1% bovine
serum albumin solution in PBS (10 mM sodium phosphate, pH 7.0; 0.9%
saline; 0.001% Merthiolate) for at least 24 hr before use.
Statistical tests. Differences between group means were tested by
Student’s t test for experiments in which there were only two groups. For
three or four experimental groups, we utilized a one-way analysis of
variance (ANOVA) and tested the significance between individual group
means by the Newman-Keuls procedure (Winer, 1962).
EXPERIMENT 1
To assess the possible role of aromatization of T in the sexual differ-
entiation of the rat brain, we attempted to block the effect of T with an
inhibitor of aromatization, ATD, which has been shown to be effective in
vitro in a placental microsomal system (Schwarzel, Kruggel, and Brodie,
1973). In this experiment we administered ATD alone or in combination
with T and observed the consequences of this treatment for the defemini-
zation of ovarian function and lordosis behavior.
Procedure
Timed pregnant females (CD strain, Charles River) delivered litters of
pups in our animal facility. Litters were reduced in size to 10 pups. On
Day 2, females were implanted subcutaneously with 0.8-cm Silastic cap-
sules containing either cholesterol (CH) or ATD. On Day 3, these animals
were given a second Silastic implant of either CH or T. There were four
groups of treated females: control (CH:CH), T (CH:T), ATD (ATD:CH),
and ATD+T (ATD:T). Hypothermia was used on both days as anesthetic,
and the wound was sealed with collodion. Following implantation, pups
were warmed on a heating pad and then were returned to their mothers.
On Day 10, all pups were anesthetized with Metofane, and the implants
were removed. Weaning occurred at 20-22 days of age. The age of vaginal
opening was recorded and is reported in Table 1. On Day 2, Male pups of
the same litters were given either cholesterol or ATD in Silastic capsules.
 TABLE I
Effect of Neonatal Treatment with T and ATD on Sexual Differentiation
in Male and Female Rats

Treatment Number Day of Body wt Ovarian wti Corpora LQ

of rats vaginal (Days 75-76) body wt luteai
opening (m&9 section

Female
CH:CH* 10 39k 1’ 253 2 5 1.21 + .08 5.9 ” .8 89 k 3
CH:T 17 12117(NO)” 306 + 6* 0.53 2 .04* 0.03 k .03* 18 +7*
ATD:CH 18 38 zk I 261 s5 1.32 2 .05 7.8 + .4 86 i 4
E ATD:T 14 41 + 2 256 2 5 1.30 2 .05 6.6 k .6 82 i 5
Male
CH 20 - 334 -+ 11 - O(O120)
ATD 18 - 330 rt_I - 90 (16/18**)

(’ Lordosis quotient. Number of rats showing lordosis response is indicated within parentheses
b CH: Cholesterol,
P Mean k SEM.
‘l NO: Vaginal opening did not occur in 12117animals.
‘I Different from CH:CH by Newman-Kerns test: P < 0.01.
** Different from CH by chi-square test: P < 0.005.
 BRAIN SEXUAL DIFFERENTIATION 253

These implants were removed on Day 10. Ovariectomy and castration

were performed when the rats had reached 75 days of age. Testing for
lordosis was done when the animals were 140 days old.
After completion of lordosis testing, four females from each of the
following neonatal treatment groups (Control; ATD:CH; CH:T; and
ATD:T) and four males each from the control and ATD groups were
treated with estradiol in an attempt to elicit a LH surge. Two days
following a primary dose of 5 pg of estradiol benzoate, a 0.5-cm Silastic
capsule containing crystalline estradiol was implanted SCin each rat under
Metofane anesthesia. Blood samples were collected by tail vein puncture
at 1200, 1500, and 1800hr the following day. The rats were restrained in a
small animal holder (Carter Electronics, Indianapolis, Ind.) for the 30-60
set necessary to collect a 200-~1 sample. Blood LH concentrations were
measured by radioimmunoassay using the Niswender No. 15 antisera
(Niswender, Midgley, Monroe, and Reichart, 1968): NIAMD-RP-1 was
used as the LH standard and results were expressed as nanograms of
NIAMD-RPl per milliter of blood. The interassay coefficient of variation
in these measurements was 14% (n=9 assays).
Results
Females receiving implants of T on Day 3 (CH:T) showed elevated body
weight compared to controls and a reduced ratio of ovarian to body
weight on Day 75 (Table 1). Ovaries from CH:T females were almost
totally devoid of corpora lutea, and vaginal opening failed to occur in 12
out of 17 animals (Table 1). In contrast, females receiving ATD on Day 2
(ATD:CH) were indistinguishable from controls in all of the above mea-
sures. Females receiving ATD on Day 2 and T on Day 3 (ATD:T) were
also indistinguishable from controls. Typical normal ovarian histology
from four ATD:T females is shown in Fig. 1.
Sixteen of eighteen males receiving ATD on Day 2 showed good lor-
,dosis behavior when mounted by stud males. Most of these animals
showed pronounced ear wiggling and some showed darting behavior in the
presence of the studs. Lordosis responses were for the most part com-
plete and were prolonged after the stud dismounted. Yet for some reason,
the stud males did not appear to be strongly interested in the ATD males
and in many cases did not achieve 10 mounts within the 20-min session.
The disinterest of the stud males was even more pronounced toward the
control males. Out of a total of 20 control males, studs made a total of only
7 mounts (with zero lordosis responses). For 18 ATD males, studs
ounted 245 times and received 220 responses. For this reason the
lordosis
P, behavior is scored in terms of the number of rats responding
Table l), and the difference between the control and ATD groups was
j;4 dged to be highly significant (P < 0.005) by means of a chi-square
analysis.
254 McEWEN ET AL.

FIG. 1. Demonstration of normal ovarian histology in four ATD:T females from Exper-
iment 1. Stain: hematoxylin and eosin. Magnification:22 X.

Estrogen-induced LH release was observed in all of the control

(CH:CH) females and in several of the ATD:T females (Fig. 2, A and C,
respectively). LH surges were not observed, however, in any of the T
females (Fig. 2B) or in control or ATD males. LH secretion in the male
rats fell throughout the collection period, presenting a pattern similar to
that shown for the CH:T female group (Fig. 2B).
 BRAIN SEXUAL DIFFERENTIATION 255

I I I I ,

@CH!T c)ATD:T

0 0 0 0
$- t+i
0 8- z 8
TIME OF DAY

FIG. 2. Temporal patterns of serum LH in individual estrogen-primed ovariectomized

ratsimplanted with a 5-mm Silastic capsule containing estradiol. Panel A:4 CH:CH rats.
Panel B: 4 CH:T rats. Panel C: 4 ATD:T rats.

Discussion
The major finding of this experiment is that ATD pretreatment of
neonatal rats blocks testosterone-induced anovulatory sterility and sup-
pression of female lordosis behavior. Presumably, these findings are due
to the ability of ATD to block the organizational effects of testosterone on
CNS structures regulating pituitary gonadotropin secretion and lordosis
behavior. ATD treatment of neonatal male rats also blocks the actions of
testicular androgens on the neural pathways regulating lordosis behavior.
On the other hand, ATD did not appear to block the action of testicular
androgens on the development of the neuroendocrine complex regulating
preovulatory LH release. On this last point, however, it should be re-
called that the male rats in this study were in excess of 150 days of age
when tested for estrogen-induced LH release. Perhaps estrogen adminis-
tration to younger ATD-treated male rats would have resulted in
, preovulatory-like patterns of LH release. In this regard, Vreeburg, van
der Vaart, and van der Schoot (1977) have reported ovulation and corpus
luteum formation in ovarian grafts in young male rats treated with ATD as
neonates. And Booth (1977) has made similar observations using another
inhibitor, androst-4-ene-3, 6, 17-trione (ADT). These same investigators
also observed vigorous lordosis behavior in ATD- and ADT-treated
males. With respect to the mechanism whereby ATD exerts feminizing
effects in male rats, it has been established that blood levels of T are not
reduced at the time that ATD is acting in the newborn male rat (Vreeburg
et al., 1977). Therefore, in agreement with its ability to block effects of
‘exogenous T in females, ATD must be acting to inhibit the effects of
endogenous T in males.
That ATD may exert its effects by interfering with the conversion of T
to estrogens is supported by another study from our laboratory (Lieber-
256 McEWENET AL.

burg et al., 1977). In that study, cell nuclear levels of [“H]E, derived from
an injection of [3H]T into j-day-old female rats were reduced by prior
implantation of Silastic capsules containing ATD on Day 3. Tissue levels
of [3H]E2 and [3H] labeled estrone were also reduced, indicating that
aromatization was affected. Tissue and cell nuclear levels of [“HIT and
3H-labeled dihydrotestosterone ([3H]DHT) were unaltered by ATD treat-
ment. ATD did not interfere with the cell nuclear retention of “H-labeled
diethylstilbestrol, indicating that it is neutral with respect to the estrogen
receptor mechanism. We have not, however, tested whether ATD will
interfere with estrogen-induced defeminization. Such studies are under-
way.
EXPERIMENT 2
ln a further attempt to assess the role of aromatization in the action of
testosterone on brain sexual differentiation in the rat, we used an estrogen
antagonist, C1628, in an attempt to block the defeminizing effects of
exogenous TP in neonatal rats.
Procedure
Female rats with lo-12 newborn female pups were purchased from
Carworth Farms (CFE strain: now Carworth is a division of Chas. River,
Wilmington, Mass.). On postnatal Day 3 (day of birth = Day l), pups
received subcutaneously in the back 50 ~1 of water or 200 pg of C1628 in
50 ~1 of water. Collodion was used to seal the injection wound against
leakage, and pups were returned to the mother. On Day 4, pups were
given 50 ~1 of sesame oil or 100 pg of testosterone propionate in 50 ~1 of
sesame oil by the same injection route. There were three treatment groups
of approximately equal size randomly distributed among the nursing
mothers: control (water, oil injection); TP-treated (water, TP): CI628 +
TP. Weaning occurred at 21-22 days of age. The age of vaginal opening
was recorded and is reported in Table 2. Ovariectomy was performed
when females were 113 days of age. Testing for lordosis behavior was
carried out when females were 139 days of age.
Results
Females treated with TP on Day 4 showed a significantly earlier age of
vaginal opening, and their ovaries on Day 113 showed significantly lower
weight, normalized to body weight, and significantly fewer corpora lutea
per histological section. CI628 did, however, significantly attenuate TP
effects on the LQ (Table 2).
Discussion
Exposure of newborn female rats to the estrogen antagonist CI628
attenuates defeminizing effects of TP on lordosis behavior but not on
 TABLE 2
Effect of Neonatal Treatment with TP and C1628 on Ovarian Function
and Lordosis Quotient

Treatment Number Day of Body wt. Ovarian wti Corpora LQ

of rats vaginal (Day 75) body wt lutea/
opening hk) section

5 Control 19 38 ” 1” 205 k 6 1.85 k .I0 5.5 + .6 73 2 4

TP 19 35 + I** 206 2 2 0.90 + .05* 0.1 + .I* 35 2 5***
TP + Cl 16 35 r 1** 211 24 1.01 2 .15* 1.2 + .7** 52 ” 7**

(LMean k SEM.
* Different from control by Newman-Keuls test: P < 0.01.
** Different from control by Newman-Keuls test: P < 0.05.
*** TP vs TP + Cl by Newman-Keuls: P < 0.05.
258 McEWEN ET AL.

ovarian function. The behavioral effects of C1628 are consistent with the
hypothesis that T exerts some of its effects on brain sexual differentiation
after being converted to estradiol. Further support for this notion will be
offered in Experiment 3.
The failure of C1628 to block TP-induced interruption of ovarian func-
tion stands in contrast to other reports that another estrogen antagonist,
MER25, will attenuate TP-induced anovulatory sterility, as well as
anovulatory sterility induced by a synthetic estrogen, RU2858 (Doughty
and McDonald, 1974: Doughty et al., 1975b). The most likely explanation
for the difference is that neonatal exposure of female rats to MER25 does
not disrupt ovarian function, whereas C1628 does have a detectable effect
(see Experiment 3). It should also be noted that MER25 is not readily
soluble, even in sesame oil, and that it has proven to be somewhat
unreliable in the hands of various investigators in neonatal treatment
experiments. Several published studies with MER25 failed to confirm its
ability to block TP-induced anovulatory sterility (Brown-Grant, 1974;
Hayashi, 1974) or to prevent neonatal TP treatment from suppressing
lordosis behavior in hamsters (Gottlieb, Gerall, and Thiel, 1974).
EXPERIMENT 3
Experiment 2 did not take into consideration possible disruptive effects
on sexual differentiation which (31628itself might have produced. And, in
view of the results of Experiment 2 concerning the antagonistic action of
CT628on defeminization of the LQ by TP, it was of interest whether C1628
might block defeminizing effects of endogenous T in newborn male rats.
Experiment 3 was designed to study these two issues.
Procedure
Timed pregnant females (CD strain, Charles River) delivered litters of
pups in our animal facility. On Day 2 (day of birth = Day l), males and
females were given 50 ~1 of water or 200 pugof C1628 in 50 ~1 of water by
subcutaneous injection in the back. Collodion was used to seal the injec-
tion wound against leakage, and pups were placed with the mother. The
animals were weaned at 21-22 days of age. The age of vaginal opening
was recorded and is reported in Table 3. Ovariectomies were performed
when females were 76-80 days of age and orchidectomies at 88-93 days
old. Testing for lordosis was done when rats were 118-143 days of age.
Results
Females treated with 200 pg of C1628 on Day 2 showed a significantly
earlier age of vaginal opening, and on Days 76-80 their ovaries showed
significantly fewer corpora lutea than controls (Table 3). Lordosis quo;
tients under EB+P replacement therapy were also significantly reduced
by neonatal C1628 treatment (Table 3). Males treated with 200 Fg of C1628
 TABLE 3
Effect of Neonatal Treatment with CI628 on Sexual Differentiation in
Male and Female Rats

Treatment Number Day of Body wt Ovarian wti Corpora LQ

of rats vaginal (Days 76-80) body wt lutea/
opening (mdg) section
Female
: Control 23 39 f 1 261 k 4 1.47 2 .06 6.4 2 .5 81 f 3
CI628 19 35 t 1* 276 5 7 1.32 ‘- .06 4.0 2 .6** 66 2 5***
Male
Control 24 347 k 10 - 26 ” 5
C1628 34 - 346 -r- 7 - - 53 + 5*

Mean + SEM.
* P < 0.001, Students t test.
** P < 0.01, Students t test.
***P < 0.02, Students r test.
260 McEWEN ETAL.

on Day 2 showed a significantly higher LQ under EB+P replacement

therapy than controls (Table 3). The mean LQ score for the CI628-treated
males is essentially the same as that of the female rats treated neonatally
with CT628 (Table 3).
Discussion
The major finding of Experiments 2 and 3 is that neonatally adminis-
tered C1628 attenuates the defeminizing effects on the LQ of exogenous
TP in female rats and of endogenous T in male rats. That it may do this by
interfering with the action of estradiol derived from neural aromatization
of T is supported by another study from our laboratory (Lieberburg et
al., 1977) that cell nuclear levels of 3H-labeled estradiol, derived from an
injection of [3H]T into 5-day old female rats, were reduced by prior
administration of C1628 on Day 3. Tissue levels of [3H]E, were unaltered
by CI628, suggesting that aromatization was unaffected. Likewise unal-
tered were tissue and cell nuclear levels of [3H]T and [3H]DHT.
Whereas the ability of CI628 to attenuate defeminizing effects of TP in
females can be interpreted unambiguously as an interference with steroid
action, the effect of C1628 in males could have been due to a suppression
of endogenous T secretion. This seems unlikely, however, in view of the
failure of CT628to suppress basal LH secretion in ovariectomized female
rats (Redmond, Krey, unpublished).
C1628 is weakly estrogenic with respect to promotion of uterine growth
and pituitary glucose-6-phosphate dehydrogenase activity (Luine and
McEwen, 1977). It is also estrogenic with respect to the induction of
choline acetyltransferase and the suppression of type A monoamine
oxidase activity in the adult female rat brain (Luine and McEwen, 1977).
Therefore, it is understandable that CI628 did exert weak estrogenic
effects in the present experiment, mimicking weakly the defeminizing
action on ovarian function and lordosis responding which are produced
more profoundly by estradiol and by diethylstilbestrol and 1l/3-methoxy-
17a-ethynyl-17/Sestradiol (RU2858), as well as by T (Kincl, Pi, Maqueo,
Lasso, Oriol, and Dorfman, 1965; Ladosky, 1967; Clemens, 1974;
Doughty et al., 1975a). And, as already noted in the Discussion of Exper-
iment 2, it is perhaps because of these weak estrogenic effects that C1628
failed to attenuate the disruption of ovarian function produced by
neonatal TP. On the other hand, for reasons which remain to be eluci-
dated, CI628 did attenuate the defeminizing effects of neonatal TP on the
LQ despite its ability to defeminize this behavioral endpoint to a slight
degree.
GENERAL DISCUSSION
The major findings of the studies reported in this paper are that neonatal
treatment of rats with an estrogen antagonist (CI628) and an inhibitor of
 BRAIN SEXUAL DIFFERENTIATION 261

the conversion of T to E, (ATD) interfere with certain of the defeminizing

effects of exogenous neonatally administered T or TP in female rats and
of endogenous T in the male rat. These findings and those of other
laboratories (see Discussions of Experiments l-3), taken together with
biochemical evidence that C1628 and ATD selectively affect the estrogen
pathway of androgen action (Lieberburg et al., 1977), strongly support a
role for aromatization in the sexual differentiation of the rat brain.
There is general agreement that both T and E2, as well as several
synthetic estrogens, reproduce defeminizing effects on ovarian function
and lordosis behavior when given neonatally to female rats or to neona-
tally castrated male rats (see introduction). There are also many indica-
tions that nonaromatizable androgens such as DHT are ineffective in this
regard (McDonald and Doughty, 1972: Whalen and Rezek, 1974: Koren-
brot et al., 1975). However, there are also several recent papers
which indicate that DHT is capable of exerting some defeminizing effects
on development of the capability to show lordosis in the rat and hamster
(Gerall, McMurray, and Farrell, 1975: Gerall, Dunlap, and Wagner, 1976;
Payne, 1976). Whereas these effects of DHT do not appear to be as
profound as those of T or E,, they are indications that both aromatization
and 5a reduction of T may be involved in rat and hamster brain sexual
differentiation. This appears to be the case in the guinea pig, where
aromatizable androgens and not DHT are able to defeminize lordosis
behavior, while DHT and other nonaromatizable androgens appear to
have a role in the development of masculine copulatory behavior
(Goldfoot and van der Werf Ten Bosch, 1975). Further studies must
therefore assess the relative contributions of aromatization and 5a reduc-
tion to sexual differentiation in the rat and hamster brains and must bear
in mind that defeminization of feminine characteristics and masculiniza-
tion of male characteristics are separable processes which may be under
separate hormonal control (Coy and Goldfoot, 1975).
ACKNOWLEDGMENTS
Supported by Research Grant NS 07080 from the USPHS and by institutional Grant RF
70095 from The Rockefeller Foundation. We wish to thank Ms. Freddi Berg for editorial
assistance.
We gratefully acknowledge the advice and support of Dr. Donald Pfaff and members of his
laboratory in the conduct of tests for lordosis behavior.

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