Effects of neonatal septal lesions on reproductive behavior of male and female rats

The purpose of this study was to examine the effects of neonatally placed septal lesions (SL) in male, female, and androgenized female rats on reproductive behavior. Animals were castrated as adults and tested for both feminine and masculine sexual behavior. After treatment with estradiol benzoate (EB) alone (2 μg daily for 3 days), only the females with SL which had not been given testosterone propionate (TP) neonatally showed a facilitation of lordosis behavior. Following EB (2 μg for 3 days) plus 0.5 mg progesterone (P), both the lesioned and the sham-operated female groups showed an increase in the display of lordosis in either hormonal condition. All animals were given a pretest for masculine sexual behavior and tested on Days 4, 7, 11, and 15 of daily TP treatment (150 μg/day). There was no effect of the neonatally placed SL on masculine sexual behavior in female rats or in female rats androgenized with 30 μg TP. However, lesioned females treated neonatally with 1 mg TP showed a marginal enhancement of masculine sexual behavior. Male rats given SL neonatally showed a marked enhancement of masculine sexual behavior compared to that of controls. These results suggest that, depending on the neonatal hormone environment, SL selectively increase behavioral sensitivity to hormones. Although neonatally lesioned females show behavioral responses similar to females given SL as adults, male rats given SL neonatally are unique in that they show enhanced masculine sexual behavior whereas males lesioned as adults do not.     

Neonatal androgen and sexual behavior in female house mice

A series of experiments investigated masculine response potential in normal BDF₁ female mice and in females who had been injected with 100 μg of testosterone propionate on the day of birth. Sixty-five percent of BDF₁ females mounted females in estrus; ovariectomy lowered this response potential while injections of TP in adulthood raised masculine RP in both ovariectomized and intact females. Neonatally androgenized females with or without TP in adulthood exhibited the full range of masculine responses including the ejaculatory reflex. Comparisons of elements of sexual behavior are made between neonatally androgenized females and normal males.     

Sexual differentiation of the steroid feedback mechanism regulating follicle-stimulating hormone secretion in the Syrian hamster

The ability of gonadal steroid hormones to influence tonic follicle-stimulating hormone (FSH) secretion was investigated in Syrian hamsters. In Experiment 1, males were castrated as adults, and administered testosterone in 20-, 30-, 40-, and 50-mm silastic capsules (s.c.) at 67, 74, 81, and 88 days, respectively. Circulating FSH was reduced by testosterone in a dose-dependent manner. A similar FSH response to testosterone in adulthood was evident in neonatally androgenized hamsters given testosterone proprionate (TP) on Days 0 and 1 of life. By contrast, the absence of gonadal androgens during the neonatal period (females ovariectomized at 60 days of age and males orchidectomized at birth) resulted in only a partial suppression of circulating FSH by even the highest dose of testosterone during adulthood. Treatment with estradiol benzoate at birth failed to produce a masculine response to androgen in adulthood. In Experiment 2, using a similar protocol, the nonaromatizable androgen, dihydrotestosterone, produced a dose-dependent suppression in serum FSH in males castrated in adulthood (30-, 60-, 90-mm capsules). However, dihydrotestosterone failed to alter the hypersecretion of FSH produced by orchidectomy at birth in males or in females ovariectomized at 60 days of age and treated neonatally with either vehicle or TP. In Experiment 3, treatment with estradiol (10-, 20-, 30-mm capsules) decreased serum FSH in gonadectomized hamsters in a dose-dependent manner; males and females treated neonatally with TP were more responsive to estradiol as adults compared to neonatally orchidectomized males or females treated with vehicle at birth.(ABSTRACT TRUNCATED AT 250 WORDS)     

Attractivity of male and female rats which are hormonally manipulated during early development and in adulthood

Attractivity is one aspect of female sexuality relevant for the understanding of male-female sexual interactions. In a previous study, it was shown that intact males were equally attracted to early androgenized, gonadally intact females as to normally developed, estrous females. The present study was designed to investigate in what way hormones given in adulthood might influence attractivity of early androgenized females in adulthood. Specifically, we compared the attractivity of neonatally androgenized females (NeoTP) to the attractivity of normally developed females (NeoOIL), neonatally castrated males (NeoCASTR), and neonatally sham-castrated males (NeoSHAM) when different groups received either OIL, estradiol benzoate (EB) or testosterone propionate (TP) in adulthood. The male's preference to stay in the vicinity of one incentive in favor of the other was taken as an index of attractivity. The results show that, under the present hormonal conditions, NeoTP-females are generally less attractive than NeoOIL-females, more attractive than NeoSHAM-males, and equally attractive as NeoCASTR-males. TP-treated androgenized females were found to be equally attractive as TP-treated NeoSHAM-males. It is concluded that, relative to normally developed females, androgenized females become less attractive when the endogenous secretion of sex steroids is artifically controlled by gonadectomy and/or by administration of fixed amounts of sex steroids.     

Influence of androgen in the neonatal period on ejaculatory and postejaculatory behavior in the rat

The objective of the present report was to investigate the influence of androgen in the neonatal period on the development of ejaculatory and postejaculatory behavior. At birth, male rats were either castrated (neonatally castrated males), implanted with a Silastic tube of the aromatase inhibitor androsta-1,4,6-triene-3,17-dione for the first 10 days (ATD males), or left untreated (normal males). Female rats were either injected with 0.5 mg testosterone propionate (TP) on Days 1 (day of birth) and 2 (androgenized females) or left untreated (normal females). All gonadally intact animals were castrated at 60 days of age. Following TP administration, all animals were tested for ejaculatory and postejaculatory behavior under both shock and nonshock conditions. All animals were capable of showing the intromission pattern; however, the ejaculatory pattern was exhibited regularly only by those animals exposed to androgen at birth (normal males, androgenized females, and ATD males). The normal males required fewer intromissions to achieve ejaculation than the other two groups exhibiting this reflex. This result is discussed in terms of peripheral genital stimulation deficits and the differentiation of neural tissue responsible for masculine copulatory behavior. Androgenized females and ATD males displayed a refractory period, characterized by 22-kHz vocalizations, equal to or longer than that found in normal males. These results indicate that defeminization is not necessary for the display of normal ejaculatory and postejaculatory behavior.     

Masculine sexual behavior in male and female rats following perinatal manipulation of androgen: Effects of genital anesthetization and sexual experience

Androgenized females, Day 1 male castrates, normal males, and normal females were tested for mounting behavior as adults following TP administration (1 mg/day). Genital anesthetization was used to eliminate all intromission and ejaculation behavior. Results showed that sexually-naive normal males and androgenized females mounted significantly more then Day 1 male castrates, while the Day 1 male castrates mounted significantly more than normal females. Sexually-experienced normal males and androgenized females displayed a significant facilitation of mounting behavior as compared to the sexually-naive animals. Tests of masculine copulatory behavior without genital anesthetization also were conducted with androgenized females and normal males. In these tests, androgenized females were indistinguishable from normal males in all aspects of the complete masculine pattern. The present results provide evidence that during perinatal development androgen acts directly on neural systems which will later regulate adult masculine sexual behavior.     

Influence of fetal position on neonatal androgen-induced sterility and sexual behavior in female rats

Two aspects of reproductive function were examined in relation to female fetus' contiguity to intrauterine male littermates. Following injection of 3.5 μg testosterone propionate (TP) on Day 3, females that had been positioned in utero between two males became sterile earlier in life than those located in utero between two females. Anogenital distances on Days 1 and 3 prior to neonatal treatment with TP were greater in females located in utero between two males than in females residing between two females in utero, suggesting that females developing between two males may have been exposed prenatally to a masculinizing substance, presumably an androgen. No reliable contribution of litter composition was apparent with respect to differentiation of female sexual behavior. Results indicate that littermate hormonal influence is present or effective and can be detected in a neonatally androgenized preparation.     

Influence of androgen on the development of sexual behavior in rats : I. Time of administration and masculine copulatory responses, penile reflexes, and androgen receptors in females

The objective of the present study was to investigate the effect of the time of administration of androgen, during the neonatal period, on the development of masculine copulatory behavior in female rats. In addition, the influence of androgen, administered neonatally, on the development of penile reflexes and cytoplasmic androgen receptor levels in the hypothalamic-preoptic area (HPOA) was examined. Female rats were injected with 0.5 mg testosterone propionate (TP) at either 1, 8, or 24 hr after birth and again 24 hr after the first injection. Fifty percent of the females treated with TP at 1 and 8 hr after birth displayed the ejaculatory response when tested in adulthood. In contrast, 93 and 87.5% of oil-treated males and females, respectively, which were androgenized at 24 hr after birth exhibited this response. The results indicate that a considerable amount of masculinization occurs postnatally in the rat. However, none of the androgenized females displayed any penile reflexes even when tested following the display of an ejaculatory response. HPOA androgen receptor levels were somewhat higher in the oil-treated females than in males but were not correlated with the ability to exhibit ejaculation patterns.     

The sexual behaviour of prenatally androgenized ewes observed in the field

Pregnant ewes were implanted with 1 g testosterone between Days 30-80, 50-100, 70-120 or 90-140 of gestation. Treatments which began on Days 30, 50 or 70 resulted in the birth of androgenized females which failed to show regular oestrous cycles in adult life, but which exhibited patterns of male-like behaviour. This was most marked in the Day 50-100 and 70-120 groups, whereas complete masculinization of the external genitalia was confined to the Day 30-80 group. Animals in the Day 90-140 group had regular oestrous cycles although they showed slight enhancement of masculine behaviour compared to the control ewes. These results demonstrate that androgenization involves both a suppression of female behavioural patterns, and the development of male patterns; these are not mutually exclusive.     

Behavioral masculinization is independent of genital masculinization in prenatally androgenized female rhesus macaques

Genetic female fetuses were exposed transplacentally to testosterone propionate injected into their mothers either early (Days 40 through 64) or late (Days 115 through 139) in gestation. Early and late androgenized females (EAFs and LAFs, respectively) were raised with normal males and females that served as criteria for evaluating degree of behavioral masculinization induced by the prenatal androgen. EAFs were genitally virilized and LAFs were not. Males and untreated females differed reliably on five behavioral measures: males showed more mother-mounting, more peer-mounting, more rough play with peers, a preference for initiating play with male partners, and less grooming of mothers. Neither type of prenatally androgenized female showed masculinization of all five types of behavior. Compared with females, EAFs showed more mother-mounting, more peer-mounting, less mother-grooming, did not differ from females in rough play, and did not manifest a preference for male partners. LAFs, like females, groomed but did not mount their mothers, and did not show a preference for male partners; but unlike females they showed elevated rough play and mounting with peers. EAFs showed a statistically significant delay in puberty onset (menarche), but LAFs did not. Mothers inspected genitalia of their offspring more often if they were males than if they were females. Mothers of EAFs inspected their offspring's genitalia as often as mothers of males, but mothers of LAFs did not. No aspect of maternal behavior was associated with either the amount or kind of masculine behavior shown toward peers. We interpret the results to mean that genital virilization is independent of, and largely irrelevant to, the expression of those behavioral traits that characterize the juvenile male social role. Moreover, the individual behavior traits that are components of the juvenile male role are independently regulated by the organizing actions of androgen and have separable critical periods. Of the two major traits, mounting peers and rough play with peers, the latter has a greater requirement for androgenic stimulation late in prenatal life.     

Role of postnatal androgens in sexual differentiation of the lordosis-inhibiting effect of central injections of cholecystokinin

The neuropeptide cholecystokinin (CCK) inhibits lordosis behavior when infused into the ventromedial nucleus of the hypothalamus (VMN) of female rats and has no effect when infused into the VMN of male rats. To test whether this sex difference develops under the control of perinatal steroids, male rats were castrated or given sham surgeries within 3 h of birth and female rats were injected with either 0 or 100 micrograms testosterone propionate on postnatal day 5. As adults, these rats were castrated as necessary, implanted with unilateral cannulae directed at the VMN, and tested for their ability to display female sexual behavior and to respond to CCK. Neonatal castration of males prevented defeminization of this response. When treated with 5 micrograms estradiol benzoate (EB), neonatally castrated males showed both lordosis behavior and a profound inhibition of that behavior after infusions of CCK. Neonatally castrated males did not display lordosis behavior when treated with 2 micrograms EB. Control males showed no lordosis behavior and, therefore, no response to CCK. Both doses of EB induced lordosis behavior in neonatally androgenized females. Significantly, these neonatally androgenized females were less responsive to CCK's inhibition of lordosis and were also anovulatory. These results imply that androgens alter the development of CCK responsive circuits as well as defeminize cyclic gonadotropin release. Levels of 125I-sCCK-8 binding in the VMN were correlated closely with an individual's ability to respond to sCCK-8. In summary, the inhibition of female sexual behavior caused by exogenously administered CCK in normal adult female rats appears to be controlled at least partially by levels of CCK receptors in the VMN and to differentiate under the control of perinatally present testosterone.     

Period of maximal susceptibility to behavioral modification by testosterone in the golden hamster

Male hamsters castrated on the day of birth (Day 1) and female hamsters were treated with the free form of testosterone (100 μg/day) on Days 1 and 2, 3 and 4, 5 and 6, 7 and 8, or 9 and 10 postnatally. Following androgen treatment in adulthood, animals treated on Days 1 and 2 or 3 and 4 showed significantly higher mounting and intromission frequencies than animals treated later in life. Sexual receptivity measures following ovarian hormone treatment showed no differences among the male groups, whereas females treated on Days 1 and 2 or 3 and 4 were significantly lower in sexual receptivity measures than females in other treatment groups. Histology of the adult ovaries indicated no modification of normal function in any treatment group. In a subsequent experiment, Day 1 castrated male and intact female hamsters were treated with the free form of testosterone on Days 1–5 (40 or 100 μg/day), 6–10 (40 or 100 μg/day), or Days 1–10 (50 μg/day). Masculine behavior measures were significantly higher in males treated Days 1–10 than in other groups. Among the females, masculine behavior was highest in those treated Days 1–5 postnatally. Sexual receptivity in both males and females was significantly depressed by testosterone treatment Days 1–10 postnatally. Ovarian histology also revealed alterations in gonadal function in females treated Days 1–5 and 1–10 postnatally. Compared with previously published findings, these data suggest that testosterone can be as effective in inducing behavioral masculinization and defeminization as testosterone propionate, provided that treatment extends over a prolonged period during early postnatal development.     

Regulation of urine marking in male and female mice: effects of sex steroids

Effects of sex steroids on urine-marking activity were studied in male, female, and neonatally androgenized female mice. Urine marking was estimated by suspending ceramic tubes that were connected in a horizontal row with a steel rod into the home cage of an isolated mouse. Intact males showed high marking activity, which was diminished after castration. Both testosterone propionate (TP) and estradiol benzoate (EB) were effective in restoring the marking activity of castrated males, while 5-alpha-dihydrotesterone (DHT) did not have any stimulative effects. Intact normal females showed quite low marking activity and ovariectomy further depressed it. TP and DHT enhanced the marking of ovariectomized females, but EB restored the activity only to the preovariectomy level. In intact females which were neonatally androgenized, the marking activity was much higher than that of normal females. The pattern of the change induced by gonadectomy and hormone treatment in these females resembled that in males. Thus, ovariectomy reduced the activity and both TP and EB restored the level. These results indicate that the sexual dimorphism in the urine marking in mice is primarily determined by hormonal environment during early postnatal age. Hormonal control of scent marking is discussed in relation to the studies in other rodents.     

Reversal of reproductive deficiency in the hpg male mouse by neonatal androgenization.

Some aspects of reproductive function in the GnRH-deficient hypogonadal (hpg) mutant mouse can be restored by transplanting normal fetal brain tissue containing GnRH cells into the central nervous system of adult hpg mice. However, hpg males showing physiological response to the graft fail to display sexual behavior and are infertile. We hypothesized that the reproductive deficit of these males is due to insufficient perinatal exposure to testicular androgens as a consequence of the GnRH deficiency. To test this hypothesis we androgenized hpg males by giving them neonatal injections of testosterone propionate (TP). Controls consisted of hpg males not androgenized neonatally and of normal males. All three groups received a TP implant in adulthood, and their copulatory behavior and reproductive capability were recorded. In addition, other hpg males, not androgenized neonatally, received fetal brain transplants containing GnRH neurons and were also tested for copulatory behavior and reproductive capability before and after receiving a TP implant. Three of 8 neonatally androgenized hpg males expressed the full repertoire of male sexual behavior, including intromission and ejaculation, and sired several litters. Three of 7 control hpg males that were not androgenized neonatally but received TP implants in adulthood also displayed mounting and intromission, but there was no evidence of ejaculation, and these males failed to impregnate normal females. Of the 8 hpg males that responded to a fetal transplant with testicular growth, only 1 displayed mounting behavior. However, when given a TP implant, 4 of 8 hpg males with grafts displayed mounting and intromissions.(ABSTRACT TRUNCATED AT 250 WORDS)     

Copulatory behavior of adult tamarins (Saguinus fuscicollis) castrated as neonates or juveniles: Effect of testosterone treatment

The sexual interactions of Saguinus fuscicollis males castrated as neonates, at 37 days of age, or prepubertally with adult intact females were studied. Prepubertally castrated males were observed while receiving testosterone, and while being treated with saline. Males castrated neonatally or at 37 days of age were observed while receiving testosterone. Neonatal castrates had previously been studied without hormone treatment and therefore no control condition was included for these animals. Prepubertally castrated males showed Mounts, Mounts with Thrusts, and Sexual Tongue Flicking when treated with saline only. In three of the four males, all measures of sexual behavior increased with testosterone treatment. Neonatally castrated males had failed to display any mounting or thrusting without testosterone treatment during a previous study. During the present study, three of the four males did not respond to testosterone treatment with sexual behavior. The fourth male and one male castrated at 37 days of age displayed some sexual behavior. These results suggest that most neonatally castrated males are not able to respond to testosterone with the activation of copulatory behavior. The findings are consistent with the hypothesis that in callitrichids the sensitive period for behavioral differentiation is shifted into neonatal life. However, some neonatally castrated males show a weak response to testosterone. This may reflect an extended and perhaps partially prenatal period of sensitivity.     

Attractivity of male and female rats after early endocrine manipulation

Approach behavior toward males and approach behavior toward females were studied in the male rat. Adult male rats approached neonatally castrated male rats more than they approached intact males. Neonatally androgenized female rats and estrous female rats were equally approached. A preferential approach toward the androgenized female was evident when the alternative incentive was a proestrous female. These adult patterns of approach behavior appeared on Day 50 after birth. It is concluded that early endocrine manipulation influences the attractivity of the incentives.     

Specific morphogenetic events in mouse external genitalia sex differentiation are responsive/dependent upon androgens and/or estrogens

The objective of this study was to perform a comprehensive morphologic analysis of developing mouse external genitalia (ExG) and to determine specific sexual differentiation features that are responsive to androgens or estrogens. To eliminate sex steroid signaling postnatally, male and female mice were gonadectomized on the day of birth, and then injected intraperitoneally every other day with DES (200ng/g), DHT (1μg/g), or oil. On day-10 postnatal male and female ExG were dissected, fixed, embedded, serially sectioned and analyzed. We identified 10 sexually dimorphic anatomical features indicative of normal penile and clitoral differentiation in intact mice. Several (but not all) penile features were impaired or abolished as a result of neonatal castration. Those penile features remaining after neonatal castration were completely abolished with attendant clitoral development in androgen receptor (AR) mutant male mice (X(Tfm)/Y and X/Y AR-null) in which AR signaling is absent both pre- and postnatally. Administration of DHT to neonatally castrated males restored development of all 10 masculine features to almost normal levels. Neonatal ovariectomy of female mice had little effect on clitoral development, whereas treatment of ovariectomized female mice with DHT induced partial masculinization of the clitoris. Administration of DES to neonatally gonadectomized male and female mice elicited a spectrum of development abnormalities. These studies demonstrate that the presence or absence of androgen prenatally specifies penile versus clitoral identity. Differentiated penile features emerge postnatally and are sensitive to and dependent upon prenatal or pre- and postnatal androgen. Emergence of differentiated clitoral features occurs postnatally in either intact or ovariectomized females. It is likely that each penile and clitoral feature has a unique time-course of hormonal dependency/sensitivity.     

Sexual orientation, proceptivity, and receptivity in the male rat as a function of neonatal hormonal manipulation

The influence of neonatal androgen on the potential to exhibit feminine sexual behavior was investigated. Male rats castrated on Day 0 but not those castrated on Day 4 or later showed hop/darting, ear wiggling, and lordotic behavior in response to treatment with estrogen and progesterone in adulthood at a frequency equal to that of females. Neonatal treatment with testosterone propionate (1 mg/rat for 4 days) abolished the capacity to show these behaviors. In subsequent experiments, involving castration of male rats at 0 or 4 hr after cesarean delivery, the effect of the postnatal surge of testicular secretions on the expression of female sexual behavior was investigated. No differences were seen in the frequency of hop/darting, ear wiggling, and receptivity between males castrated immediately or 4 hr after delivery. In a preference test where the experimental male could choose between an estrous female and a sexually active male, the neonatally castrated males preferred the company of a male when treated with estrogen and progesterone. The implantation of testosterone resulted in a preference for an estrous female. It was concluded that testicular secretions in the newborn male influence adult sexual orientation and suppress the ability to show proceptive and receptive behaviors.     

Anxiolytic effects of progesterone are sexually dimorphic

We evaluated the effects of progesterone administration to estrogen-primed rats on licking behavior during a lick suppression test. This test assesses the behavior of water-deprived rats to lick a drinking tube for water reinforcement that is paired to electric shock. Chlordiazepoxide (a commonly used tranquilizer) is very effective in increasing licking during this test. Normal females and neonatally castrated males displayed an increase in shock-punished responses in tests conducted after hormone injections. Normal males and neonatally androgenized females were unaffected. Our results suggest that ovarian steroids, and particularly progesterone, can modulate anxiety in females and feminized males. In addition, the anxiolytic effect of progesterone appears to be mediated by a mechanism different from that of Chlordiazepoxide.     

Effects of testosterone, estrogen, and dihydrotestosterone upon aggressive and sexual behavior of female rats

Groups of female TMD rats were treated either with estradiol benzoate (EB), dihydrotestosterone propionate (DHTP), testosterone propionate (TP), EB + DHTP (EB/DHTP), or with oil. These groups of females were tested for social aggression and for masculine and feminine sexual behavior. In addition, patterns of masculine and feminine sexual responses during the aggressive encounters, were investigated. TP-treated females of the same strain were used as opponents in the tests for aggression. In accordance with previous results, EB did not activate aggression whereas TP treatment resulted in a significant increase in aggression in females. Aggressive responses were activated by adding DHTP to EB, up to levels equal to those activated by TP. Sexual responses were observed in the tests for aggression as well as in tests for sexual behavior. The results indicated that feminine and masculine sexual responses were affected significantly by hormonal treatment. Mounting behavior in the test for aggression was activated by TP and by EB/DHTP. Lordosis and proceptive responses were inhibited in these groups as compared to EB-treated females, both in tests for aggression and in tests for sexual behavior. The results are consistent with the idea that dihydrotestosterone inhibits feminine and activates masculine sexual activity. The results also indicate that EB and DHTP synergistically activate aggression.