Sexual differentiation of odor and partner preference in the rat.

Previous studies have shown that adult male rats, in which brain estrogen formation was inhibited neonatally by SC administration of the aromatase inhibitor 1,4, 6-androstatriene-3,17-dione (ATD), show an altered sexual partner preference. When tested in a three-compartment box, such gonadally intact ATD males approach and mate both with the estrous female and the sexually active male, whereas normal males prefer to approach and mate with the estrous female, avoiding the stimulus male. After castration in adulthood and estradiol treatment, ATD males prefer sexually active males. Similarly treated normal males prefer estrous females, and estrous females prefer to mate with males. In the present study, we asked what stimulus characteristics of active males vs. estrous females determined the different sexual preferences of males, ATD males, and of females. Were they chemosensory cues or more distal cues such as actually seeing and hearing the stimulus animals or the reward of sexual activity with the stimulus animals? Sex differences in preference were evident when animals were given a choice between soiled bedding from estrous females and from sexually active males. ATD and control males spent significantly more time on soiled bedding from estrous females than on soiled bedding from sexually active males. Control females spent significantly more time on soiled bedding from sexually active males than on soiled bedding from estrous females. More distal cues, such as seeing and hearing the stimulus animals, revealed differences in preference between control males and females, but not between ATD and control males. Physical interaction with the stimulus animals was a prerequisite for revealing differences in preference between ATD and control males. Then, the behavior of ATD males was clearly intermediate between that of normal male and female rats. In conclusion, neonatal estradiol is important for the psychosexual development of the male rat. However, the present data suggest that the psychosexual development of the male rat also requires either prenatal estradiol or perinatal testosterone.

Studies on the role of TRH and corticosterone in the regulation of prolactin and thyrotrophin secretion during lactation.

This study describes the effects of litter size and acute suckling on the synthesis and release of hypothalamic TRH, as indirectly estimated by determination of hypothalamic prothyrotrophin-releasing hormone (proTRH) mRNA and median eminence TRH content. The effects of litter size (five or ten pups) were studied throughout lactation, while suckling-induced acute changes were analyzed on day 13 of lactation in dams with ten pups. In view of the enhanced adrenal activity during lactation and recent evidence that corticosteroids have negative effects on hypothalamic TRH, we also studied adrenalectomized (ADX) dams treated with corticosterone to maintain basal plasma corticosterone levels. In addition to an increased plasma level of prolactin (PRL), adrenal weight and plasma corticosterone increased, while plasma TSH, tri-iodothyronine (T3), thyroxine (T4) and free T4 (FT4) levels decreased during lactation. Litter size correlated positively with plasma PRL, adrenal weight and plasma corticosterone. No effect of litter size was observed on plasma T3, but rats with ten pups had lower plasma TSH, T4 and FT4 than rats with a five-pup litter. Compared with dioestrous rats, lactating rats showed an increased hypothalamic proTRH mRNA content on day 2, but not on days 8 and 15 of lactation. Median eminence TRH in lactating rats gradually increased until day 15 and decreased thereafter. Acute suckling, after a 6-h separation of mother and pups, rapidly increased plasma PRL and corticosterone in the mothers, but had no effects on plasma TSH and thyroid hormone levels. Hypothalamic proTRH mRNA increased twofold after 0.5 h of suckling, and then gradually returned to presuckling values after 6 h. Compared with sham-operated rats, corticosterone-substituted ADX rats with ten pups had increased plasma PRL and TSH, hypothalamic proTRH mRNA and pituitary TSH beta mRNA on day 15 of lactation. Moreover, while acute suckling did not enhance TSH release in sham-operated rats, it provoked not only PRL but also TSH release in corticosterone-substituted ADX dams. It is concluded that suckling exerts a rapid, positive effect on hypothalamic proTRH mRNA content. However, the concurrent enhanced adrenal activity has negative effects on hypothalamic proTRH gene expression resulting in a suppressed hypophysial-thyroid axis during lactation. While TRH appears to play a role in PRL release during the first days of lactation and during acute suckling, TRH seems not important in maintaining PRL secretion during continued suckling.

Endogenous reproductive hormones and nocturnal rhythms in partner preference and sexual behavior of ATD-treated male rats.

Male rats received subcutaneously silastic capsules, containing the aromatase inhibitor 1,4,6-androstatriene-3,17-dione (ATD), shortly after birth. Control males were given silastic capsules containing cholesterol. The capsules were removed at the age of 21 days. In adulthood, blood serum was collected early and late in the dark phase of the light/dark cycle (experiment I). Testosterone and luteinizing hormone and follicle stimulating hormone (FSH) fluctuated nocturnally, both in ATD and control males, with highest levels late in the dark phase. FSH levels were significantly higher in ATD males. Nocturnal levels of inhibin, a selective suppressor of pituitary FSH secretion, also fluctuated in both ATD and control males, with lowest levels late in the dark phase. In experiment II, ATD and control males were tested for partner preference behavior in a three-compartment box (choice: sexually active male vs. estrous female) early and late in the dark phase. When gonadally intact, ATD males, but not controls, showed a clear nocturnal rhythmicity in partner preference behavior and sexual behavior. Early in the dark phase, such ATD males preferred the vicinity of and interaction with a sexually active male. Late in the dark phase, this preference for the active male shifted to a preference for the estrous female. Control males preferred the estrous female. After castration and subsequent treatment with testosterone via silastic capsules, which ensured constant blood serum levels, ATD males continued to show their nocturnal rhythms in partner preference behavior and in sexual behavior. Thus, the underlying mechanism of the nocturnal rhythmicity phenomenon is an organizational effect of neonatal ATD treatment rather than an activational effect of fluctuating serum hormone levels.

Postweaning housing conditions and partner preference and sexual behavior of neonatally ATD-treated male rats.

Male rats were neonatally treated with cholesterol or a substance that blocks the aromatization of testosterone to estradiol (1,4,6-androstatriene-3,17-dione: ATD). At weaning (21 days) they were either housed alone or in small groups (2-3 animals) and tested for partner preference behavior (PPB) in adulthood. Choice was between an estrous female and an active male (Part I) and between an estrous female and an ATD-male (Part II). Tests were carried out in a 3-compartment box. Social isolation did not have major effects on PPB except when sexual interaction with the stimulus animals was prevented (Part I). In this case, isolates (ATD and control) showed higher preference scores (PS) for the estrous female and spent less time in the empty middle compartment. When the choice was between an estrous female and an ATD-male, partner PS decreased in all males, most clearly in ATD-males. The latter animals spent more time with the stimulus ATD-male than they had done in previous PPB tests with the normal stimulus male. In contrast to partner preference behaviors, sexual behavior was clearly affected by social isolation. Isolates (ATD and control) displayed lower frequencies of mounts and intromissions. These effects persisted over consecutive tests. Ejaculation was not affected. In conclusion, the present results confirm earlier findings about the significance of neonatal endocrine conditions for the organization of adult PPB in male rats. The presence or absence of social conspecifics after weaning appears to have little influence on adult PPB.

A semiautomated test apparatus for studying partner preference behavior in the rat.

A semiautomated three-compartment box (3CB) for studying partner preference behavior of rats is described. This apparatus automatically records the rat's time spent in each compartment, as well as the locomotor activity (i.e., the number of visits an animal pays to each compartment). Software was developed for calculating partner preference scores. Behavioral testing in the semiautomated 3CB, which is a modification of an earlier version, is less time consuming and less laborious. Three 3CBs can be observed simultaneously by two trained observers, and the behavioral interactions of three experimental animals with the stimulus animals can be observed and scored by hand. The use of the new apparatus was validated by studying adult partner preference behavior of neonatally ATD-treated male rats. The collected data fully corroborate previous results, obtained in the earlier version of the 3CB, again revealing the behavioral bisexual nature of these ATD males. A new finding was the much higher locomotor activity of the ATD males compared to controls.

Organization of partner preference and sexual behavior and its nocturnal rhythmicity in male rats.

Male rats were neonatally treated with 1,4,6-androstatriene-3,17-dione (ATD), which blocks aromatization of testosterone (T) to estradiol (E2), from Days 0, 2, or 5 through 14. Adult partner preference behavior (PPB; choice between estrous female rat [F] and active male rat [M]) was studied in the early part of the dark phase of the light-dark (LD) cycle. ATD Day 0 Ms showed a preference for the stimulus M or showed no preference for either of the stimulus Ss. Controls preferred the estrous F. ATD Days 2 and 5 Ms showed PPB intermediate between ATD Day 0 Ms and controls. Thus the neonatally sensitive period for organization of adult PPB extends beyond Day 5. Furthermore, PPB showed a nocturnal rhythmicity in ATD Ms but not in controls. In the late part of the dark phase, all Ms showed a preference for the stimulus F. ATD Days 2 and 5 Ms and control Ms were no longer different in PPB, but ATD Day 0 Ms still showed significantly lower preference scores for F than all other Ms. Thus the E2 metabolite of T suppresses organization of an adult nocturnal rhythm in PPB.

Hormonal regulation of adult partner preference behavior in neonatally ATD-treated male rats.

Male rats, neonatally treated with ATD (1,4,6-androstatriene-3,17-dione), which blocks the aromatization of testosterone into estradiol (E2), were tested for adult partner preference behavior (PPB; estrous female vs. active male). Castration caused a decrease in preference for the female partner in all males, with ATD males showing lower preference for the female partner than controls. Long-term castrated males did not show preference for either partner. Precastration levels of PPB in control males occurred after treatment with E2 or dihydrotestosterone (DHT) plus E2. DHT alone had no effect on PPB. With E2 alone, the ATD males clearly preferred the male partner. When DHT was added, these ATD males showed no preference for either partner or a low preference for the female partner. In conclusion, adult PPB in male rats is activated by endogenous testosterone or by both its metabolites (DHT and E2) or by E2 alone. ATD males showed a much lower preference for the female. There was a differential effect of DHT and E2: DHT had no effect, but E2 clearly caused ATD males to prefer the male partner and control males to prefer the female partner.

Copulatory stimuli in rats induce heat abbreviation through effects on genitalia but not through effects on central nervous mechanisms supporting the steroid hormone-induced sexual responsiveness.

Male copulatory stimuli in various animal species abbreviate the length of the period of their female conspecifics' heat. Such effect can be explained in some species as the result of copulation-induced alterations in ovarian hormone secretion (e.g. reflex ovulators). Other species require a different explanation: specifically, interactions in the hypothalamus between effects of steroid hormones and inhibitory neural afferents from the genital area have been hypothesized to play a role in some laboratory animals such as guinea pigs, hamsters and rats. However, the evidence in support of heat abbreviation through copulatory stimuli in rats is at best equivocal. There is obvious need for its unequivocal demonstration prior to any further analysis of the potential mechanism of action. The present study intended to find a model in support of the concept of 'centrally induced' heat abbreviation in rats. Virgin rats during 'natural heat' were examined with or without allowing them the opportunity to pace male copulatory activities. Ovariectomized, steroid hormone-treated rats were studied with minor or ample experience with sexual activities in prior tests and, also, with or without the opportunity to 'pace' male sexual acts. None of the experimental models revealed unequivocal support for detrimental effect of large numbers of intromittive copulations on 'central mechanisms' regulating heat duration. It rather seemed that frequent intromittive copulations and ejaculates affected the 'peripheral' genital tract (vulva, vagina, cervix) making further copulation of a highly aversive quality so that sexual encounters were avoided or prevented. This effect was noticed with great inter-individual variability. The conclusion is drawn that rats do not show the counterpart of heat abbreviation reportedly occurring rapidly and reliably in guinea pigs after a limited amount of vaginal/cervical stimulation through copulation or insertions of a glass-rod.