Female effect in sheep. I. The effects of sexual receptivity of females and the sexual experience of rams

When in contact with receptive or non-receptive ewes, the presence of females increased LH peak frequency both in experienced and inexperienced rams (P less than 0.05). The highest response was found in experienced rams stimulated by sexually receptive ewes. In this group only, the mean testosterone levels increased during stimulation (P less than 0.05). Sexual behavior did not differ between experienced and inexperienced males. The sexual receptivity of the females and the sexual experience of rams appear to interact and thus facilitate the rams' LH and testosterone responses to the presence of ewes.     

Role of the olfactory systems and importance of learning in the ewes' response to rams or their odors

In sheep, exposure of seasonally anestrous females to the male or its fleece results in activation of luteinizing hormone (LH) secretion and synchronized ovulation. The study of the neural pathways involved in this phenomenon, commonly named "male effect", show that the main olfactory system plays a critical role in the detection and the integration of the male odor. The accessory olfactory system participates in the perception of the ram odor but does not seem necessary for the endocrine response. According to the hypothesis that the neuroanatomical differences between the two olfactory systems could be associated with different functional roles, we investigated the importance of sexual experience and learning processes in the male effect. Our results showed that female responses depend on previous sexual experience. We also demonstrated that the LH response to male odor could result from an associative learning process. The aim of the present report was to summarize our current knowledge concerning the "male effect" and in particular to clarify the role of sexual experience and learning in the processes involved in this effect.     

Role of experience in the neuroendocrine control of ewes' sexual behavior

We assessed the role of learning in the expression of female sexual behavior and evaluated the relative importance of age versus experience. Two studies were conducted with ovariectomized ewes submitted to steroid treatment that mimicked an estrus cycle. We compared behavioral (experiments 1 and 2), neurochemical (experiment 1), and endocrine (experiment 2) responses of sexually naive young and adult ewes versus sexually experienced adults when exposed to males. In a third study, we compared their performance in an instrumental learning test and the extent to which it was affected by stress. These experiments showed that proceptivity is affected both by age and sexual experience. In experiment 1 only experienced adults were proceptive and displayed an increase in hypothalamic norepinephrine. By the second estrus cycle (experiment 2) naive adults performed similarly to experienced adults but proceptive behavior was still inferior in young ewes. Receptivity was also different between groups but affected more by age than by sexual experience. All ewes mated during the first interaction with a male, although males' latency to ejaculation was shorter with experienced females than naive adults or naive young. Young ewes found food as readily as adults in experiment 3 but were more affected by stress. Together, these experiments show that both experience and age influence sexual activity and that sensitivity to stress may also be involved. This may contribute to the deficient reproductive performance that is often observed in young female mammals.     

Lack of sexual experience does not reduce the responses of LH, estrus or fertility in anestrous goats exposed to sexually active males

We investigated whether LH secretion, estrous behavior and fertility would differ between sexually inexperienced and experienced anestrous goats exposed to the males. Male goats were rendered sexually active during the reproductive rest season by exposure to 2.5 months of artificial long days. Two groups of anovulatory sexually inexperienced and sexually experienced does were exposed to males during 15 days (n = 20 per group). LH pulsatility was determined every 15 min from 4 h before to 8 h after introducing males (Day 0). Estrous behavior was recorded twice daily. Pregnancy rates were determined on Day 50. Fertility was determined at parturition. Male sexual behavior was registered on days 1 and 2 during 1 h. Before introducing the males, the number of LH pulses did not differ between groups. After introduction of the males, all females increased their LH pulsatility, but the number of pulses did not differ between sexually inexperienced and experienced goats. The proportion of females displaying estrous behavior with a high pregnancy rate and fertility did not differ between inexperienced and experienced goats. The sexual behavior of the males did not differ significantly between those interacting with sexually inexperienced or experienced goats. We conclude that goats can show substantial endocrine and reproductive responses to males, even in the absence of previous sexual experience, when sexually active bucks are used.     

Experiential and endocrine dependence of gonadotropin responses in male mice to conspecific urine

Previous research has shown that a urinary pheromone of female mice acts via the vomeronasal organ of the accessory olfactory system to elicit rapid release of luteinizing hormone (LH) in conspecific males. Several experiments were conducted to examine the importance of sexual experience for gonadotropin responses in male mice to female urine, male urine, saline, or mixtures of these stimuli. Both sexually naive and sexually experienced male mice had significantly higher plasma LH levels after presentations of female urine than after presentations of male urine. However, sexual experience appeared to increase the reliability of the short-latency gonadotropin response to female urine relative to a sexually neutral component of urine such as sodium chloride, and male urine appeared to suppress spontaneous LH secretion episodes in both naive and sexually experienced males. Subsequent experiments with sexually experienced subjects demonstrated that male mouse urine is a powerful suppressant of LH release in other males. Specifically, female mouse urine mixed with male urine failed to elicit LH responses in male subjects, whereas female urine mixed with saline was highly effective. Urine obtained from castrated male donors was as potent as urine from intact males in suppressing the gonadotropin response to female urine. The suppressive activity in male mouse urine thus does not appear to be critically dependent on gonadal hormones. The existence of a potent stimulatory pheromone in female urine and a potent suppressive pheromone in male urine makes male mice an excellent model system for studying the neural regulation of LH secretion.     

Sexual stimulation does not affect plasma LH nor testosterone levels in New-Zealand male rabbits

Young sexually naive (3-4 months) and sexually experienced (2-3 years) male rabbits were subjected to various sexual stimulation procedures. Blood samples were taken just before and 30 min after mounting (unreceptive females) or coitus (receptive females). Testosterone and luteinizing hormone were assayed using specific radioimmunoassays. Sexual stimulation did not affect postcoital testosterone and LH levels in naive and experienced males while in females LH levels were significantly increased. We may conclude that the postcoital neuroendocrine reflex which causes a massive release of pituitary LH in female rabbit does not exist in males.     

Importance of the signals provided by the buck for the success of the male effect in goats

Under temperate and subtropical latitudes, ewes and goats display a reproductive seasonal pattern and their sexual activity during the anestrous period can be stimulated and synchronized by the introduction of males in the group, which is called the "male effect". The response of females to the male effect in the middle of the anestrous season is weak or absent. This failure may be due to the inability of the female to respond to males, as a result of a refractoriness of the female to the male stimulus. But, it may also be due to a low quality stimulus provided by the male which is, as the females, in seasonal rest. We tested this latter hypothesis in seasonally inactive goats kept under subtropical conditions by comparing the use of males with their sexual behavior stimulated or not by photoperiodic treatments. Treated males were able to induce estrous activity of females during the whole anestrus season. We have also determined that previous separation of the males and continuous contact during teasing are not absolute requisites when active bucks are used. While odor from the male and its sexual behavior play a primary role in inducing ovulation, vocalizations appear to facilitate the display of the does' estrous. It remains to be determined to which extent these conclusions apply under temperate latitudes and with more seasonal breeds.     

Lack of sexual experience does not reduce the responses of LH,estrus or fertility in anestrous goats exposed to sexually active males

We investigated whether LH secretion, estrous behavior and fertility would differ between sexually inexperienced and experienced anestrous goats exposed to the males. Male goats were rendered sexually active during the reproductive rest season by exposure to 2.5 months of artificial long days. Two groups of anovulatory sexually inexperienced and sexually experienced does were exposed to males during 15 days (n = 20 per group). LH pulsatility was determined every 15 min from 4 h before to 8 h after introducing males (Day 0). Estrous behavior was recorded twice daily. Pregnancy rates were determined on Day 50. Fertility was determined at parturition. Male sexual behavior was registered on days 1 and 2 during 1 h. Before introducing the males, the number of LH pulses did not differ between groups. After introduction of the males, all females increased their LH pulsatility, but the number of pulses did not differ between sexually inexperienced and experienced goats. The proportion of females displaying estrous behavior with a high pregnancy rate and fertility did not differ between inexperienced and experienced goats. The sexual behavior of the males did not differ significantly between those interacting with sexually inexperienced or experienced goats. We conclude that goats can show substantial endocrine and reproductive responses to males, even in the absence of previous sexual experience, when sexually active bucks are used.     

Importance of intense male sexual behavior for inducing the preovulatory LH surge and ovulation in seasonally anovulatory female goats

The present study was carried out to determine whether the presence of photostimulated sedated male goats could stimulate the LH preovulatory surge and ovulation in seasonal anestrous goats. Sexually experienced male goats were treated with artificial long days (16 hours light per day) from 1 November to 15 January to stimulate their sexual activity in March and April, corresponding to the natural sexual rest. A female group of goats (n = 20) was exposed to non-sedated males who displayed an intense sexual behavior and provided strong odor (non-sedated group). Another female group of goats (n = 20) was exposed to the photo-stimulated male goats, but these males were sedated with Xylazine 2% to prevent the expression of sexual behavior (sedated group). The sedated males also provided a strong odor. Females of both groups had full physical and visual contact with non-sedated or sedated males. In both groups, the males remained with females during 4 days. The LH preovulatory surge of 10 female goats per group was measured by determination of LH plasma concentrations in samples taken every 3 hours. In addition, in all goats, (n = 20 by group), ovulation was determined by measuring plasma concentrations of progesterone. The proportion of female goats showing a preovulatory LH surge was higher in goats exposed to non-sedated (10/10) than in those exposed to sedated bucks (0/10; P < 0.0001). Similarly, most of does in contact with non-sedated males ovulated (19/20), but none of those in contact with sedated males did so (0/20; P < 0.0001). We conclude that the expression of an intense sexual behavior by male goats is necessary to induce LH preovulatory surge and ovulation in seasonally anovulatory goats.     

The "male effect" in sheep and goats: a review of the respective roles of the two olfactory systems

In sheep and goats, exposure of seasonally anestrous females to sexually active males results in activation of luteinizing hormone (LH) secretion and synchronized ovulation. This phenomenon is named "the male effect" and seems to constitute a major factor in the control of reproductive events. This effect depends mostly on olfactory cues and is largely mimicked by exposure to male fleece only. In sheep, preventing the vomeronasal organ (VNO) from functioning does not affect the female responses to male odor suggesting that, unlike in rodents, the accessory olfactory system does not play the major role in the perception of this pheromonal cue. Female responses also seem to depend on previous experience, an effect that is not common for pheromones and renders this model of special interest. The aim of the present report is to summarize our current knowledge concerning the "male effect" and in particular to clarify the respective roles of the two olfactory systems in the processes involved in this effect.     

Alterations in plasma concentrations of testosterone, LH, and prolactin associated with mating in the male rat.

The hormonal response of the male rat to sexual activity was investigated in two studies. In the first, no evidence of a chronic elevation in plasma levels of testosterone (T), LH, or prolactin (PRL) was observed in sexually experienced rats compared to naive controls. Both groups showed an acute increase in plasma levels of all three hormones following mating, but the increases shown by the experienced group were more pronounced. In the second study, plasma levels of T, LH and PRL rose in sexually experienced male rats following exposure to a mating arena whether it contained an estrous female, an anestrous female, or no other animal. However, the increases were considerably larger in the group exposed to estrous females. It is suggested that plasma hormones rise in anticipation of mating, although not to the same extent as following mating, and that the anticipatory rise may function to initiate or facilitate mating behavior.     

Absence of female-induced luteinizing hormone release in orchidectomized, sexually active mice.

This experiment was designed to determine whether the reflexive LH pulses induced in male mice by exposure to a female are dependent upon the presence of the testes. B6D2F1 males were studied because, unlike most males, these hybrids remain sexually active indefinitely after castration, demonstrating that they still recognize females and are sexually arousable. We reasoned that if LH reflexes occur independently of the testes, then exposing castrated B6D2F1 males to a female should elicit a pulse. We observed female-induced pulses in 67% of intact, naive males and 80% of intact, sexually experienced males, but they were absent in castrated, sexually experienced males. Spontaneous pulses were too frequent in castrated, naive males to distinguish potential LH reflexes. Thus, the induction of LH reflexes depends upon the presence of the testes in these hybrid males. Secondarily we found more frequent spontaneous pulses in sexually naive than in experienced mice regardless of their gonadal status. These unexpected results imply that long-term mating activity slows the pace of spontaneous LH pulses in these hybrids. Considered together, the differential effects of the testes and sexual experience on both types of LH pulses suggest that two distinct pulse-generating mechanisms exist in male mice.     

Male stimulation of luteinizing hormone surge, progesterone secretion and ovulation in spontaneously persistent-estrous, aging rats

In aging, persistently estrous (PE) female rats, there are no estrous cycles or cyclic increases in luteinizing hormone (LH) secretion, but the sexual receptivity to the male is consistently maintained. We recently reported that caging and mating with fertile males elicits an LH surge followed by ovulation in aging PE rats. The present study examined the relationship between the LH surge, the increase in progesterone (P) secretion and ovulation in PE females exposed to males, and assessed whether intromission was essential for the male-induced pre-ovulatory LH surge. PE rats were implanted with intra-atrial cannulae. Six to eight days later, these females were individually caged with a fertile male and repeatedly sampled (once every 30 or 60 min) between 1400 and 1900 h for assays of plasma LH and P. Sexual behavior of the female was recorded and correlated with the changes in plasma LH and P values. Similar experiments were also performed on cannulated PE rats with their vaginal orifice blocked with adhesive tape during the caging and sampling session. In both experiments, over 90% of the PE females displayed a high degree of lordosis response to mounting by the male, and over 60% of those sexually receptive PE females exhibited an LH surge followed by ovulation. The male-induced preovulatory LH surge occurred in PE females without actual intromission. Caging with fertile males also elicited a marked increase in plasma P concentrations in PE rats and in PE females prevented from experiencing intromission.(ABSTRACT TRUNCATED AT 250 WORDS)     

Female effect in sheep. II. Role of volatile substances from the sexually receptive female; implication of the sense of smell

The importance of olfactory cues in inducing luteinizing hormone and testosterone secretion as a response to stimulation by sexually receptive ewes has been tested. In sexually experienced rams, olfactory stimulation with urine, wool and vaginal secretions from sexually receptive females placed in a mask did not induce an endocrine response. The female-induced secretion of LH and testosterone was similar in anosmic, sham-operated and in control rams. These results show that olfactory cues are not necessary in the mediation of interindividual stimulation of endocrine response in the sexually experienced ram.     

Stimulation of reproductive activity in anovulatory Alpine goats exposed to bucks treated only with artificially long days

Two experiments were conducted in a subtropical latitude to determine the response of Alpine male goats to a treatment with artificially long days (experiment 1), and the response of anovulatory lactating Alpine does exposed to males treated only with artificially long days (experiment 2). In experiment 1, one group of males was kept under natural photoperiod (n = 4) while another was exposed to 2.5 months of artificially long days (16 h of light/day) from 1 December (n = 4). Plasma testosterone concentrations were determined weekly. Intensity of odor of males was determined every 2 weeks. Sexual behavior of bucks was observed during 3 days about 90 days after the end of the long day treatment. A treatment-by-time interaction was detected for testosterone secretion (P < 0.001). In control males, low plasma concentrations of testosterone were observed from March to June. In contrast, in long-day treated males, high levels of testosterone were observed from March to June (P < 0.05). A treatment-by-time interaction was detected for the intensity of male odor (P < 0.01). The male odor was stronger in long-day treated bucks than in untreated ones from March to June (P < 0.05). The number of ano-genital sniffing, nudging and flehmen was greater in long-day treated males than in untreated ones when exposed to anestrous does (P < 0.05). In experiment 2, one group of males was left under natural photoperiod variations (n = 5) and the other (n = 5) was submitted to the same photoperiodic treatment described in experiment 1. On 3 May, three untreated and three long-day treated males were put in contact with anestrous Alpine does left under natural photoperiod. Fertility was higher in does exposed to light-treated males (36/45, 80%) than those in contact with untreated ones (3/45, 7%; P < 0.05). Prolificacy was similar (P > 0.05) in does exposed to treated (1.8 ± 0.1) and untreated males (1.7 ± 0.3). These results indicate that the sexual activity of Alpine male goats raised in subtropical latitudes can be induced using only artificially long days and that such males are effective in stimulating reproductive activity in anovulatory females in late spring.     

Kisspeptin Signaling Is Required for the Luteinizing Hormone Response in Anestrous Ewes following the Introduction of Males

The introduction of a novel male stimulates the hypothalamic-pituitary-gonadal axis of female sheep during seasonal anestrus, leading to the resumption of follicle maturation and ovulation. How this pheromone cue activates pulsatile secretion of gonadotropin releasing hormone (GnRH)/luteinizing hormone (LH) is unknown. We hypothesised that pheromones activate kisspeptin neurons, the product of which is critical for the stimulation of GnRH neurons and fertility. During the non-breeding season, female sheep were exposed to novel males and blood samples collected for analysis of plasma LH profiles. Females without exposure to males served as controls. In addition, one hour before male exposure, a kisspeptin antagonist (P-271) or vehicle was infused into the lateral ventricle and continued for the entire period of male exposure. Introduction of a male led to elevated mean LH levels, due to increased LH pulse amplitude and pulse frequency in females, when compared to females not exposed to a male. Infusion of P-271 abolished this effect of male exposure. Brains were collected after the male effect stimulus and we observed an increase in the percentage of kisspeptin neurons co-expressing Fos, by immunohistochemistry. In addition, the per-cell expression of Kiss1 mRNA was increased in the rostral and mid (but not the caudal) arcuate nucleus (ARC) after male exposure in both aCSF and P-271 treated ewes, but the per-cell content of neurokinin B mRNA was decreased. There was also a generalized increase in Fos positive cells in the rostral and mid ARC as well as the ventromedial hypothalamus of females exposed to males. We conclude that introduction of male sheep to seasonally anestrous female sheep activates kisspeptin neurons and other cells in the hypothalamus, leading to increased GnRH/LH secretion.     

Sexual experience interacts with steroid exposure to shape the partner preferences of rats

A three-phase experiment manipulated sexual experience and hormone exposure (perinatally and in adulthood) in female rats housed individually from weaning so as to limit peripubertal social and sexual experience. Noncontact partner preference for a male or estrous female rat was measured both before and after sexual experience, first while rats were under the influence of circulating testosterone propionate (TP) and later after priming them with ovarian hormones (estradiol benzoate and progesterone; EB & P). When implanted with TP capsules and tested while sexually naive, all groups of female rats preferred females to males without differing statistically. However, following three sexual experience sessions with estrous females, differences emerged between the masculinized and control groups in the magnitude of their female-directed preference, with masculinized females demonstrating a significantly greater preference for estrous females. Sexual experience with male rats under EB & P did not result in a significant shift in preference in any group. Histological assessment indicated that the volume of the sexually dimorphic nucleus of the preoptic area (SDN-POA) was increased by exposure to TP postnatally, and SDN-POA volume correlated positively with partner preference scores but only when rats were both sexually experienced and exposed to circulating TP in adulthood. These results suggest that sexual experience interacts with steroid exposure to shape partner preference.     

Hormonal responses to different sexually related conditions in male rats

Plasma levels of corticosterone (C) and testosterone (T) increase after sexual activity in males of several species. However, the physiological significance of these increases has not been elucidated. In the present study, hormonal response to different conditions linked to sexual activity was assessed. In the first experiment, plasma levels of C and T were assessed both in sexually experienced and naive male rats after the following conditions: (A) control group, without sexual stimulation; (B) males exposed to ovariectomized females; (C) males exposed to intact, non-receptive females; (D) males exposed to receptive females with the vagina obstructed, to avoid intromission; (E) males exposed to receptive females: but separated by a grid that prevents physical contact; (F) males exposed to receptive females during 30 min. In a second experiment, experienced male rats were allowed to repeatedly copulate until reaching the criteria for sexual exhaustion, and 24 h later, they were allowed to copulate. Once sexually related conditions ended, males were killed and their blood was obtained. C and T plasma levels were assessed by HPLC with ultraviolet (UV) detection. Results indicate that T did not increase significantly in naive male in any sexual condition, while in the experienced males, significant increases were observed with the mere presence of a receptive female and also after ejaculation. These increases were significantly larger in experienced males. On the other hand, C also increased in all sexual conditions, both in experienced and naive rats; however, the increase observed was larger in experienced males. Regarding sexual satiety, both C and T increased after copulating ad libitum to satiety. T increased almost three-fold compared to control, while C increased two-fold. No significant changes were observed in either one of the steroids 24 h after sexual exhaustion, even though males remained with a receptive female during an hour. These results show that sexual experience has an important influence on the hormonal response to sexual activity. C rises could be directly related to sexual arousal involved in the different sexual conditions, while T rises seem to have a direct relationship with both the motivation and execution aspects of masculine sexual behavior.     

Male Age and Experience Increases Mating Success but Not Female Fitness in the Mexican Fruit Fly

Male age and sexual experience are important condition parameters that can influence mate choice and female fitness, yet they are seldom studied simultaneously. Here, we investigated the effect of male age and previous sexual experience on mating success and female fitness in the Mexican fruit fly, Anastrepha ludens (Diptera: Tephritidae). Males were either young or old, sexually experienced or naïve. Older and sexually experienced males obtained more copulations. However, females did not receive benefits in terms of lifetime fecundity, fertility, or longevity from mating with these males. Results suggest that males become more competitive as they age and gain sexual experience and may be able to maintain a high quality ejaculate compared to young males. Older experienced males may be manipulating females into preferentially mating with them at no benefit to females. Alternatively, females may prefer mating with older more experienced males and possibly receive other indirect benefits, but this remains to be tested.     

The difference of urine between Brandt’s voleLasiopodomys brandtii in the breeding and non-breeding condition

Using animal house experiments we demonstrated that urine odor can elicit physiological changes in the maturation rate of Brandt’s vole (Radde, 1861). We showed that the urine of estrous females, anestrous females and non-breeding males had no effect on the time of sexual maturity of young voles. In contrast, the maturation rate of young female voles was accelerated, and the maturation rate of young male voles was slowed, by exposure to the urine of breeding adult males. Headspace analysis revealed significant differences in the levels of a subset of volatile constituents of urine from breeding and non-breeding males. From a total of 50 components, Propanoic acid, 2,5-Dimethylpyrazine, Dimethyldisulfide, 1-Octene, 1-Hexanol, Hexanoic acid and p-Xylene were observed only in the urine of breeding male Brandt’s voles. All other substances were present in the volatiles from urine of both breeding and non-breeding male Brandt’s vole.