Copulatory Behavior as Courtship Communication

In the copulatory behavior of many species of rodents, mounting patterns with ejaculation are preceded by mounts with nonejaculatory intromissions. Although not directly related to insemination, these intromissions have generally been treated as “copulatory” behavior. If we treat these nonejaculatory intromissions as courtship communication in the tactile modality, new relationships become apparent. Within the context of classical ethology, intromissions can be considered in relation to both proximate dynamics and ultimate effects. Within the sociobiological/behavioral-ecological context, intromissions can be viewed in relation to manipulation, female assessment and choice, and honesty of communication. The intromissions synchronize, persuade, and orient at the same time as they manipulate the partner.     

Testosterone induction of male-typical sexual behavior is associated with increased preoptic NADPH diaphorase and citrulline production in female whiptail lizards

In rodents, male-typical copulatory behavior is generally dependent on gonadal sex steroids such as testosterone, and it is thought that the mechanism by which the hormone gates the behavior involves the gaseous neurotransmitter nitric oxide. According to one model, testosterone induces an up-regulation of nitric oxide synthase (NOS) in the preoptic area, increasing nitric oxide synthesis following exposure to a sexual stimulus. Nitric oxide in turn, possibly through its effect on catecholamine turnover, influences the way the stimulus is processed and enables the appropriate copulatory behavioral response. In whiptail lizards (genus Cnemidophorus), administration of male-typical levels of testosterone to females induces the display of male-like copulatory responses to receptive females, and we hypothesized that this radical change in behavioral phenotype would be accompanied by a large change in the expression of NOS in the preoptic area. As well as comparing NOS expression using NADPH diaphorase histochemistry between testosterone-treated females and controls, we examined citrulline immunoreactivity (a marker of recent nitric oxide production) in the two groups, following a sexual stimulus and following a nonsexual stimulus. Substantially more NADPH diaphorase-stained cells were observed in the testosterone-treated animals. Citrulline immunoreactivity was greater in testosterone-implanted animals than in blank-implanted animals, but only following exposure to a sexual stimulus. This is the first demonstration that not only is NOS up-regulated by testosterone, but NOS thus up-regulated is activated during male-typical copulatory behavior.     

Testosterone induction of male‐typical sexual behavior is associated with increased preoptic NADPH diaphorase and citrulline production in female whiptail lizards

In rodents, male‐typical copulatory behavior is generally dependent on gonadal sex steroids such as testosterone, and it is thought that the mechanism by which the hormone gates the behavior involves the gaseous neurotransmitter nitric oxide. According to one model, testosterone induces an up‐regulation of nitric oxide synthase (NOS) in the preoptic area, increasing nitric oxide synthesis following exposure to a sexual stimulus. Nitric oxide in turn, possibly through its effect on catecholamine turnover, influences the way the stimulus is processed and enables the appropriate copulatory behavioral response. In whiptail lizards (genus Cnemidophorus), administration of male‐typical levels of testosterone to females induces the display of male‐like copulatory responses to receptive females, and we hypothesized that this radical change in behavioral phenotype would be accompanied by a large change in the expression of NOS in the preoptic area. As well as comparing NOS expression using NADPH diaphorase histochemistry between testosterone‐treated females and controls, we examined citrulline immunoreactivity (a marker of recent nitric oxide production) in the two groups, following a sexual stimulus and following a nonsexual stimulus. Substantially more NADPH diaphorase‐stained cells were observed in the testosterone‐treated animals. Citrulline immunoreactivity was greater in testosterone‐implanted animals than in blank‐implanted animals, but only following exposure to a sexual stimulus. This is the first demonstration that not only is NOS up‐regulated by testosterone, but NOS thus up‐regulated is activated during male‐typical copulatory behavior. © 2006 Wiley Periodicals, Inc. J Neurobiol, 2006     

Differences in responsiveness to testosterone of penile reflexes and copulatory behavior of male rats

The display of penile reflexes and copulatory behavior appears to reflect the activity of two different underlying neuronal system, both of which are modulated by systemic testosterone (T) concentration. To indirectly compare the two systems, the responsiveness to T of penile reflexes and copulatory behavior was examined. In the first experiment castrated spinal male rats were given penile reflex tests while receiving replacement T through Silastic capsule implants filled with T (50 mm T). After capsule removal the number of penile erections and flips declined within 24 hr and gradually decreased for 12 days. Subjects were then reimplanted with new 50-mm T capsules. The number of penile flips and erections increased within 6 and 12 hr. respectively. This is a much more rapid response rate to T than has been established for copulatory behavior. In the second experiment castrated spinal male rats were tested for penile reflexes with a 50-mm T capsule, which was then replaced with a 10-, 5-, or 2-mm T or an empty capsule. The number of penile reflexes declined in a dose-response fashion. In the third experiment, castrated sexually experienced male rats were tested for copulatory behavior with two 25-mm T capsules which were then replaced with a 10 or 2-mm T or an empty capsule. Only males with empty capsules had decrements in copulatory behavior, revealing that a low level of T can maintain virtually normal sexual behavior despite a marked decline in penile reflex activity. The neuronal system underlying penile reflexes (spinal neurons) is apparently much more responsive to changes in T concentrations than the neuronal system underlying motivational and appetitive aspects of copulatory behavior (brain neurons).     

Sex differences and effects of neonatal aromatase inhibition on masculine and feminine copulatory potentials in prairie voles

Copulatory behaviors in most rodents are highly sexually dimorphic, even when circulating hormones are equated between the sexes. Prairie voles (Microtus ochrogaster) are monomorphic in their display of some social behaviors, including partner preferences and parenting, but differences between the sexes in their masculine and feminine copulatory behavior potentials have not been studied in detail. Furthermore, the role of neonatal aromatization of testosterone to estradiol on the development of prairie vole sexual behavior potentials or their brain is unknown. To address these issues, prairie vole pups were injected daily for the first week after birth with 0.5 mg of the aromatase inhibitor 1,4,6-androstatriene-3,17-dione (ATD) or oil. Masculine and feminine copulatory behaviors in response to testosterone or estradiol were later examined in both sexes. Males and females showed high mounting and thrusting in response to testosterone, but only males reliably showed ejaculatory behavior. Conversely, males never showed feminine copulatory behaviors in response to estradiol. Sex differences in these behaviors were not affected by neonatal ATD, but ATD-treated females received fewer mounts and thrusts than controls, possibly indicating reduced attractiveness to males. In other groups of subjects, neonatal ATD demasculinized males' tyrosine hydroxylase expression in the anteroventral periventricular preoptic area, and estrogen receptor alpha expression in the medial preoptic area. Thus, although sexual behavior in both sexes of prairie voles is highly masculinized, aromatase during neonatal life is necessary only for females' femininity. Furthermore, copulatory behavior potentials and at least some aspects of brain development in male prairie voles are dissociable by their requirement for neonatal aromatase.     

A comparative description of the mating behaviour of female rodents

This paper provides a comparative analysis of movements and postures of female rodents during copulation. Cinematographic techniques were used to assess details of movements and postures of the copulatory behaviour of laboratory (Norway) rats, Mongolian gerbils, and 14 other species of muroid rodents. Considerable variability, with potential evolutionary, ecological, and physiological implications, was found.     

Organizational effects of estrogens on brain vasotocin and sexual behavior in quail

Reproductive behavior is sexually differentiated in quail: The male-typical copulatory behavior is never observed in females even after treatment with high doses of testosterone (T). This sex difference in behavioral responsiveness to T is organized during the embryonic period by the exposure of female embryo to estrogens. We showed recently that the sexually dimorphic medial preoptic nucleus (POM), a structure that plays a key role in the activation of male copulatory behavior, is innervated by a dense steroid-sensitive network of vasotocin-immunoreactive (VT-ir) fibers in male quail. This innervation is almost completely absent in the female POM and is not induced by a chronic treatment with T, suggesting that this neurochemical difference could be organizational in nature. This idea was tested by injecting fertilized quail eggs of both sexes on day 9 of incubation with either estradiol benzoate (EB) (25 μg, a treatment that suppresses the capacity to show copulatory behavior in adulthood) or the aromatase inhibitor R76713 (10 μg, a treatment that makes adult females behaviorally responsive to T), or with the solvents as a control (C). At 3 weeks posthatch, all subjects were gonadectomized and later implanted with Silastic capsules filled with T. Two weeks later, all birds were perfused and brain sections were processed for VT immunocytochemistry. Despite the similarity of the adult endocrine conditions of the subjects (all were gonadectomized and treated with T Silastic implants providing the same plasma level of steroid to all subjects), major qualitative differences were observed in the density of VT-ir structures in the POM of the different groups. Dense immunoreactive structures (fibers and a few cells) were observed in the POM of C males but not females; EB males had completely lost this immunoreactivity (and lost the capacity to display copulatory behavior); and, conversely, R76713 females displayed a male-typical VT-ir system in the nucleus (and also high levels of copulatory behavior). Similar changes in immunoreactivity were seen in the nucleus of the stria terminalis and in the lateral septum (VT-ir fibers only in this case) but not in the magnocellular vasotocinergic system. These neurochemical changes closely parallel the effects of the embryonic treatments on male copulatory behavior. The vasotocinergic system of the POM can therefore be considered an accurate marker of the sexual differentiation of brain circuits mediating this behavior. © 1998 John Wiley & Sons, Inc. J Neurobiol 37: 684–699, 1998     

The Influence of Male Body Size and Social Environment on the Mating Behavior of Phallichthys quadripunctatus (Pisces: Poeciliidae)

In some poeciliid fishes, variation in male size is accompanied by differences in mating behavior. Large males are preferred by females and perform courtship displays followed by copulatory thrusts, whereas small males perform copulatory thrusts with few or no displays. This phenomenon has been described in only a few genera and little is known about mating behavior in other poeciliids. Although Phallichthys quadripunctatus males display size dimorphism that has a genetic component, mating behavior of this species has not been documented. We conducted experiments using socially experienced and socially naive males to characterize the mating behavior of this species and to evaluate potential size-dependent differences in behavior. Males were tested with postpartum (presumably receptive) and midcycle (presumably unreceptive) females in different social environments. Whereas neither size class of P. quadripunctatus males performed courtship displays or altered behavior based on female receptivity, large males performed several reproductive behaviors more frequently than small males. This trend was repeatable and occurred in all social environments examined. Some males also attempted to mate with other males, with small males showing a greater tendency to perform this behavior than large males. The manner in which differences in reproductive activity translate into differences in reproductive success must be examined before inferring sexual selection favoring large males in P. quadripunctatus.     

外源性皮质酮对雄性根田鼠交配行为的作用

本文研究了皮质酮对雄性根田鼠交配行为的作用。实验个体分别注射0.10 ug/ g体重、0. 60 ug / g体重和1.00 ug/ g体重剂量的皮质酮,1 h后测定每只雄性根田鼠的交配行为。结果显示,3 个处理组动物具有射精能力的个体比率及其爬跨、抽动和射精潜伏期与对照组动物相比均无显著差异,爬跨和抽动频次也无显著变化。不同处理组个体的血浆睾酮含量无显著差异。因此,皮质酮没有影响雄性根田鼠的交配行为和性激素的分泌。该结果提示,哺乳动物在急性应激条件下所分泌的皮质酮可能不参与对性行为的调控。     

Female Meadow Voles, Microtus pennsylvanicus, Experience a Reduction in Copulatory Behavior During Postpartum Estrus

In many small mammalian species, females undergo postpartum estrus (PPE), mate, and become pregnant soon after delivering a litter. Mating during PPE implies a trade‐off between attending to the new litter and leaving that litter to mate. We tested the hypothesis that copulatory behavior is faster when a female mates during PPE than when it mates outside of PPE, during male‐induced estrus (MIE), a time when it is not lactating. We compared several variables of copulatory behavior in female meadow vole mating during both PPE and MIE. Females in PPE received significantly fewer intromissions, with shorter intervals between intromissions. Each ejaculatory series was also shorter for females in PPE. As a consequence, the total amount of time devoted to copulation was much shorter for females in PPE than for those in MIE. These data support the hypothesis that female meadow voles are able to reduce their copulatory behavior during PPE.     

Decline of sexual behavior in castrated male rats: Effects of female precopulatory behavior

From the population of 89 adult sexually inexperienced Wistar male rats 20 animals that initiated copulatory behavior with females exhibiting low intensity of precopulatory behavior (presenting females) were preselected. Prior to castration all 20 males had the same sexual experience: three ejaculatory series in four weekly sessions with females exhibiting high intensity of precopulatory behavior (darting females). Following castration, the decline of copulatory behavior was much slower for the nine males tested with darting females as compared to the 11 males tested with presenting females. Male precopulatory behaviors (anogenital sniffing, touching flanks, etc) outlasted the loss of copulatory behavior and seem to be less dependent on both external and internal determinants. It is concluded that intensive external sexual stimuli can function to compensate, and therefore mask, the subnormal operation of androgen-dependent mechanisms in initiating the copulatory behavior.     

Effects of i.c.v. administration of leptin on copulatory and ingestive behavior in STZ-induced diabetic male rats.

It is well known that circulating leptin concentrations correlate with adiposity in both humans and rodents and decrease after fasting, energy restriction, or weight loss. The goal of the present study was to confirm whether the decreases of copulatory behavior and the increases of ingestive behavior in STZ-induced diabetic male rats could be reversed by i.c.v. administration of leptin. Adult male Wistar-Imamichi rats aged 9 weeks were used for the studies. Males received a single injection of STZ (60 mg/kg, i.p.) and vehicle. During the experiment, individual body weight, and food and water intake were measured. The copulatory and ingestive behaviors in STZ-induced diabetic males were observed at 2 and 4 weeks after STZ. At 6 weeks after STZ, leptin (10 microg/10 microl) or aCSF (artificial cerebrospinal fluid) was injected through a lateral ventricle cannula and the above two behaviors were observed again. The i.c.v. leptin injection to STZ-induced diabetic males resulted in a significant increase of ejaculation frequencies (3.6 +/- 0.26 vs. 2.9 +/- 0.30 times) and a significant decrease in amount of food ingested (36.2 +/- 1.93 vs. 23.2 +/- 3.76 g), compared with the aCSF-injected control (p<0.01). These findings suggest that the copulatory and ingestive behaviors in i.c.v. leptin-injected STZ diabetic males were restored to levels equivalent to those in control males.     

Sexual differentiation of brain and behavior in quail and zebra finches: Studies with a new aromatase inhibitor, R76713

In many species of vertebrates, major sex differences affect reproductive behavior and endocrinology. Most of these differences do not result from a direct genomic action but develop following early exposure to a sexually differentiated endocrine milieu. In rodents, the female reproductive phenotype mostly develops in the absence of early steroid influence and male differentiation is imposed by the early action of testosterone, acting at least in part through its central conversion into estrogens or aromatization. This pattern of differentiation does not seem to be applicable to avian species. In Japanese quail (Coturnix japonica), injection of estrogens into male embryos causes a permanent loss of the capacity to display male-type copulatory behavior when exposed to testosterone in adulthood. Based on this experimental result, it was proposed that the male reproductive phenotype is “neutral” in birds (i.e. develops in the absence of endocrine influence) and that endogenous estradiol secreted by the ovary of the female embryo is responsible for the physiological demasculinization of females. This model could be recently confirmed. Females indeed display a higher level of circulating estrogens that males during the second part of their embryonic life. In addition, treatment of female embryos with the potent aromatase inhibitor, R76713 or racemic vorozole™ which suppresses the endogenous secretion of estrogens maintains in females the capacity to display the full range of male copulatory behaviors. The brain mechanisms that control this sexually differentiated behavior have not been identified so far but recent data suggest that they should primarily concern a sub-population of aromatase-immunoreactive neurons located in the lateral parts of the sexually dimorphic preoptic nucleus. The zebra finch (Taeniopygia guttata) exhibits a more complex, still partly unexplained, differentiation pattern. In this species, early treatment with exogenous estrogens produces a masculinization of singing behavior in females and a demasculinization of copulatory behavior in males. Since normal untreated males sing and copulate, while females never show these behaviors even when treated with testosterone, it is difficult to understand under which endocrine conditions these behaviors differentiate. In an attempt to resolve this paradox, we recently treated young zebra finches with R76713 in order to inhibit their endogenous estrogens secretion during ontogeny and we subsequently tested their behavior in adulthood. As expected, the aromatase inhibitor decreased the singing frequency in treated males but it did not affect the male-type copulatory behavior in females nor in males. In addition, the sexuality differentiated brain song control nuclei which are also masculinized in females by early treatment with estrogens, were not affected in either sex by the aromatase inhibitor. In conclusion, available data clearly show that sexual differentiation of reproductive behaviors in birds follows a pattern that is almost opposite to that of mammals. This difference may be related to the different mechanisms of sex determination in the two taxa. In quail, the ontogeny of behavioral differentiation is now well understood but we only have a very crude notion of the brain structures that are concerned. By contrast, in zebra finches, the brain mechanisms controlling the sexually differentiated singing behavior in adulthood have been well identified but we do not understand how these structures become sexually dimorphic during ontogeny.     

Copulation behavior inside and outside the beetleMacrohaltica jamaicensis (Coleoptera,Chrysomelidae)

Copulation behavior inMacrohaltica jamaicensis involved a complex, relatively stereotyped series of events both inside and outside the female. Only incomplete sperm transfer occurred in many copulations. Some aspects of male copulatory courtship behavior performed during and following copulation were consistently coordinated with particular stages in sperm transfer. This coordination suggests that copulatory courtship may function to induce critical internal female responses which sometimes do not occur, such as relaxing the bursal muscles to permit deeper penetration, and allowing sperm transfer to occur before the spermatophore is discarded.     

Sperm competition and diversity in rodent copulatory behaviour

The copulatory behaviour of male mammals is characterized by striking diversity in patterns of copulatory stimulation and ejaculation frequency. We conducted comparative analyses of rodents to investigate the potential influence of sperm competition in the evolution of copulatory behaviour. We found that increasing sperm competition is associated with faster rates of stimulation and earlier ejaculation among species in which males perform multiple intromissions prior to ejaculation, but with no overall change in the number of intromissions per ejaculation. Increasing sperm competition is also associated with a higher frequency of repeated ejaculations with the same female, and with shorter refractory periods between repeated copulations. Increasing sperm competition level thus appears to select for male ability to ejaculate more rapidly and more frequently with each female mated. As prolonged copulations are known to reduce female remating rates, these findings indicate that males may experience opposing selection pressures on copulatory behaviour with respect to offensive and defensive aspects of sperm competition. We conclude that sperm competition is likely to be an important selection pressure explaining diversity in mammalian copulatory behaviour.     

Sexual differentiation in quail: Conversion of androgen to estrogen mediates testosterone-induced demasculinization of copulation but not other male characteristics

Previous research has shown that administration of either testosterone or estradiol to male quail embryos will demasculinize behavior and morphology. Six experiments in which embryos were treated were conducted to test the hypothesis that this testosterone-induced demasculinization is due to conversion of testosterone to estrogen (aromatization). In Experiment 1, dihydrotestosterone propionate, a nonaromatizable androgen, failed to demasculinize copulatory behavior, but did demasculinize crowing, strutting, and proctodeal glands. In Experiment 2, injection of the aromatizable androgens testosterone propionate (TP), testosterone, or androstenedione demasculinized copulatory behavior, the nonaromatizable androgen androsterone failed to have such an effect, and all androgens demasculinized proctodeal glands. In Experiment 3, Silastic implants of testosterone demasculinized all male characteristics, whereas implants of androsterone demasculinized only proctodeal glands. In Experiment 4, the antiestrogen tamoxifen prevented TP from demasculinizing copulatory behavior, but had no such effect with respect to crowing and strutting. In Experiments 5 and 6, the aromatization inhibitor 1,4,6-androstatrien-3,17-dione (ATD) prevented TP but not estradiol benzoate from demasculinizing copulatory behavior. Thus (1) in quail, testosterone-induced demasculinization of copulatory behavior is due to androgen aromatization, whereas testosterone-induced demasculinization of crowing, strutting, and proctodeal glands is not; (2) the distinct components of normal male reproductive behavior exhibit different patterns of steroid specificity during the organizational period, as was previously shown for the activational period; (3) the steroid specificity of crowing, strutting, and proctodeal glands changes between the organizational and activational periods. During organization, there is little specificity, whereas during activation, these characteristics respond only to androgens, never to estrogens. This difference suggests that developmental changes have occurred in the underlying biochemical substrates.     

Brief refeeding restores reproductive readiness in food-restricted female musk shrews (Suncus murinus)

Food deprivation blocks sexual behavior and disrupts estrous cycles in mammals. We asked whether reduced copulatory behavior, produced by limited food intake, could be reversed by brief refeeding intervals in the female musk shrew. In Experiment 1, animals were food restricted to 60% of ad lib (FR), and an additional group of FR females were refed for 90 min prior to testing (RF). Refed and ad lib (AL) fed females were significantly more likely to mate than FR females. To test the hypothesis that food-induced restoration of copulatory behavior is not the result of changes in peripheral steroids, we repeated Experiment 1 using ovariectomized and testosterone-implanted females. The results from Experiment 2 were similar to those found in the first study. Next, a more severe refeeding schedule was employed; females were restricted to 50% of ad lib intake. Females in the RF and FR groups were significantly less likely than the AL animals to mate. In the last experiment, females were food restricted to 50% and longer refeeding intervals were employed. Four and one-half hours of food intake did not reinstate sexual behavior, but females refed for 12 h were as likely to mate as ad lib fed controls. We also did not detect any differences in plasma concentrations of testosterone and cortisol in AL, FR, and RF ovary-intact animals. These results define a nutritional threshold for copulatory behavior in the musk shrew. Since this species is highly sensitive to small alterations in food intake, it is a useful model for studies of interactions between metabolic fuels and behavior.     

Gonadal influences on sexual behavior in the male musk shrew (Suncus murinus)

In this series of experiments the hormonal bases for male copulatory behavior in the musk shrew (Suncus murinus) were examined. Male musk shrews failed to show copulatory behavior after castration. Testosterone replacement fully reinstated sexual behavior. Males castrated at birth, and tested as adults after receiving testosterone implants, did not show male-typical sexual behavior. It appears that the gonads are essential for the regulation of male sexual behavior in this primitive mammal.     

Copulatory behavior of sexually inexperienced male voles paired with proestrous females

The copulatory behavior in sexually inexperienced male voles aged 9-13 weeks old was observed under dim red illumination. Proestrous females were used as stimulus voles in copulatory behavior tests. A 30-min test session was recorded. All males showed at least one ejaculation within 30 min. In addition, the pattern of copulatory behavior in male voles was compared to that reported previously in male hamsters. The frequencies of mount, intromission and ejaculation in male voles were similar to that in male hamsters but there were significant differences in the latencies of mounting, intromission, ejaculation and post-ejaculation between the two species.     

Copulatory behavior of sexually inexperienced male hamsters paired with natural proestrous females

The copulatory behavior of sexually inexperienced male hamsters paired with natural proestrous females was observed between 1900 and 2000 hours under dim red light illumination. Each male was allowed 30 min to mate with a receptive female. Five out of 7 males showed at least one ejaculation within 30 min. Although the copulatory behavior of the above males was observed again, the activities of copulatory behavior did not facilitate.