Conditioned odor aversion induces social anxiety towards females in wild‐type and TrpC2 knockout male mice

Female‐emitted pheromonal inputs possess an intrinsic rewarding value for conspecific males, promoting approach and investigation of the potential mating partner. In mice these inputs are detected mainly by the vomeronasal organ (VNO) and the main olfactory epithelium (MOE). We investigated the role of VNO‐mediated inputs in experience‐dependent plasticity of reproductive responses. We applied a sex‐specific conditioned odor aversion (COA) paradigm on adult, wild‐type (WT) male mice and on male mice impaired in VNO‐mediated signal transduction (TrpC2^?/?). We found that WT males, which underwent COA to female‐soiled bedding, lost their innate preference to female odors and presented lower motivation to approach a sexually receptive female. COA also abolished the testosterone surge normally seen following exposure to female odors. Moreover, the conditioned males displayed impairments in copulatory behaviors, which lasted for several weeks. Surprisingly, these males also exhibited phobic behaviors towards receptive females, including freezing and fleeing responses. In contrast, WT males which underwent COA specifically to male pheromones showed no change in olfactory preference and only a marginally significant elevation in intermale aggression. Finally, we show that TrpC2^?/? males were able to acquire aversion to female‐soiled bedding and presented similar behavioral alterations following COA in their responses to female cues. Our results demonstrate that the intrinsic rewarding value of female pheromones can be overridden through associative olfactory learning, which occurs independently of VNO inputs, probably through MOE signaling.     

Social motivation is reduced in vasopressin 1b receptor null mice despite normal performance in an olfactory discrimination task

In this study, we characterized more thoroughly the social behavior of vasopressin 1b receptor null (V1bR-/-) mice. We confirmed that V1bR-/- males exhibit less social aggression than their wild-type (V1bR+/+) littermates. We tested social preference by giving male subjects a choice between pairs of soiled or clean bedding. In general, V1bR+/+ mice spent significantly more time engaged in chemoinvestigation of these social stimuli than V1bR-/- mice. Male V1bR+/+ mice preferred female-soiled bedding over male-soiled bedding, male-soiled bedding over clean bedding, and female-soiled bedding over clean bedding. In contrast, V1bR-/- males failed to exhibit a preference for any bedding. This difference in behavior is not explained by an anosmic condition as there were no differences between V1bR-/- and V1bR+/+ mice in their abilities to detect a cookie buried in clean bedding, or in their ability to perform in an operant conditioning task using a fully automated liquid dilution olfactometer. In the latter task, male V1bR-/- mice were fully capable of discriminating between male and female mouse urine. The latencies to learn this task did not differ between the two genotypes. Thus, a V1bR-/- male's ability to differentiate between male and female chemosensory cues appears no different than that of a V1bR+/+ male's. We propose that the V1bR plays an important role in social motivation, perhaps by coupling the processing, integration, and/or interpretation of chemosensory cues with the appropriate behavioral response.     

Context alters the ability of clitoral stimulation to induce a sexually-conditioned partner preference in the rat

We have shown previously that clitoral stimulation (CLS) of female rats induces significant conditioned place preference (CPP), indicating that it is rewarding. The present study asked whether CLS could induce a conditioned partner preference. In the first experiment, sexually naïve females received 10 alternating trials of CLS and No-CLS in the presence of a male rat behind a wire-mesh screen. For one group, CLS was made in the presence of the male scented with almond extract. On alternating trials, those females received sham CLS in the presence of an unscented male behind the screen. The order was reversed for the other group. After 5 trials in each condition, females were placed into an open field with two sexually vigorous males, one scented and the other unscented. Contrary to expectation, females displayed a preference for the male associated with sham CLS. The second experiment examined whether a partner preference could be conditioned by associating CLS with the almond odor alone. A new group of sexually naive females received the same CLS-odor, No-CLS-No Odor pairings as above, but with the odor presented on cotton gauze in the chamber. During the final open field test, those females selectively solicited the scented male. We conclude that CLS that induces CPP also induces conditioned partner preference. However, we propose that CLS in the presence of an inaccessible male created a sexual inhibitory state for female rats.     

Subspecies recognition in the house mouse: a study of two populations from the border of a hybrid zone

Mate choice is the outcome of sexual preference for partnerscarrying specific signals. Thus, mating among conspecifics(homogamy) depends on the occurrence of species recognitionsystems. We asked what happens if populations diverge, andwe investigated female sexual preference between two subspeciesof the house mouse in populations from the borders of a hybridzone (Jutland, Denmark). We used choice tests to analyze theoccurrence of recognition signals and to locate these signalsin soiled bedding and urine. Our results show that populationsof the two subspecies can be discriminated on the basis ofurinary signals, suggesting that the latter have diverged. Additionally, these signals seem to have similar features inpopulations of different geographical origins, suggesting thatsubspecific differentiation occurs. This is the first demonstrationthat subspecific recognition through urinary signals occursin the house mouse. However, while Mus musculus domesticusdoes not display a preference, we show that Mus musculus musculusfemales tend to mate with males of the same subspecies. We discuss the different factors that could explain these discrepanciesbetween females of the two taxa: differences in signal perception,evolution at a different pace, or evolution under differentselective pressures in their area of contact. Further, we proposethat the divergence in male signal was at least partly initiatedin allopatry and discuss different evolutionary scenario thatmay explain the patterns observed in Denmark and their relevance to isolation between the two taxa.     

Population and Species Divergence of Chemical Cues that Influence Male Recognition of Females in Desmognathine Salamanders

Growing evidence indicates that males may be more discriminating of mating partners than often has been assumed. In the North American Ocoee dusky salamander, Desmognathus ocoee (Plethodontidae: Desmognathinae), sexual incompatibility among conspecific populations is high in encounters staged in the laboratory, at least in part because males fail to recognize ‘other’ females as appropriate targets for courtship. I used Y-mazes to test the hypothesis that males of D. ocoee discriminate between substrate-borne chemical cues produced by ‘own’ (homotypic) and ‘other’ (heterotypic) females. Males of four populations discriminated in favor of substrates soiled by homotypic females over clean (control) substrates (expt 1), suggesting that females produce chemical cues of sociosexual significance to males. Furthermore, males from these populations discriminated in favor of substrates soiled by homotypic females vs. substrates soiled by heterotypic females (expt 2), both conspecific and heterospecific (D. carolinensis and D. orestes). Thus, differences among populations and species in female chemical cues appear to affect the chemotactic responses of males. I suggest that, together with differences in behavioral signals and responses exhibited during courtship, differences in female chemical cues likely contribute to sexual incompatibility among populations and taxa of desmognathine salamanders.     

Role of photoperiod and temperature in production of the oestrus-inducing pheromone in male wild mice

Ninety adult males divided in six equal groups were exposed to different photoperiods for 21 days. Exposures included natural light (ca 11 hr), long photoperiod (16L:8D) and short photoperiod (8L:16D). The first three groups received these exposures at room temperature (13-20 degrees C) while the remaining three at raised temperature (36-38 degrees C). Soiled bedding of the above males was introduced in the cages of unisexually housed noncyclic females and their potentiality to induce oestrus was assessed. It was noticed that the bedding of all the males proved to be a stimulus inducing oestrus in the majority of the females during the 7 day exposure. There was no significant difference in the number of females returning to oestrus following exposure to soiled bedding of different males. These results elucidate that environmental factors, especially light and temperature do not influence the production/release of the oestrus-inducing pheromone in wild mice.     

Peripubertal exposure to male odors influences female puberty and adult expression of male-directed odor preference in mice

Testosterone-dependent olfactory signals emitted by male are well known to accelerate female puberty in mice (Vandenbergh effect). However, it remains unclear whether these chemosignals also influence adult expression of male-directed odor preference. Therefore, we exposed female mice to intact or castrated male bedding (vs clean bedding as control) during the peripubertal period (postnatal day (PD) 21–38) and measured male-directed odor preference in adulthood. At PD45 or PD60, females exposed to intact male odors, and thus showing puberty acceleration, preferred to investigate odors from intact males over females or castrated males. Females exposed to castrated male odors did not show puberty acceleration but preferred male (intact or castrated) over female odors. Finally, control females did not show any odor preference when tested at PD45, although a preference for male odors emerged later (PD60). In a second experiment, females that were exposed to intact male odors after pubertal transition (PD36–53) also preferred intact male over castrated male odors. In conclusion, our results indicate that peripubertal exposure to male odors induced early expression of male-directed odor preference regardless of puberty-accelerating effect and that induction of male-directed odor preference is not specific to the peripubertal period.     

Behavioural development of conspecific odour preferences in Brandt’s voleMicrotus brandti

The behavioural development of conspecific odour preferences in Brandt’s voleMicrotus brandti (Radde, 1861) was evaluated in laboratory tests. During the weaning period (15th-30th day after the birth), the preference of pups to individual odours of their parents, novel adult males and females were examined. Pups investigated odours of unfamiliar adults significantly more than those of their parents and they displayed significantly more jumping and upward visual investigating behaviours on the bedding soiled by unfamiliar adults than they did on the bedding of their parents. However, the frequency of crossing the parent’s bedding was significantly higher than the frequency of crossing the bedding of alien adults. It is suggested that odour discrimination ability was associated with the age of pups. The Brandt’s vole pups might keep themselves away from unfamiliar environments by keeping away from the novel conspecific odours.     

Only Self-Paced Mating Is Rewarding in Rats of Both Sexes

When rats are mated in a traditional mating chamber (with one male and one female) in which the male dictates the pace of the copulatory sequence, males develop a reward state as evaluated by conditioned place preference (CPP). In this mating situation no reward state is induced in females. However, when female rats are able to control (pace) the rate of sexual stimulation, thereby reducing the aversive consequences associated with mating, a clear CPP is observed. In the present study the CPP paradigm was used to determine whether if the reinforced state induced by coital interactions in male rats can be maintained when females pace the sexual interaction. Adult male and female rats were mated in one of two different conditions: (1) where subjects were able to pace their coital interactions or (2) where subjects were not able to pace their sexual contacts. The results showed that when males had control over the sexual interaction they developed a clear place preference while males that mated with females that paced their coital contacts did not develop CPP. Similarly, only females that were able to pace their sexual contacts developed place preference. These results suggest that coital interactions in males, as well as in females, can induce a reward state only when they are able to control the sexual interaction. Under seminatural conditions sexual behavior in rats is highly promiscuous, they mate in groups and repeatedly change partners in the middle of copulation. This behavioral sequence allows both, male and female to control the rate of sexual interaction, assuring the induction of a reward state outlasting the actual performance of coital responses.     

Intermittent stimulation and delay of puberty by urinary chemosignals and social contact in wild stock female house mice (Mus Domesticus)

A sequence of six experiments using wild stock house mouse (Mus domesticus) tested the effects of intermittent stimulation with either the urinary chemosignal released by grouped female mice or social contact from grouped females on the age of first vaginal oestrus in young females. Weanling female mice were exposed to bedding soiled by grouped females or cages containing grouped females for 15 min periods, then removed for a prescribed period, and placed again in a cage with soiled bedding or grouped females. The nature of the exposure to the puberty delaying effect, the number of total exposures each day, the total length of exposure to the stimulus, and the total time period over which the exposures occurred were varied. None of the treatment regimes employed here with soiled bedding from grouped females resulted in delays in the onset of first oestrus in test females. Young females exposed to grouped females for 6 or 8 exposures in a 4 h period, 6 or 8 exposures in an 8 h period, or 8 exposures in a 12 h period were significantly delayed in attaining puberty relative to control females that were exposed to cages containing clean bedding. These results are in contrast to earlier findings involving chemosignals that accelerate first oestrus wherein young females exhibited the capacity to accumulate the exposures to the urinary chemosignals from males, females in oestrus and pregnant or lactating females. Direct exposure to the grouped females on an intermittent basis can provide stimulation that is cummulative and results in delays in the onset of first oestrus.     

Species-specific releaser effect of urine from ovulated female masu salmon and rainbow trout

MT (17α-methyltestosterone)-treated immature masu salmon Oncorhychus masou and rainbow trout Oncorhynchus mykiss , exposed to the urine of ovulated females in their non-reproductive season, made significantly more entries into the channel, of a Y-maze preference trough, conditioned by the urine of conspecific females than by heterospecifics. Immature, non-treated controls in both species showed no preference for the urine of ovulated females. These results suggest that, for both species, the releaser pheromones that attract males in the urine are species-specific. Therefore, utilization of species-specific chemical signals may be one of the mechanisms of the reproductive isolation in Oncorhynchus species.     

Oestrous cycle disruption in group-housed mice: evaluation of the involvement of tactile and pheromonal stimuli.

The role of tactile stimuli and pheromonal stimuli in the induction of oestrous cycle irregularities in mice was evaluated. In contrast to an adult regularly cycling female (the test female) housed in contact with 6 adult females, a test female housed in contact with 6 impuberal females failed to show disruption of oestrous cycle. Likewise, a test female housed in contact with bedding soiled by 6 group-housed adult females failed to exhibit disruption of oestrous cycle. By contrast, a test female housed in contact with 6 impuberal females on bedding soiled by 6 adult group-housed females exhibited a significant increase in the incidence of prolonged cycles. However, a test female housed with 6 impuberal females without direct contact with the bedding soiled by adult group-housed females exhibited a significant decrease in the incidence of prolonged cycles. The results suggest that tactile stimuli and pheromonal stimuli (probably present in the excrete of adult females) act synergistically in inducing cycle disruption. The findings further indicate that the female-originating pheromone involved in oestrous cycle disruption is non-volatile (not air-borne) and that it acts through contact.     

The influence of pheromones produced by freely growing laboratory populations on the reproductive maturation of prairie deermice following prenatal and postnatal exposure

Summary Significantly more females reared on bedding soiled by different populations reproduced by 100 days of age than those reared on bedding soiled by the same populations (P<.1) or on clean bedding (P<.025) whereas the latter two groups did not differ significantly. Significantly greater percentages of females reproduced in both treatments maintained on soiled bedding than was true in the populations from which the bedding came. There were no significant differences between treatments in the weights of the reproductive organs (testes, seminal vesicles, ovaries, uteri) or in the numbers of corpora lutea or mature follicles in the ovaries of nulliparous females. The data suggest that if there were reproductively inhibiting pheromones influencing the population animals, they were altered or dissipated immediately after removal of the bedding from the population enclosures or that they were nonfunctional in the absence of the population animals. These data further suggest that the reproductive inhibition of population young is due to the presence of the populaton animals and that sociobiological factors within the populations act to prevent the reproductively stimulating influence of materials which may be present.     

The influence of pheromones produced by freely growing laboratory populations on the reproductive maturation of prairie deermice following prenatal and postnatal exposure

Significantly more females reared on bedding soiled by different populations reproduced by 100 days of age than those reared on bedding soiled by the same populations (P<.1) or on clean bedding (P<.025) whereas the latter two groups did not differ significantly. Significantly greater percentages of females reproduced in both treatments maintained on soiled bedding than was true in the populations from which the bedding came. There were no significant differences between treatments in the weights of the reproductive organs (testes, seminal vesicles, ovaries, uteri) or in the numbers of corpora lutea or mature follicles in the ovaries of nulliparous females. The data suggest that if there were reproductively inhibiting pheromones influencing the population animals, they were altered or dissipated immediately after removal of the bedding from the population enclosures or that they were nonfunctional in the absence of the population animals. These data further suggest that the reproductive inhibition of population young is due to the presence of the populaton animals and that sociobiological factors within the populations act to prevent the reproductively stimulating influence of materials which may be present.     

Sexual incentive motivation, olfactory preference, and activation of the vomeronasal projection pathway by sexually relevant cues in non-copulating and naive male rats

There are some apparently healthy male rats that fail to mate after repeated testing with receptive females. We have previously shown that these "non-copulator (NC)" males show no partner preference for a receptive female when given the opportunity to physically interact with a sexually receptive female or a sexually active male. We also demonstrated that although NC males prefer odors from estrous females to odors from anestrous females, this preference is significantly reduced in comparison to the preference displayed by copulating (C) males. The aim of the present study was to evaluate in NC males sexual incentive motivation, that is, the approach behavior of male rats to either a sexually receptive female or a sexually active male in a test where the subjects can smell, hear, and see the stimulus animal but prevents their physical interaction. In addition, we determined whether NC rats have alterations in their ability to detect odors from conspecifics or odors related to food. In the detection of odors from conspecifics, we determined if these NC males are sexually attracted toward odors from receptive females or sexually active males. For food-related odors, we quantified the time it took the subjects to locate a hidden a piece of apple. Finally, using the induction of Fos-immunoreactivity (Fos-IR) as an index of neuronal activation, we compared the response of the vomeronasal projection pathway (VN pathway) of C and NC male rats exposed to estrous bedding. Males without sexual experience (WSE) were included in all experiments to determine the importance of previous heterosexual experience in the different behavioral tests and in the activity of the VN pathway. In the sexual incentive motivation test, we found that C and WSE male rats have a clear preference for estrous females over sexually active males, whereas NC male rats showed no preference. In odor tests, our results showed that C males had a clear preference for odors from estrous females as opposed to odors from sexually active males. Although NC and WSE male rats showed a preference for estrous female odors, this preference was significantly reduced compared to that shown by C males. No differences were found between WSE, C, and NC males in the detection of stimuli associated with food-related odors. A significant increase in Fos-IR was observed in the mitral cell layer of the accessory olfactory bulb in all groups when exposed to estrous bedding. However, only the C male rats exposed to estrous female bedding showed an increase Fos-IR in all structures of the VN pathway. An increase in Fos-IR was observed in the medial preoptic area (MPOA) of WSE males exposed to estrous bedding. No increases in Fos-IR were detected along the VN pathway in NC male rats. We proposed that NC male rats do not display sexual behavior due to a reduced sexual motivation that could be caused by alterations in the neuronal activity of the VN pathway during the processing of estrous odors.     

Antimutagenic effect of chemosignals from isolated female house mouse on male germ cells (Mus musculus L.)

The influence of chemosignals from isolated mature females of the CBA strain on level of spontaneous and radiation-induced meiotic disturbances in spermatocytes I of males of the same strain was studied. Using an ana-telophase method, 24-hour exposure of males to soiled bedding containing isolated females’ chemosignals was shown to lead to a significantly lower frequency of chromosomal aberrations and other meiotic disturbances in spermatocytes I as compared to males kept on clean bedding. The same effect of female chemosignals was found in the germ cells of irradiated males (4 Gr). The mechanisms and importance of the revealed antimutagenic effect of mouse female chemosignals on the male reproductive cells in the reproduction process are discussed.     

Social cues influence growth and sexual maturation of the male musk shrew (Suncus murinus)

Musk shrews were maintained from weaning (20 days of age) for 20 or 40 days in one of several social conditions. In Exp. 1, young males housed with adult females gained more weight and had heavier sex accessory organs than did young males housed with an adult male or reared alone. In Exp. 2 this same pattern of accelerated growth and sexual maturation was found when males were reared directly with an adult female or in a split cage where a wire barrier served to separate the male and his adult female cagemate. In Exp. 3, males were reared in cages containing clean or soiled bedding: soiled bedding was taken once every 5 days from the cage of an adult male, or a female. Under these conditions differences in the weights of reproductive tissues showed minimal variation with housing condition after 20 days of treatment. At that time males reared in soiled bedding taken from the cage of an adult female had accelerated development compared with control males. In Exp. 4, males were housed alone or in a split cage with an adult female which was separated by a wire mesh or a solid, opaque barrier. Males separated by a solid barrier from their female cagemates for 40 days had reproductive tissue weights equivalent to those measured in males reared alone. Taken together these results suggest that the presence of an adult female has dramatic effects on body growth and development of reproductive target tissues in young male musk shrews. Male-female social interactions could play an important role in the timing of puberty in this opportunistically breeding tropical mammal.     

Sexual Dimorphism of Mate Location in the Long-Toed Salamander Ambystoma macrodactylum columbianum

In breeding systems characterized by scramble competition among males, theory predicts that the efficient location of mating partners is more important to males than to females as a component of mating success. We experimentally tested in the laboratory the hypothesis that breeding male long-toed salamanders ( Ambystoma macrodactylum columbianum ), which scramble for mating opportunities, are better able to recognize and locate potential mates than are breeding females. Males were more likely to enter traps containing females than empty traps or traps containing males. Traps containing sponges soiled by females were more likely to attract males than traps containing clean (control) sponges, suggesting that chemical cues may be sufficient for mate location by males. Females were no more likely to enter traps containing males than empty traps. Our results are consistent with the theoretical prediction that selection has been stronger on male long-toed salamanders than on females in the context of capacity for recognizing and locating potential mating partners.     

注射胸腺依赖和非胸腺依赖抗原雄性黑线毛足鼠的内分泌状态和气味吸引力

作者研究了胸腺依赖抗原 (SRBC) (羊红血细胞 ,sheepredbloodcells ,SRBC)和非胸腺依赖抗原 (细菌脂多糖 ,lipopolysaccharide ,LPS)的免疫活化对黑线毛足鼠气味信号和内分泌状态的影响。成年雄鼠注射SRBS5天后 ,其气味的性吸引力下降 ,这种结果伴随着粪便中睾酮含量的下降。SRBC处理后 ,雄性气味吸引力降低 ,这在体液免疫反应低的雄性个体中最明显。与SRBC的作用相反 ,注射LPS的雄性个体的气味吸引力增加 ,成熟雌性个体用于嗅闻嗅觉刺激上的时间差异与经LPS和盐处理过的雄鼠的粪便中睾酮浓度的差异呈正相关。作者讨论了这两种相反的嗅觉效应的适应性意义     

Sexual reward in male rats: effects of sexual experience on conditioned place preferences associated with ejaculation and intromissions

Various behavioral models and studies have provided evidence suggesting that male rat sexual behavior has rewarding and reinforcing properties. However, there is little information regarding the rewarding values of the different components of sexual behavior. Therefore, this study used a conditioned place preference (CPP) paradigm to address whether ejaculation and intromissions differ in their rewarding incentive values. We also addressed whether the differential rewarding values were dependent on prior sexual experience. Sexually naïve and experienced males received one pairing of either intromissions or ejaculation with one of the chambers in the CPP box. The amount of time spent in each chamber of the CPP apparatus after conditioning was then measured. Both sexually naïve and sexually experienced males formed a CPP for ejaculation, while only sexually naïve, and not sexually experienced, males formed a CPP for intromissions. Moreover, in sexually naïve males, multiple pairings of ejaculation with the designated chamber resulted in a CPP relative to the control chamber paired with display of intromissions. These data support the hypothesis that there is a hierarchy of rewarding sexual behavior, with ejaculation being the most rewarding component, and that the rewarding incentive value of other components of sexual behavior is dependent upon prior sexual experience.