The effects of mating stimulation on c-Fos immunoreactivity in the female hamster medial amygdala are region and context dependent

During mating in hamsters, both tactile and nontactile sensory stimulation experienced by the female affect sexual behavior and progestational neuroendocrine reflexes. To test the interactions of these types of mating stimulation, c-Fos immunohistochemistry measured brain cellular activity during sexual behavior under conditions that included combinations of tactile and nontactile mating stimulation. Test groups received: (1) mating stimulation from a male, females being either fully mated or mated while wearing a vaginal mask, or (2) experimenter applied manual vaginocervical stimulation (VCS)-with or without males present, or (3) handling similar to VCS but without insertions-with or without males present. Numbers of c-Fos immunoreactive cells were counted in specific subdivisions of the posterior medial amygdala (MeP) and ventromedial hypothalamus (VMH). The medial amygdala dorsal and ventral subdivisions responded differentially to components of mating stimulation. The posterodorsal Me (MePD) cellular activation was greatest during mating conditions that included VCS and/or males present. However, the posteroventral Me (MePV) was sensitive to male exposure and not to VCS. Also, MePV and VMH shell responses mirrored each other, both being primarily sensitive to male exposure. In separate tests, manual VCS induced pseudopregnancy, though the procedure was most effective with additional nontactile stimulation from males present. In summary, contextual cues provided by nontactile male stimulation enhance the effect of vaginocervical and other tactile stimulation on reproductive processes. Furthermore, c-Fos expression in the female hamster medial amygdala is region and context dependent.     

Why do winners keep winning? Androgen mediation of winner but not loser effects in cichlid fish

Animal conflicts are influenced by social experience such that a previous winning experience increases the probability of winning the next agonistic interaction, whereas a previous losing experience has the opposite effect. Since androgens respond to social interactions, increasing in winners and decreasing in losers, we hypothesized that socially induced transient changes in androgen levels could be a causal mediator of winner/loser effects. To test this hypothesis, we staged fights between dyads of size-matched males of the Mozambique tilapia (Oreochromis mossambicus). After the first contest, winners were treated with the anti-androgen cyproterone acetate and losers were supplemented with 11-ketotestosterone. Two hours after the end of the first fight, two contests were staged simultaneously between the winner of the first fight and a naive male and between the loser of first fight and another naive male. The majority (88%) of control winners also won the second interaction, whereas the majority of control losers (87%) lost their second fight, thus confirming the presence of winner/loser effects in this species. As predicted, the success of anti-androgen-treated winners in the second fight decreased significantly to chance levels (44%), but the success of androgenized losers (19%) did not show a significant increase. In summary, the treatment with anti-androgen blocks the winner effect, whereas androgen administration fails to reverse the loser effect, suggesting an involvement of androgens on the winner but not on the loser effect.     

Mating increases male's interest in other females: a cognitive study in socially monogamous prairie voles (Microtus ochrogaster)

To determine whether socio-sexual interactions with females influence the male prairie vole's cognitive processing, three groups of males were simultaneously exposed to sensory stimuli of a control and a focal female then tested for their behavioral and neuronal responsiveness to the female cues. From the control female, all males received distal cues. From the focal female, the Unmated males received distal cues, the Unmated-Contact males received all cues but did not mate with her, and the Mated-Contact males received all cues and mated with her. Males were tested for their attentiveness to enclosures holding each female and for their memory of the females’ previous location. Males’ brains were then examined to localize activated regions following exposure to the odor of familiar versus unfamiliar focal females. The Mated-Contact males spent more time in the cage of the control female attending to her enclosure than in the cage of the focal female. Exposure to odors of unfamiliar focal females activated the cingulate cortex of Unmated-Contact males. There was a negative correlation between the level of neuronal activation in the retrosplenial cortex of males that were exposed to the odors of a familiar focal female and their attentiveness to the enclosure of the control female. The data suggest that the effect of socio-sexual interactions with a female on males’ cognition depends on the type of sensory signals males receive from females and how individual males perceive those signals.     

Experimental evidence that winning or losing a fight does not affect sperm quality in a field cricket

Fighting is a powerful social experience that can affect male reproductive behavior, including ejaculatory strategies. Whereas winners may monopolize females, losers may instead perceive high sperm competition and limited future mating opportunities, and accordingly enhance ejaculate quality to maximize their reproductive success. In male field crickets Gryllus bimaculatus that fight aggressively for control of breeding territories, winners are known to possess sperm of lower quality (viability) compared to losers, but it remains unclear whether this is due to short‐term fighting consequences. To test if the fighting experience per se (winning or losing) affects male adjustment of sperm viability, we subjected males to winning and losing experiences by staging fights against size‐matched rivals of known fighting ability. These rivals were males that previously won or lost a fight and, due to “winner‐loser effects” kept winning or losing subsequent contests. We sampled sperm prior and after the fight and twice in control males with no fighting experience and found no differences in sperm viability across measures. We conclude that males do not tailor their ejaculate quality following a single fight, or based on its outcome. Intrinsic differences in other attributes between winners and loser phenotypes may explain differences in sperm quality previously described in this system.     

Brain arginine vasotocin concentrations related to sexual behaviors and hydromineral balance in an amphibian

Arginine vasotocin (AVT), a potent stimulator of sexual behaviors and regulator of hydromineral balance in male rough-skinned newts (Taricha granulosa), was measured in 11 brain areas using microdissection and radioimmunoassay procedures. A 60-min test for sexual behaviors was used to segregate males into two groups: sexually responsive (initiated amplectic clasping behaviors) and sexually unresponsive (exhibited no sexual behaviors). Compared to sexually unresponsive males, sexually responsive males had significantly higher concentrations of immunoreactive (ir) AVT in the dorsal preoptic area, optic tectum, ventral infundibular nucleus, and cerebrospinal fluid. These results provide evidence for a behavioral action of endogenous AVT in T. granulosa. In another study, irAVT was measured in normal males (control newts maintained in water) and males that were dehydrated for 6 hr. Compared to normal males, dehydrated males had significantly lower concentrations of irAVT in the ventral preoptic area, but not in the other 10 areas of the brain. That different brain areas are associated with sexual behaviors and hydromineral balance suggests that there are some neuroanatomical separations between the behavioral and hydromineral aspects of the vasotocinergic system in this amphibian.     

Prenatal exposure to valproate induces sex-, age-, and tissue-dependent alterations of cholesterol metabolism: Potential implications on autism

Here, we investigated the protein network regulating cholesterol metabolism in the liver and brain of adolescent and adult male and female rats prenatally exposed to valproate (VPA), a well validated experimental model of autism spectrum disorders (ASD). We were aimed at studying whether prenatal VPA exposure affected the proteins involved in cholesterol homeostasis in a sex-dependent manner. To this aim the protein network of cholesterol metabolism, in term of synthesis and plasma membrane trafficking, was analyzed by western blot in the liver and different brain areas (amygdala, cerebellum, cortex, hippocampus, nucleus accumbens, and dorsal striatum) of adolescent and adult male and female rats prenatally exposed to VPA. Our results show that physiological sex-dependent differences are present both in the liver and in brain of rats. Interestingly, VPA affects specifically the brain in an age- and region-specific manner; indeed, cerebellum, cortex, hippocampus and nucleus accumbens are affected in a sex-dependent way, while this does not occur in amygdala and dorsal striatum. Overall, we demonstrate that each brain area responds differently to the same external stimulus and males and females respond in a different way, suggesting that this could be related to the diverse incidences, between the sexes, of some neurodevelopmental pathologies such as autism, which displays a 3:1 male to female ratio.     

Effect of mutation-induced excess brain concentration of intermediates of the kynurenine pathway of tryptophan metabolism on stress resistance and courtship behavior and communicative sound production in male Drosophila melanogaster

The resistance of courtship behavior and communicative sound production to heat shock (37 degrees C, 30 min) was studied in wild-type Canton S (CS) male Drosophila melanogaster and males of two strains with defects in the kynurenine pathway of tryptophan metabolism (KPTM) caused by mutations cinnabar (block at the level of kynurenine-3-hydroxylase leading to accumulation of kynurenic acid, a neuroprotective metabolite, in the brain) and cardinal (block at the level of phenoxazinone synthetase causing accumulation of 3-hydroxykynurenine, an oxidative stress generator, in the brain). Males of each strain were divided into four groups. Males from control groups were not exposed to heat shock. The other groups were exposed to heat shock at the late embryonic/early larval (the first instar) developmental stage, when mushroom bodies are formed (HS1 groups); at the prepupal stage, when the brain central complex develops (HS2 groups); or at the imago stage 1 h before the experiment (HS groups). All males were tested at an age of five days. Virgin and fertilized five-day-old CS females served as courtship objects. The courtship behavior and singing of control CS and cinnabar males were similar. Control cardinal males also had high motivation, but their courtship efficiency was lower because of less precise movements (wing vibration was often not accompanied by sound production) and hyperexcitability. Exposure of first-instar larvae to heat shock had almost no effect on behavior or singing of adult males of any strain. In cardinal males exposed to heat shock at the prepupal stage or, especially, at the imago stage 1 h before the test (the HS2 and HS groups), courtship was strongly impaired, and various distortions appeared in their sound signals, which indicated disturbance of coordination between elements of the song center and their interaction with pacemakers. These effects were much milder or absent altogether in HS2 and HS wild-type males and, especially, cinnabar males. Thus, permanent excess of 3-hydroxykynurenine in the male brain dramatically decreased their stress resistance. In contrast, excess of kynurenic acid alleviated the consequences of stress.     

Is full physical contact necessary for buffering effects of pair housing on social stress in rats?

Our previous study showed that pair housing with a familiar male prevented an increase in anxiety caused by social defeat in male rats. The present study attempted to identify the aspects of social interactions with a familiar male that are needed for the emergence of such a pair-housing effect. In Experiment 1, the subject was repeatedly exposed to the cage and bedding used by a familiar pairmate, after two instances of social defeat. Mere exposure to the soiled cage and bedding did not prevent an increase in anxiety in the elevated plus-maze test performed two weeks after social defeat. In Experiment 2, the subject was separated from a familiar pairmate with a wire mesh partition, which allowed visual, auditory, and limited physical contact, in addition to olfactory contact with the pairmate. The separation with a wire mesh partition abolished the buffering effect of pair housing on anxiety. These results indicate that visual, auditory, and olfactory contact with a familiar male was not sufficient in reducing the anxiogenic effect of social defeat in male rats. It was suggested that full physical contact is necessary for the emergence of the buffering effect of pair housing on social stress.     

Dominant-subordinate relationships in hamsters: sex differences in reactions to familiar opponents

In the majority of mammalian species, males are dominant over and more aggressive than females. In contrast, some reports suggest that female golden hamsters are more aggressive than males but systematic comparisons using the same methods for both sexes are rare. We observed same-sexed pairs of hamsters over repeated trials to assess whether sex differences existed in the level of agonistic behavior and in the development and maintenance of dominant-subordinate relationships with familiar partners. There were no sex differences in measures of agonistic behavior or fear responses (fleeing) during the initial series of three trials on the first day of testing. Following a four-day interval, males that had lost in session 1 showed fearful responses to a familiar dominant male and were not likely to engage in a fight with him. In contrast, females that lost the initial fights were not fearful and fought vigorously with the familiar winner in subsequent encounters. Although the amount of agonistic behavior engaged in by females did decrease over the course of the three sessions, females that lost did not demonstrate an increase in fear, as measured by the latency to flee. Males that lost fights did show increased fear during later trials and sessions. These results suggest that female hamsters are less affected by losing fights than males are and thus that females are less likely than males to develop highly polarized dominant-subordinate relationships. Further work is needed to understand the mechanisms underlying these sex differences.     

Determinants of Dominance in the Tree Lizard Urosaurus ornatus: the Relative Importance of Mass,Previous Experience and Coloration

A determination of some of the factors that predict the outcome of contests between male tree lizards, Urosaurus ornatus, was made using logistic regression modelling on matched-pair data. Two-day-long encounters were staged between pairs of males differing in size (snout-vent length and mass), previous contest status (previous winners and previous losers), and coloration (dorsal coloration during their previous contest and throat coloration, a fixed trait). Mass proved to be the best single predictor of contest outcome, resulting in an 80% correct classification rate for predicting winners and losers, far better than the less than 57% correct classification rate for snout-vent length. Previous social status (winner or loser) also was a powerful single predictor of contest outcome with a 793% correct classification rate, as was previous dorsal coloration (76.7%). When combined, mass and previous status produced the strongest combination of predictors with a better than 86% correct classification rate. Contrary to several previous studies, which implicated throat coloration as an important status signal of dominance, our results failed to show that throat coloration is a strong predictor of contest outcome. Possible reasons for this discrepancy with earlier findings are discussed. The logistic regression models also allow prediction of the magnitude of difference in mass between two contestants for there to be an equal chance of winning, given a second asymmetry in contest predictors.     

Effect of mutation-induced excess brain concentration of intermediates of the kynurenine pathway of tryptophan metabolism on stress resistance of courtship behavior and communicative sound production in male Drosophila melanogaster

The resistance of courtship behavior and communicative sound production to heat shock (37°C, 30 min) was studied in wild-type Canton S (CS) male Drosophila melanogaster and males of two strains with defects in the kynurenine pathway of tryptophan metabolism (KPTM) caused by mutations cinnabar (block at the level of kynurenine-3-hydroxylase leading to accumulation of kynurenic acid, a neuroprotective metabolite, in the brain) and cardinal (block at the level of phenoxazinone synthetase causing accumulation of 3-hydroxykynurenine, an oxidative stress generator, in the brain). Males of each strain were divided into four groups. Males from control groups were not exposed to heat shock. The other groups were exposed to heat shock at the late embryonic/early larval (the first instar) developmental stage, when mushroom bodies are formed (HS1 groups); at the prepupal stage, when the brain central complex develops (HS2 groups); or at the imago stage 1 h before the experiment (HS groups). All males were tested at an age of five days. Virgin and fertilized five-day-old CS females served as courtship objects. The courtship behavior and singing of control CS and cinnabar males were similar. Control cardinal males also had high motivation, but their courtship efficiency was lower because of less precise movements (wing vibration was often not accompanied by sound production) and hyperexcitability. Exposure of first-instar larvae to heat shock had almost no effect on behavior or singing of adult males of any strain. In cardinal males exposed to heat shock at the prepupal stage or, especially, at the imago stage 1 h before the test (the HS2 and HS groups), courtship was strongly impaired, and various distortions appeared in their sound signals, which indicated disturbance of coordination between elements of the song center and their interaction with pacemakers. These effects were much milder or absent altogether in HS2 and HS wild-type males and, especially, cinnabar males. Thus, permanent excess of 3-hydroxykynurenine in the male brain dramatically decreased their stress resistance. In contrast, excess of kynurenic acid alleviated the consequences of stress.     

Isotocin regulates paternal care in a monogamous cichlid fish

While the survival value of paternal care is well understood, little is known about its physiological basis. Here we investigate the neuroendocrine contributions to paternal care in the monogamous cichlid, Amatitlania nigrofasciata. We first explored the dynamic range of paternal care in three experimental groups: biparental males (control fathers housed with their mate), single fathers (mate removed), or lone males (mate and offspring removed). We found that control males gradually increase paternal care over time, whereas single fathers increased care immediately after mate removal. Males with offspring present had lower levels of circulating 11-ketotestosterone (11-KT) yet still maintained aggressive displays toward brood predators. To determine what brain regions may contribute to paternal care, we quantified induction of the immediate early gene c-Fos, and found that single fathers have more c-Fos induction in the forebrain area Vv (putative lateral septum homologue), but not in the central pallium (area Dc). While overall preoptic area c-Fos induction was similar between groups, we found that parvocellular preoptic isotocin (IST) neurons in single fathers showed increased c-Fos induction, suggesting IST may facilitate the increase of paternal care after mate removal. To functionally test the role of IST in regulating paternal care, we treated biparental males with an IST receptor antagonist, which blocked paternal care. Our results indicate that isotocin plays a significant role in promoting paternal care, and more broadly suggest that the convergent evolution of paternal care across vertebrates may have recruited similar neuroendocrine mechanisms.     

Resting cortisol levels and the emergence of dominant status among male vervet monkeys

Resting serum cortisol was measured in adult male vervet monkeys (Cercopithicus aethiops sabaeus) in four different conditions: (1) among groups with unaltered group membership and established dominance hierarchies; (2) among groups from which the original dominant male had been removed and in which the remaining males competed for dominant status; (3) among newly formed groups of three unfamiliar males each of which had been the dominant male in his previous group; and (4) among groups from which a dominant male was temporarily separated and returned. In Condition 1, cortisol concentrations did not differ between dominant and subordinate males. The second condition showed that cortisol levels were highest among males who eventually emerged as the dominant male. In the third condition, however, cortisol levels did not differentiate eventually dominant from eventually subordinate males. In the last condition, cortisol levels were highest in the animals that became or remained dominant following reintroduction. These data indicate that cortisol concentration does not differ between dominant and subordinate males in stable groups and that cortisol rises during competition for dominance among familiar males.     

Integration of New Males into Four Social Groups of Tufted Capuchins (Cebus apella)

We examined how aggressive, affiliative, and sexual behavior function to integrate male capuchins (Cebus apella) into a new social group. Nine males were exchanged among four social groups. We performed instantaneous scans and all-occurrence sampling during baseline, introduction, and follow-up periods. The study included three different introduction situations: 1) males familiar to one another were introduced to a group with no other adult male, 2) males unfamiliar to one another were introduced to a group with no other adult male, and 3) males familiar to one another were introduced to a group with an existing elderly, resident male. Severe aggression occurred in situations 2 and 3, but the introductions were peaceful in situation 1. In all cases proceptive females were among the first individuals to affiliate with the males, and males did not appear to compete for access to proceptive females. Following their period of proceptivity, the females that had cycled remained preferred social partners for the males. Immature animals also quickly affiliated with the new males, and the males tolerated the attention from immatures. Affiliative relationships between the males and nonproceptive females developed slowly, and while male-female aggression was mild, aggression among adult males (familiar and unfamiliar) had the potential to be severe.     

Fighting dynamics of male copperheads,Agkistrodon contortrix (Serpentes,Viperidae): Stress-induced inhibition of sexual behavior in losers

Adult male copperheads (Agkistrodon contortrix) fight for priority of access to females during the mating periods in spring and late summer. During fights, one male abruptly quits and retreats, and the other chases in pursuit. One male thus emerges as the winner and the other as the loser. Reversal of this outcome does not occur during the time of observation (30 min) nor in 24 hr postfight trials. In all cases, winners gain priority of access to females. Losers, in contrast, do not pursue females nor gain access to them, even when winners are removed from the arena. In this study, courtship performance of male A. contortrix was studied in the laboratory using subjects with either winning or losing experience from staged fights. All males used first were tested with a single female to determine courtship performance prior to the agonistic trials. From these tests, each male was given a single courtship score of 0 (no courtship) to 3 (most intense courtship). Only males receiving a score of 2–3 were used in the initial agonistic trials. All staged fights were conducted in a large arena and involved two males and one female. Following fights, winners and losers were tested again for courtship performance. In trials conducted at 24 hr and 7 days postfight, only losers were tested. It was found that prefight courtship scores were not significantly different between winners and losers. At 30 min postfight, most losers showed complete suppression of courtship behavior (score 0). Winners, in contrast, showed equivalent or an increase in their courtship scores. At 24 hr postfight, courtship scores of losers remained significantly lower than their prefight scores. At 7 days postfight, courtship scores of losers were not significantly difference from their original prefight scores. Fighting behavior in free-ranging A. contortrix represents a potentially significant cost to losers if it is associated with loss of reproductive opportunities. Recent evidence from hormonal studies indicates that inhibition of courtship and fighting behavior in male A. contortrix is stress-induced. © 1996 Wiley-Liss, Inc.     

Male Competition in a Wandering Spider (Cupiennius getazi,Ctenidae)

We describe male-male competition in a wandering spider living on plants (Cupiennius getazi, Ctenidae) and discuss it within the general context of the mating system. 1. Males produce vibratory courtship signals (duration about 20 s) and competition signals (2 s). Upon exposure to female silk, males produce almost exclusively courtship signals (98%) if alone or in the presence of a female. In the presence of a rival alone, an average of 25% of a male's vibratory signals are courtship signals and 75% competition signals. In the presence of both a rival and a female, an average of 50% are courtship and 50% competition signals. Females respond to both male courtship and/or competition signals with vibratory courtship whereas males react by vibratory competition. The intensity of the reaction of both males and females is independent of the signal type. 2. Males displaying vibratory signals move slowly over the plant and repel attacks from rivals and females with extended front legs. Pairs of males interact in three ways. (i) Both males produce vibratory signals; one of them leaves the plant (53% of 90 trials). (ii) Both males vibrate, approach and touch (20%) or pounce on each other (20%). (iii) A male approaches the signalling opponent without producing vibrations and attacks him (7%). This is a conditional vibrocryptic tactic. The presence of a female incites male competition. Males do not interact with the female but approach each other (in 24% of the 26 trials “vibrocryptically”) and escalate more often (88%) and more quickly to overt fight than in the absence of a female. The male remaining on the plant approaches the female. 3. Male-male fights are ritualized. During 64 bodily contacts no male was injured. Males exposed to female silk and males using the vibrocryptic tactic were more often the winners of an interaction than males not exposed to female silk and than males vibrating while approaching their rival. The outcome of fights is not correlated with age, leg length, body weight and rate of signalling when no female is present. In contrast, body weight and leg length determine the outcome when a responding female is present, the larger male being the winner. 4. Intrasexual and intersexual interactions suggest that both male competition and female choice mechanisms may regulate sexual selection in this species.     

Chronic social stress in puberty alters appetitive male sexual behavior and neural metabolic activity

Repeated social subjugation in early puberty lowers testosterone levels. We used hamsters to investigate the effects of social subjugation on male sexual behavior and metabolic activity within neural systems controlling social and motivational behaviors. Subjugated animals were exposed daily to aggressive adult males in early puberty for postnatal days 28 to 42, while control animals were placed in empty clean cages. On postnatal day 45, they were tested for male sexual behavior in the presence of receptive female. Alternatively, they were tested for mate choice after placement at the base of a Y-maze containing a sexually receptive female in one tip of the maze and an ovariectomized one on the other. Social subjugation did not affect the capacity to mate with receptive females. Although control animals were fast to approach females and preferred ovariectomized individuals, subjugated animals stayed away from them and showed no preference. Cytochrome oxidase activity was reduced within the preoptic area and ventral tegmental area in subjugated hamsters. In addition, the correlation of metabolic activity of these areas with the bed nucleus of the stria terminalis and anterior parietal cortex changed significantly from positive in controls to negative in subjugated animals. These data show that at mid-puberty, while male hamsters are capable of mating, their appetitive sexual behavior is not fully mature and this aspect of male sexual behavior is responsive to social subjugation. Furthermore, metabolic activity and coordination of activity in brain areas related to sexual behavior and motivation were altered by social subjugation.     

Friend or foe: behavioral responses to conspecifics in the northern scorpion, <Emphasis Type="Italic">Paruroctonus boreus</Emphasis> (Scorpionida: Vaejovidae)

We investigated the behavioral response of male Paruroctonus boreus to male and female pheromones of conspecifics. Adult male scorpions actively search for females during the mating season, but lack visual cues to direct their movement. Chemical cues (pheromones) are thought to be responsible for directing movement. In a series of laboratory experiments, in Y-mazes, males were exposed to areas that had contained males and females from the same population and areas that had contained conspecifics from different populations. Males spent significantly more time in areas that had contained females from the same population but did not show a preference for areas exposed to females from different populations. Males also significantly avoided areas exposed to males from the same population. Our results suggest that P. boreus has a sex-specific response to pheromones and that this response may be population-specific.     

Stimuli from conspecifics influence brain mast cell population in male rats

It is well established that mast cells occur within the brain of many species, and that the brain mast cell population is not static, but changes with the behavioral and physiological state of the animal. In this study, we tested whether exposure to conspecifics alters the number of brain mast cells in male rats, and then investigated the nature of stimuli influencing the changes observed in the number and localization of brain mast cells. Five days of cohabitation with an ovariectomized, estrogen-progesterone (OVX + EP)-treated female resulted in the largest number of thalamic mast cells, while pairing with such a female physically separated by a wire mesh or with a novel male produced a smaller, but significant increase over other pairings (OVX females for 5 days, OVX and OVX + EP females for 1 day, familiar or isolated males for 5 days). In all groups, mast cells were localized within specific dorsal thalamic nuclei, including the paraventricular nucleus, anterior nuclear group, or mediodorsal, ventroposterior, or medial geniculate nuclei. The results suggest that the behavioral and/or endocrine factors associated with cohabitation with conspecifics are sufficient to alter the number of brain mast cell-specific nuclei in the thalami of male rats and thus can provide targeted delivery of neuromodulators to specific regions of the brain that process information concerning the normal physiological state of the animal.