Male Copulation Research Articles

Dalliances and doubtful dads: what determines extra-pair paternity in socially monogamous wandering albatrosses?

Genetic techniques have revealed surprisingly high rates of extra-pair paternity (EPP) in socially monogamous albatrosses. We sought to establish social and genetic influences on EPP in wandering albatrosses (Diomedea exulans) at Marion Island, where EPP rates were 14–24 % in three successive seasons. EPP probably resulted from both female solicited extra-pair behaviours and male forced copulations. EPP was not linked to breeding experience nor with poor reproductive performance, despite a tendency for pairs to consistently produce either EPP or within-pair paternity (WPP) chicks. Mate guarding may inhibit extra-pair behaviour; however, parental arrival date and presence in the colony prior to laying did not correlate with EPP. There was little support for genetic advantages to producing EPP chicks, but the population is characterised by low genetic variability, which may result in mate incompatibility. Mates of pairs that failed and pairs producing EPP young tended to be more similar genetically to their partners than mates producing WPP young, suggesting that EPP may counter mate incompatibility. EPP and WPP chicks grow equally well, so cuckolded males did not reduce investment in EPP chicks. The lack of discriminatory behaviour by cuckolded males together with low genetic diversity in the population may allow continued high levels of EPP. In albatrosses, pair bonds are typically long lasting and the costs of forming new pairings may discourage mate swapping. Females may undertake extra-pair copulations as an adaptive alternative to mate swapping because the costs of extra-pair behaviour are small.

Characterization of the Spinal Nucleus of the Bulbocavernosus Neuromuscular System in Male Mice Lacking Androgen Receptor in the Nervous System

Motoneurons in the spinal nucleus of the bulbocavernosus (SNB) and their target bulbocavernosus (BC) and levator ani (LA) muscles play a role in male copulation and fertility. Testosterone (T) induces sexual differentiation of this SNB neuromuscular system during development and maintains its activation in adulthood. In the rat, T-induced effects mostly involve the androgen receptor (AR). However, the role of central AR in T-induced effects remains to be studied with pertinent genetic models. We addressed this question by using specific motoneuron immunolabeling and retrograde tracing in mice selectively disrupted for AR in the nervous system. This work reveals that nervous system AR is not required either for T-induced development of BC-LA muscles and perinatal sparing of SNB motoneurons from atrophy or for adult sensitivity of BC-LA muscles to T. By contrast, loss of AR expression in the nervous system resulted in SNB motoneurons having smaller somata and shorter dendrites than controls. We studied the effects of adult castration and T supplementation on SNB cell morphology in control and mutant males; these experiments showed that central AR is involved in the developmental regulation of soma size and dendritic length and in the adult maintenance of soma size of SNB motoneurons. T seemed to act indirectly through BC-LA muscles to maintain dendritic length in adulthood. Our results also suggest that central AR functions may contribute to normal activity of SNB motoneurons and perineal muscles because mutant mice displayed diminished copulatory behavior and fertility.

Sexual responses of the male rat medial preoptic area and medial amygdala to estrogen I: Site specific suppression of estrogen receptor alpha

Male rat copulation is mediated by estrogen-sensitive neurons in the medial preoptic area (MPO) and medial amygdala (MEA); however, the mechanisms through which estradiol (E2) acts are not fully understood. We hypothesized that E2 acts through estrogen receptor α (ERα) in the MPO and MEA to promote male mating behavior. Antisense oligodeoxynucleotides (AS-ODN) complementary to ERα mRNA were bilaterally infused via minipumps into either brain area to block the synthesis of ERα, which we predicted would reduce mating. Western blot analysis and immunocytochemistry revealed a knockdown of ERα expression in each brain region; however, compared to saline controls, males receiving AS-ODN to the MPO showed significant reductions in all components of mating, whereas males receiving AS-ODN to the MEA continued to mate normally. These results suggest that E2 acts differently in these brain regions to promote the expression of male rat sexual behavior and that ERα in the MPO, but not in the MEA, promotes mating.

THE PRECOPULATORY FUNCTION OF MALE GENITAL SPINES IN DROSOPHILA ANANASSAE[DOLESCHALL] (DIPTERA: DROSOPHILIDAE) REVEALED BY LASER SURGERY

That male genital morphology evolves via postcopulatory sexual selection is a widely held view. In contrast, the precopulatory sexual selection hypothesis for genital evolution has received less attention. Here, we test the hypothesis that male genital spines of Drosophila ananassae promote competitive male copulation success. Using laser surgery to manipulate trait size, we demonstrate that incremental reductions of spine length progressively reduce male copulation success: males without spines failed entirely to copulate because of an inability to couple the genitalia together, whereas males with halfway ablated and blunted spines suffered reductions in copulation success of 87% and 13%, respectively. The decrease in copulation success resulting from spine length reduction was markedly stronger in sexually competitive environments than in noncompetitive environments, and females expressed resistance behaviors similarly toward competing male treatments, demonstrating directly the role of genital spines in promoting competitive copulation success. Because these spines are widespread within Drosophila, and because genital traits with precopulatory function are being discovered in a growing number of animal taxa, precopulatory sexual selection may have a more pervasive role in genital evolution than previously recognized.

Fluoxetine Chronic Treatment Inhibits Male Rat Sexual Behavior by Affecting Both Copulatory Behavior and the Genital Motor Pattern of Ejaculation

Selective serotonin reuptake inhibitors (SSRIs) are antidepressants that cause sexual dysfunction. The SSRI fluoxetine (FLX) appears to particularly affect the ejaculatory response. Ejaculation is regulated both at brain and spinal levels. To study the acute and chronic effects of FLX on male rat copulatory behavior, trying to distinguish between brain and spinal cord FLX-induced changes on the ejaculatory response. Sexually experienced male rats were intraperitoneally injected with 5 or 10 mg/kg FLX and tested for sexual behavior during 60 minutes on days 1, 7, and 14 of treatment. After a 2-day drug holiday, the males chronically treated with the high FLX dose were spinalized to record spontaneous and mechanically evoked genital motor patterns of ejaculation (GMPEs). In addition, independent groups were used to evaluate the acute effects of 1, 3, or 10 µg/rat FLX (intravenously) on the GMPE. Number of ejaculatory series and their parameters; electromyographic recordings of the GMPE in the bulbospongiosus muscles and their parameters. Acute FLX injection slightly affected sexual behavior display and dose-dependently inhibited the expression of the GMPE. Chronic FLX treatment did not inhibit copulation but produced deficits in the parameters related to ejaculation after the high dose. In these animals, the response capacity of the spinal generator of ejaculation (SGE) as well as the number of discharges in the GMPE was decreased as a result of chronic FLX treatment. Chronic FLX treatment produces inhibitory effects on male rat copulation, particularly on ejaculation, some of which are exerted directly at the SGE.

A female‐produced short‐range sex pheromone in the egg parasitoid Trissolcus brochymenae

AbstractThe potential for short‐range sex pheromone communication by the egg parasitoid wasp Trissolcus brochymenae (Hymenoptera: Platygastridae) was investigated in closed arena bioassays. Males of this parasitoid showed more antennal drumming and more frequent mounting behaviour on 1‐ to 2‐d‐old virgin females compared with 8‐d‐old virgin females. Male copulation attempts were fewer with previously mated females than with virgin females. Males courted and made copulation attempts with 1‐ to 2‐d‐old female cadavers, but not with male cadavers or with female cadavers rinsed in organic solvents of different polarities. Male attraction to female cadavers was re‐established by treating cadavers with acetone extracts of females, but not with ether or hexane extracts. In experiments using female cadavers dissected into head, mesosoma, and gaster, and then reassembled using one unwashed body section and two body sections washed in acetone, males were attracted only to the reassembled cadavers with an unwashed mesosoma. These findings suggest that (1) courtship behaviour in males of T. brochymenae is triggered by a short‐range sex pheromone produced by females; (2) the age and the physiological condition of females (virgin/mated) influence pheromone release or production; (3) the female's mesosoma is the source of the sex pheromone; and (4) polar components of the sex pheromone play a major role in influencing male behaviour. Our results suggest that quasi‐gregarious egg parasitoids are selected for short‐range rather than long‐range sex pheromones.

Male preference and female cues: males assess female sexual maturity and mating status in a web-building spider

Reproductive success relies on communication signals used by females to exert mate attraction and assessment of female reproductive value by males. During mate search males of the spider, Stegodyphus lineatus experience high mortality and low female encounter rate. Females vary in sexual maturity and mating status (virgin and mated) and hence in reproductive value for males, which should influence male mating effort. Due to high costs of remating, only virgin females are expected to signal mate attraction. Males would benefit from mating preferentially with virgins due to the costs of overcoming mated females' resistance, although they gain paternity share. Males should avoid immature females, unless guarding precopulatory to access females on maturation. Low encounter rates predict males to invest also in females of inferior reproductive value. We investigated male ability to discriminate immature, virgin, and mated females by assessing: the number of females males visit in the field; male mating effort and male discrimination of females based on silk cues in laboratory trials. In the field, males were found most frequently with virgin females. Male mating effort, copulation success, and preference based on silk cues were higher with mature compared with immature females. Our data suggest that females signal sexual receptivity and that males are able to discriminate both sexual maturity and mating status, however, males do not exert strong preference for virgins. The combination of high costs of mate search and low encounter rate likely exerts selection on males to mate indiscriminately and overcome female resistance to remating.

Sneaky Monkeys: An Audience Effect of Male Rhesus Macaques (Macaca mulatta) on Sexual Behavior

Males and females have different sexual interests and subsequently may show conflicting sexual strategies. While dominant males try to monopolize females, promiscuity benefits females and subordinate males. One way to escape monopolization by dominant males is to copulate in their absence. We tested this inhibitory effect of males on the sexual behavior of their group members in captive group-living Rhesus macaques. Copulations between females and nonalpha males almost exclusively took place when the alpha male was out of sight. Furthermore, the inhibiting effect was not unique for the alpha male. An upcoming nonalpha male also inhibited copulations of its group members, and three other nonalpha males inhibited female copulation solicitations. Females adjusted their behavior to the presence of bystander males, as they initiated and accepted initiations more often in absence than in presence of bystander males. Although not significant, in males, a similar pattern was found. The observed reduction in mating behavior in presence of bystander males is in accordance with an "audience effect," in which the behavior is modulated in relation to the presence or absence of third parties. This audience effect may serve as an important mechanism to reduce (aggressive) interruptions of subordinate male copulations.

Drosophila female precopulatory behavior is modulated by ecdysteroids

The effect of ecdysteroid signaling on Drosophila female precopulatory behavior was investigated using two types of mutants with either globally reduced ecdysteroid availability or reduced expression of ecdysone receptors in fruitless neurons, known to control sexual behavior. While being courted by males, mutant females performed significantly less full ovipositor extrusion behavior to reject male copulation attempts. Ecdysteroid depleted females (ecdysoneless1) performed male-like courtship behaviors, including unilateral wing extension and song production with patterns very similar to male courtship song. These results support the hypothesis that ecdysteroids modulate female sexual behavior, perhaps acting as a regulator of sexual motivation, and as a component affecting the performance of sex specific behavior patterns.

Fruitless RNAi knockdown in the desert locust, Schistocerca gregaria, influences male fertility

In Drosophila melanogaster, the male-specific splice isoform of the fruitless gene (FruM) encodes a set of transcription factors that are involved in the regulation of male courtship and copulation. Recent insights from non-drosophilid insects suggest a conserved evolutionary role for the transcription factor Fruitless. In the desert locust, Schistocerca gregaria and the German cockroach, Blatella germanica, both orthopteran insects, a conserved functional role for fruitless has been proposed. Fru specific RNAi knockdown in the third nymphal stage of male Schistocera gregaria delays copulation initiation and results in reduced progeny. In order to identify the origin of the observed phenotypic effects following a fruitless RNAi treatment in the male, we show that the fru knockdown has no detectable effect on spermio- or spermatogenesis and on the transfer of spermatozoa during copulation. Nevertheless, it is clear that the male seminal vesicles contain significantly less spermatozoa after fru RNAi as compared to gfp RNAi controls. We conclude that a lowered male fertility, caused by the fru knockdown in male desert locusts may be the direct cause for the reduction of the progeny numbers in their naïve female copulation partners.

Male reproductive investment relative to age and flight behaviour in the monandrous butterfly Pararge aegeria

Male reproductive investment may signify a considerable cost to male insects that produce sperm packages or spermatophores. Male butterflies allocate much of their active time budget to mate location, and they may adopt different behavioural strategies to do so. In the speckled wood butterfly (Pararge aegeria L.), males adopt either a territorial wait-and-fight strategy (territorial perching) or a fly-and-search strategy in wider areas (patrolling). In this study, we analysed the impact of male age, male size and male behaviour (i.e. behavioural strategies and levels of activity) on spermatophore investment (i.e. spermatophore mass, number of eupyrene sperm bundles). As predicted, reproductive investment increased with male age and size. Nevertheless, the increase of spermatophore mass with age and the number of eupyrene sperm bundles (i.e. fertile sperm) was stronger in low-activity males compared to active flying males. This suggests that flight activity has a negative impact on male reproductive investment. However, males that were forced to fly in the laboratory produced more eupyrene sperm bundles than resting males. We discuss the potential effects of male–male competition and predation risk on current versus future male reproduction. Males adopting different mate-locating strategies (perching and patrolling) in outdoor cages did not differ in spermatophore traits as was predicted from their very different flight performances. Copulations of territorial perching males took somewhat longer than copulations with non-perching males. There was a significant family effect of spermatophore size and of the expression of male mate-locating strategies suggesting heritable variation. Female traits (i.e. age and size) did not strongly affect spermatophore production. We discuss the results relative to both ultimate and proximate explanations of the complex relationships between butterfly activity, behavioural strategies, age and spermatophore production.

The protein synthesis inhibitor anisomycin reduces sex behavior during a critical period after testosterone treatment in male Syrian hamsters

Testosterone (T) is critical for maintaining male sexual behavior (MSB) in rodents, in part by altering protein synthesis in a well-defined neural circuit. The specific timing of protein synthesis essential for expression of MSB has never been investigated. We administered the protein synthesis inhibitor anisomycin (Ani) to castrated male Syrian hamsters treated sc with 100μg T in an aqueous vehicle once weekly; this T regimen maintains MSB while elevating circulating T concentrations for only a few hours after each injection. Hamsters were injected sc with the vehicle or 12.5mg Ani at one of several times relative to T administration; MSB was assessed once per week, 6days after the previous T injection, for 5weeks. Anisomycin administered 6–12h after T injection significantly reduced the expression of sexual behavior, whereas Ani treatment between 3h before and 3h after T injection did not impair MSB. This experiment is the first to assess the specific timing of protein synthesis relative to a T pulse that is required for the expression of MSB. The demarcation of a critical interval for T-induced protein synthesis necessary for maintenance of MSB should facilitate specification of the genomic, proteomic, and biochemical cascades that subserve actions of T on male copulation.

New information on the evolution of mating behaviour in Sepsidae (Diptera) and the cost of male copulations in Saltella sphondylii

Here we describe the hitherto unknown details of the highly unusual mating behaviour of Saltella sphondylii—a widely cited model for male longevity costs caused by multiple copulations. When compared to the known mating behaviour of 28 sepsid species, we find five unique behavioural elements based on frame-by-frame analyses of video-recordings. These new behaviours are documented with video clips. We suggest that the male longevity costs could be due to copulation bouts that involve multiple insertions of a comparatively membranous phallus into the female. We compare the phallus of the Saltella sphondylii to those from three other species (Themira putris, Parapaleosepsis plebeia, Sepsis punctum).

Light conditions affect sexual performance in a lekking tephritid fruit fly

Sensory systems are very susceptible to early environment experience. Mating success depends on the transmission of information from the signaller to the receiver, which means that sensory biases caused by developmental environment are likely to affect sexual selection. We investigated the impact of the developmental visual environment (light spectrum) on male copulation behaviour and female preference in the lekking tephritid Anastrepha ludens. We reared flies in four different light spectrum conditions - red light, blue light, shaded light and darkness - during their first 16 days after emerging from pupae. We found that the light environment experienced during early adulthood affected mating frequency and, in some cases, the latency to copulate, but not copulation duration. Males exposed to any of the three light treatments (red, blue or shaded light) were more frequently chosen as mating partners than dark-reared males. Flies reared under dark conditions exhibited the lowest mating performance out of any of the rearing environments. Under field cage conditions, a slight assortative mating between blue- and red-light-reared flies was detected. Additionally, females reared in blue light and darkness mated less compared with females reared in red and shaded light. Our data demonstrate that male mating behaviour is flexible in response to light environment. The findings suggest that light spectrum only weakly affects the direction of sexual selection by female choice; however, dark rearing environments deeply affect mating success.

Ginkgo biloba treatment increases copulation but not nNOS activity in the medial preoptic area in male rats

Nitric oxide (NO) is an important messenger in the central nervous system to mediate male copulatory behavior. EGb 761, a standardized extract of Gingko biloba, has been reported to facilitate male copulation in rats. The present study is to determine the effects of neuronal nitric oxide synthase (nNOS) in the medial preoptic area (MPOA) on copulation in male rats following EGb 761 treatment. Adult male rats were treated with 50 mg/kg of EGb 761 or distilled water by oral gavage for 14 consecutive days. The animals were sacrificed approximately 14 h after the last behavioral test and MPOA brain tissues were collected for nNOS immunohistochemistry. EGb 761 treatment for 14 days significantly increased the intromission frequency compared to the vehicle-treated controls on day 14. An increase in ejaculation frequency was also seen in the EGb 761-treated group compared to the vehicle-treated controls on day 14 and to the same group on day 0. However, EGb 761 treatment did not influence the number of nNOS-immunoreactive cells in the MPOA. These results suggest that enhanced male copulatory performance in sexually experienced rats administered EGb 761 may not be related to central nNOS activity in the MPOA.

Carry-over effects of male extra-pair copulation opportunity on biparental effort in zebra finches

Whether parental effort can be negotiated between partners over ecological time and adjusted across different contexts is not well understood. We manipulated male extra-pair copulation (EPC) opportunity in captive zebra finches, Taeniopygia guttata, to test whether males adjust incubation effort to the mating context and to examine how females respond to their partner’s effort. Birds without previous breeding experience were paired randomly and bred with the same partner twice. In the first breeding attempt, half the males received EPC opportunities with ‘extra-pair females’ during incubation, while the other half did not. Males that received EPC opportunities in the first breeding attempt did not in the second breeding attempt and vice versa. We recorded incubation effort on days when EPC opportunities were not presented. In their first breeding attempt, males with EPC opportunities incubated less than those without. Females compensated fully for the deficit in male care so that a pair’s combined incubation effort was unchanged. In the second attempt, when a male’s opportunity for EPCs was switched, individuals showed the same level of incubation effort that they had previously, irrespective of the current availability of extra-pair females. This suggests that division of effort was negotiated in the first breeding attempt and maintained without significant adjustments in the second attempt. The effects of male EPC opportunity in the first breeding attempt on subsequent incubation effort suggests that individual parental decisions can be shaped by previous experience and this may partly explain conflicting results in studies where individuals’ histories were not known.

Ginkgo biloba extract enhances noncontact erection in rats: the role of dopamine in the paraventricular nucleus and the mesolimbic system

Penile erection is essential for successful copulation in males. Dopaminergic projections from the paraventricular nucleus (PVN) to the ventral tegmental area (VTA) and from the VTA to the nucleus accumbens (NAc) are thought to exert a facilitatory effect on penile erection. Our previous study showed that treatment with an extract of Ginkgo biloba leaves (EGb 761) enhances noncontact erection (NCE) in male rats. However, the relationship between NCE and dopaminergic activity in the PVN, VTA, and NAc remains unknown. The present study examined the relationship between NCE and central dopaminergic activity following EGb 761 treatment. We report here that, in comparison with the controls, there was a significant increase in the number of NCEs in rats after treatment with 50 mg/kg of EGb 761 for 14 days. EGb 761-treated rats also showed more NCEs than the same group before EGb 761 treatment. A significant increase in the expression of catecholaminergic neurons in the PVN and the VTA was seen by means of tyrosine hydroxylase immunohistochemistry, and tissue levels of dopamine and 3,4-dihydroxyphenylacetic acid in the NAc were also markedly increased in the EGb 761-treated animals. However, the norepinephrine tissue levels in the PVN and the NAc in the EGb 761-treated group were not significantly different from those in the controls. Together, these results suggest that administration of EGb 761 increases dopaminergic activity in the PVN and the mesolimbic system to facilitate NCE in male rats.

Male wing fanning by the aphid parasitoidAphidius ervi(Hymenoptera: Braconidae) produces a courtship song

We, herein, report evidence that wing fanning by the aphid parasitoid Aphidius ervi Haliday (Hymenoptera: Braconidae) constitutes a courtship song. Complete removal of the forewings or only the distal half of them reduced male copulation success in comparison to intact males. Males that achieved copulation within the observation period produced wing fanning at a higher rate than males that did not copulate. Playback of wing fanning sound altered the behaviour of virgin females, increasing the time they devoted to grooming, as compared with subjects that were exposed to silence or white noise. Our findings contribute to the understanding of the acoustic dimension of the sensory modalities employed by this aphid parasitoid in sexual signalling.

Mate choice opportunity leads to shorter offspring development time in a desert insect

We describe indirect genetic benefits of mate choice in two allopatric populations of cactophilic Drosophila mojavensis. By manipulating mate choice opportunity, we show that greater mate choice among sexually mature adults leads to shorter offspring egg-to-adult development times; the extent of this reduction was influenced by population origin and by host plant environment. We performed multiple-choice mating trials with individually marked flies to investigate whether differential male mating success was a consequence of female choice, male interaction, or both. We demonstrate that male copulation frequency was not random and instead, was determined by female choice. Virgin females in these trials were no less discriminating than females that had been previously exposed to males. These results suggest that there are indirect benefits of female mate choice that are population and environment specific, consistent with the hypothesis of ecologically influenced 'good genes' sexual selection.

A Cholinergic-Regulated Circuit Coordinates the Maintenance and Bi-Stable States of a Sensory-Motor Behavior during Caenorhabditis elegans Male Copulation

Penetration of a male copulatory organ into a suitable mate is a conserved and necessary behavioral step for most terrestrial matings; however, the detailed molecular and cellular mechanisms for this distinct social interaction have not been elucidated in any animal. During mating, the Caenorhabditis elegans male cloaca is maintained over the hermaphrodite's vulva as he attempts to insert his copulatory spicules. Rhythmic spicule thrusts cease when insertion is sensed. Circuit components consisting of sensory/motor neurons and sex muscles for these steps have been previously identified, but it was unclear how their outputs are integrated to generate a coordinated behavior pattern. Here, we show that cholinergic signaling between the cloacal sensory/motor neurons and the posterior sex muscles sustains genital contact between the sexes. Simultaneously, via gap junctions, signaling from these muscles is transmitted to the spicule muscles, thus coupling repeated spicule thrusts with vulval contact. To transit from rhythmic to sustained muscle contraction during penetration, the SPC sensory-motor neurons integrate the signal of spicule's position in the vulva with inputs from the hook and cloacal sensilla. The UNC-103 K+ channel maintains a high excitability threshold in the circuit, so that sustained spicule muscle contraction is not stimulated by fewer inputs. We demonstrate that coordination of sensory inputs and motor outputs used to initiate, maintain, self-monitor, and complete an innate behavior is accomplished via the coupling of a few circuit components.