Pupal Cues Increase Sperm Production but Not Testis Size in an Insect

Theoretic and empirical studies show that social surroundings experienced by male insects during their larval or adult stage can influence their testicular investment in diverse ways. Although insect pupae do not feed and crawl, they can communicate using sex-specific and/or non-sex specific cues. Yet, it is unknown, in any insect, whether and how male pupae can fine-tune their resource allocation to sperm production and testis size in response to socio-sexual environments. We investigated this question using a moth, Ephestia kuehniella, which produces fertile eupyrene sperm and unfertile apyrene sperm. We held male pupae individually or in groups with different sex ratios, and dissected adults upon eclosion, measured their testis size, and counted both types of sperm. We demonstrated that after exposure to conspecific pupal cues regardless of sex, male pupae increased production of eupyrenes and apyrenes at the same rate but kept testis size unchanged. We suggest that testis size is fixed after pupation because most morphological traits are formed during the larval stage, allowing little room for pupae to adjust testis size. Like adults, male pupae with fully grown testes have sufficient resources to produce more sperm of both types according to the perceived increase in sperm competition risk.

Pupal Cues Increase Sperm Production but Not Testis Size in an Insect

Theoretic and empirical studies show that social surroundings experienced by male insects during their larval or adult stage can influence their testicular investment in diverse ways. Although insect pupae do not feed and crawl, they can communicate using sex-specific and/or non-sex specific cues. Yet, it is unknown, in any insect, whether and how male pupae can fine-tune their resource allocation to sperm production and testis size in response to socio-sexual environment. We investigated this question using a moth, Ephestia kuehniella, which produces fertile eupyrene sperm and unfertile apyrene sperm. We held male pupae individually or in groups with different sex ratios, and dissected adults upon eclosion, measured their testis size, and counted both types of sperm. We demonstrated that after exposure to conspecific pupal cues regardless of sex, male pupae increased production of eupyrenes and apyrenes at the same rate but kept testis size unchanged. We suggest that testis size is fixed after pupation because most morphological traits are formed during the larval stage, allowing little room for pupae to adjust testis size. Like adults, male pupae with fully grown testes have sufficient resources to produce more sperm of both types according to the perceived increase of sperm competition risk.

Males adjust their manipulation of female remating in response to sperm competition risk

To reduce the potential for sperm competition, male insects are thought to inhibit the post-mating reproductive behaviour of females through receptivity-inhibiting compounds transferred in the ejaculate. Selection is expected to favour phenotypic plasticity in male post-copulatory expenditure, with males investing strategically in response to their perceived risk of sperm competition. However, the impact that socially cued strategic allocation might have on female post-mating behaviour has rarely been assessed. Here, we varied male perception of sperm competition risk, both prior to and during mating, to determine if a male's competitive environment impacts the extent to which he manipulates female remating behaviour. We found that female Australian field crickets ( Teleogryllus oceanicus ) mated to males that were reared under sperm competition risk emerged from a shelter in search of male song sooner than did females mated to males reared without risk, but only when mating occurred in a risk-free environment. We also found that females reared in a silent environment where potential mates were scarce emerged from the shelter sooner than females exposed to male calls during development. Collectively, our findings suggest complex interacting effects of male and female sociosexual environments on female post-mating sexual receptivity.

Spatially clustered resources increase aggregation and mating duration in Drosophila melanogaster

ABSTRACTIn environments where females mate multiply, males should adjust their behaviour and physiology in response to the prevailing perceived level of sperm competition. This expectation is well supported by both laboratory and field studies, but we don’t yet know what mechanisms facilitate these plastic responses in natural populations. One way in which males appear to assess sperm competition risk is through encounter rates with conspecific males. Such encounter rates may be driven by the spatial distribution of resources required by male. However, explicit links between resource distribution, male encounter rate, and shifts in behaviour related to sperm competition have not been demonstrated. Here we show that a small increase in the distance of patches of food resources in the laboratory: (a) approximately halves the mean distances between pairs of males; and (b) is associated with an increase in subsequent copulation duration – previously shown to be a reliable indicator of male perception of sperm competition risk – by more than two minutes. Aggregation of resources, operating via increased encounter rate, is a mechanism that can stimulate plastic male sperm competition responses. Because spatial distribution of resources, including those exploited by Drosophila, is variable in nature, this may explain one way in which sperm competition-related plasticity is influenced in wild-living males.

Effects of condition and sperm competition risk on sperm allocation and storage in neriid flies

Ejaculate traits can be sexually selected and often exhibit heightened condition-dependence. However, the influence of sperm competition risk in tandem with condition-dependent ejaculate allocation strategies is relatively unstudied. Because ejaculates are costly to produce, high-condition males may be expected to invest more in ejaculates when sperm competition risk is greater. We examined the condition-dependence of ejaculate size by manipulating nutrient concentration in the juvenile (larval) diet of the neriid fly Telostylinus angusticollis. Using a fully factorial design we also examined the effects of perceived sperm competition risk (manipulated by allowing males to mate first or second) on the quantity of ejaculate transferred and stored in the three spermathecae of the female reproductive tract. To differentiate male ejaculates, we fed males nontoxic rhodamine fluorophores (which bind to proteins in the body) prior to mating, labeling their sperm red or green. We found that high-condition males initiated mating more quickly and, when mating second, transferred more ejaculate to both of the female’s posterior spermathecae. This suggests that males allocate ejaculates strategically, with high-condition males elevating their ejaculate investment only when facing sperm competition. More broadly, our findings suggest that ejaculate allocation strategies can incorporate variation in both condition and perceived risk of sperm competition.

Male mate choice for large gravid spots in a livebearing fish

Male mate choice occurs in a wide range of species, and males can increase their reproductive success by distinguishing between females based on their fecundity (e.g., large body size) or their expected sperm competition risk (e.g., virgins). However, patterns of male mate choice could be mitigated by variation in female physiological receptivity, as males can benefit by directing their mating efforts toward females that are at a point in their reproductive cycle when fertilization probability is highest. Here, we perform three experiments to assess whether male mate choice is influenced by cues of female physiological receptivity, fecundity, or sperm competition risk in the pygmy halfbeak (Dermogenys collettei), a small livebearing fish. Female halfbeaks possess a “gravid spot”—an orange abdominal marking that is caused by pigmentation of the females’ skin and variation in embryo development and pigmentation during pregnancy. We show that gravid spot size increases toward parturition and is largest right before giving birth, independent of abdominal width or body size. Males consistently chose females with large gravid spots over females with small gravid spots. In contrast, males did not prefer larger females over smaller females or virgin females over mated females. As female halfbeaks store sperm prior to fertilizations, we suggest that males use the size of the gravid spot as a cue to direct their mating efforts to those females where the chance of fertilization is highest.

Strategic adjustment of copulatory plug size in a nematode

Copulatory plugs (CP) are substances produced during copulation that block the genital openings of the female. In several species of Nematoda, males produce CP that are thought to impede female remating and thus sperm competition. The relatively large size of the CP in several nematodes, and its evolutionary loss in self-fertilizing populations of Caenorhabditis elegans, suggests that CP are costly to produce. If CP production is costly, the application of basic concepts of strategic ejaculation theory suggests a modulated allocation of CP in response to sperm competition risk. This hypothesis led us to predict that males perceiving a higher risk of sperm competition will produce larger CP. We tested these ideas with the entomopathogenic, gonochoristic nematode Rhabditis regina. Our first experiment provides evidence suggesting that production of CP is costly, because the size of CP is negatively affected by stressful conditions (high population density, small male adult size, and suboptimal food type). The results of our second experiment support the prediction that males adjust the size of CP to sperm competition risk: the average size of CP increased as the number of males competing for one female increased. Overall, our study supports the idea that in R. regina the production of CP is costly for males and that the size of the CP produced is influenced by sperm competition risk.