Ejaculate Allocation and Sperm Characteristics Differ among Alternative Male Types in a Species of Fish with Cooperation and Competition among Unrelated Males

Sexual selection arising from sperm competition has driven the evolution of immense variation in ejaculate allocation and sperm characteristics not only among species, but also among males within a species. One question that has received little attention is how cooperation among males affects these patterns. Here we ask how male alternative reproductive types differ in testes size, ejaculate production, and sperm morphology in the ocellated wrasse, a marine fish in which unrelated males cooperate and compete during reproduction. Nesting males build nests, court females and provide care. Sneaker males only “sneak” spawn, while satellite males sneak, but also help by chasing away sneakers. We found that satellite males have larger absolute testes than either sneakers or nesting males, despite their cooperative role. Nesting males invested relatively less in testes than either sneakers or satellites. Though sneakers produced smaller ejaculates than either satellite or nesting males, we found no difference among male types in either sperm cell concentration or sperm number, implying sneakers may produce less seminal fluid. Sperm tail length did not differ significantly among male types, but sneaker sperm cells had significantly larger heads than either satellite or nesting male sperm, consistent with past research showing sneakers produce slower sperm. Our results highlight that social interactions among males can influence sperm and ejaculate production.

In search of an honest butterfly: sexually selected wing coloration and reproductive traits from wild populations of the cabbage white butterfly

Sexual selection is an important and well-studied topic and is central to many theories on mate selection and individual behavior. Relatively little is known about the impacts that human-induced rapid environmental change are having on secondary sexually selected characteristics. In particular, we lack a clear understanding of the effects of the introduction of once-limiting nutrients to wild populations. Honest signals function as an indicator of mate quality when there are differences in nutrient acquisition and allocation to secondary traits. We used the cabbage white butterfly, Pieris rapae (L.), to investigate differences in color and testes size as an index of reproductive potential. We collected individuals from four sites in California and Nevada to capture variation among areas with high and low cropland and potential nitrogen availability. Differences in mean testes size among sites raise the possibility that individuals from sites surrounded by high agricultural areas have smaller testes than individuals from less agriculturally-developed areas. Coloration variables and testes size were positively associated, consistent with the hypothesis that nitrogen-based coloration in the cabbage white is an honest mating signal. However, variation among sites in that relationship suggests complexities that need further exploration, including the possibility that the signal is not of equal value in all populations. Thus these results advance our understanding of complex relationships among human-induced environmental change and sexual selection in the wild.

Control of testes mass by androgen receptor paralogs in a cichlid

Steroid hormones play numerous important and diverse roles in the differentiation and development of vertebrate primary and secondary reproductive characteristics. However, the exact role of androgen receptors (ARs)—which bind circulating androgens—in this regulatory pathway is unclear. Teleost fishes further complicate this question by having two paralogs of AR (ARα and ARβ) resulting from a duplication of their ancestral genome. We investigated the functional role of these two ARs on testes growth and development by experimentally eliminating receptor function of one or both paralogs using CRISPR/Cas9 genome edited Astatotilapia burtoni, an African cichlid fish. Fish with two or more functional receptor alleles were more likely to be male compared to fish with one or fewer, suggesting that the two paralogs of the receptor may be redundant in regulating early sex determination. In contrast, we found that adult testes size was significantly affected by distinct combinations of mutant and wild-type AR alleles. We present a working model whereby ARβ facilitates testes growth and ARα causes testes regression. This mechanism may contribute to the robust social and physiological plasticity displayed by A. burtoni and other social teleost fish.

Patterns of Gonadic Development and Operational Sex Ratio Promote Promiscuity of Euxesta bilimeki (Diptera:Ulidiidae)

Promiscuous mating systems are widely distributed among animals and can be promoted by operational sex ratios (number of receptive adults; OSR). In populations where OSR is not biased towards any sex, the possibility that males and females mate with several individuals increases. For both sexes to synchronize in time and space for reproduction, adults should possess or simultaneously acquire nutrients required to reach sexual maturity. Among synovigenic (without a full complement of eggs at eclosion) species, nutrient acquisition, protein in particular, may influence the OSR. In the Agave fly Euxesta bilimeki (Hendel) (Diptera: Ulidiidae), both sexes engage in multiple mating and females frequently expel all or part of the ejaculate. Here, we assessed the effect of protein intake on gonadic development, and estimated OSR from field-collected individuals. Body protein content was compared between wild and laboratory individuals with access to different diets, and mating frequency and individual mating rate were analyzed for cohorts at a 1:1 sex ratio. Both sexes required protein ingestion for gonadic development, but there were no differences in protein content between field-collected males and males fed protein and sugar in the laboratory, despite the fact that males assigned 9.3% of their corporal weight to testicles. Euxesta bilimeki is a promiscuous species where both males and females mate multiply with one or several individuals in short periods of time; thus, large testes size may be linked to the need of voluminous ejaculate production, and might be further exacerbated by female ejaculate expulsion.

Seasonal variation in gonad physiology indicates juvenile breeding in the Saker Falcon (Falco cherrug)

We describe seasonal changes in the physiology of reproductive organs of Saker Falcons electrocuted on an electricity power line in Mongolia. Macroscopic examination of the gonads revealed asymmetry in testes size, with bias to the left body side. This asymmetry declined with seasonal increase in testes size during the main egg-laying period of the breeding season. In females, 91% possessed both left and right ovaries ( N = 161); there was no visible oviduct associated with the right ovary and it was smaller than the functional left ovary. Both ovaries showed seasonal development in size, becoming larger during the peak egg-laying period. Hierarchical preovulatory follicular development was recorded in two females, with ca. 5 mm difference in the diameter of sequential follicles. Both sexes developed brood patches during the main incubation period, with adults more likely to exhibit brood patches than juveniles. Among juveniles, at least 82% of females and 91% of males were non-breeders without brood patches. The high electrocution rate at our studied power line provided a rare opportunity to examine the non-breeding component of the Saker population. Juveniles predominated in the non-breeding population during the main egg-laying period (89%, N = 65), with the proportion of adults electrocuted being significantly lower among females. Only a small proportion of juvenile females exhibited gonadal evidence of breeding, consistent with the low observed frequency of juvenile breeders at nests. The demographic composition of the non-breeding population is consistent with female mortality rates exceeding that of males, and potentially indicates incipient population decline.

How sperm competition shapes the evolution of testes and sperm: a meta-analysis

Females of many species mate with multiple males, thereby inciting competition among ejaculates from rival males for fertilization. In response to increasing sperm competition, males are predicted to enhance their investment in sperm production. This prediction is so widespread that testes size (correcting for body size) is commonly used as a proxy of sperm competition, even in the absence of any other information about a species' reproductive behaviour. By contrast, a debate about whether sperm competition selects for smaller or larger sperm has persisted for nearly three decades, with empirical studies demonstrating every possible response. Here, we synthesize nearly 40 years of sperm competition research in a meta-analytical framework to determine how the evolution of sperm number (i.e. testes size) and sperm size (i.e. sperm head, midpiece, flagellum and total length) is influenced by varying levels of sperm competition across species. Our findings support the long-held assumption that higher levels of sperm competition are associated with relatively larger testes. We also find clear evidence that sperm competition is associated with increases in all components of sperm length. We discuss these results in the context of different theoretical predictions and general patterns in the breeding biology and selective environment of sperm. This article is part of the theme issue ‘Fifty years of sperm competition’.

Extensive geographical variation in testes size and ejaculate traits in a terrestrial-breeding frog

Ejaculate traits vary extensively among individuals and species, but little is known about their variation among populations of the same species. Here, we investigated patterns of intraspecific variation in male reproductive investment in the terrestrial-breeding frog Pseudophryne guentheri . Like most anurans, breeding activity in P. guentheri is cued by precipitation, and therefore the timing and duration of breeding seasons differ among geographically separated populations, potentially leading to differences in the level of sperm competition. We, therefore, anticipated local adaptation in sperm traits that reflect these phenological differences among populations. Our analysis of six natural populations across a rainfall gradient revealed significant divergence in testes and ejaculate traits that correspond with annual rainfall and rainfall seasonality; males from the northern and drier edge of the species range had significantly smaller testes containing fewer, smaller and less motile sperm compared with those from mesic central populations. These findings may reflect spatial variation in the strength of postcopulatory sexual selection, likely driven by local patterns of precipitation.