Sperm competition risk drives plasticity in seminal fluid composition.

BACKGROUND: Ejaculates contain a diverse mixture of sperm and seminal fluid proteins, the combination of which is crucial to male reproductive success under competitive conditions. Males should therefore tailor the production of different ejaculate components according to their social environment, with particular sensitivity to cues of sperm competition risk (i.e. how likely it is that females will mate promiscuously). Here we test this hypothesis using an established vertebrate model system, the house mouse (Mus musculus domesticus), combining experimental data with a quantitative proteomics analysis of seminal fluid composition. Our study tests for the first time how both sperm and seminal fluid components of the ejaculate are tailored to the social environment. RESULTS: Our quantitative proteomics analysis reveals that the relative production of different proteins found in seminal fluid--i.e. seminal fluid proteome composition--differs significantly according to cues of sperm competition risk. Using a conservative analytical approach to identify differential expression of individual seminal fluid components, at least seven of 31 secreted seminal fluid proteins examined showed consistent differences in relative abundance under high versus low sperm competition conditions. Notably three important proteins with potential roles in sperm competition--SVS 6, SVS 5 and CEACAM 10--were more abundant in the high competition treatment groups. Total investment in both sperm and seminal fluid production also increased with cues of heightened sperm competition risk in the social environment. By contrast, relative investment in different ejaculate components was unaffected by cues of mating opportunities. CONCLUSIONS: Our study reveals significant plasticity in different ejaculate components, with the production of both sperm and non-sperm fractions of the ejaculate strongly influenced by the social environment. Sperm competition risk is thus shown to be a key factor in male ejaculate production decisions, including driving plasticity in seminal fluid composition.

Sexual conflict and sperm competition.

Traits that increase a male's fertilization success during sperm competition can be harmful to females and therefore represent a source of sexual conflict. In this review, we consider the variety of male adaptations to sperm competition (MASC) that may give rise to sexual conflict-including mate guarding, prolonged copulations, the transfer of large numbers of sperm, and the manipulation of females through nonsperm components of the ejaculate. We then reflect on the fitness economics influencing the escalation of these sexual conflicts, considering the likelihood of females evolving traits to offset the negative effects of MASC when compared with the strong selection on males that lead to MASC. We conclude by discussing the potential evolutionary outcomes of sexual conflict arising from MASC, including the opportunities for females to mitigate conflict costs and the prospects for conflict resolution.

Sexual conflict and interacting phenotypes: a quantitative genetic analysis of fecundity and copula duration in Drosophila melanogaster.

Many reproductive traits that have evolved under sexual conflict may be influenced by both sexes. Investigation of the genetic architecture of such traits can yield important insight into their evolution, but this entails that the heritable component of variation is estimated for males and females-as an interacting phenotype. We address the lack of research in this area through an investigation of egg production and copula duration in the fruit fly, Drosophila melanogaster. Despite egg production rate being determined by both sexes, which may cause sexual conflict, an assessment of this trait as an interacting phenotype is lacking. It is currently unclear whether copula duration is determined by males and/or females. We found significant female, but not male, genetic variance for egg production rate that may indicate reduced potential for ongoing sexually antagonistic coevolution. In contrast, copula duration was determined by significant genetic variance in both sexes. We also identified genetic variation in egg retention among virgin females. Although previously identified in wild populations, it is unclear why this should be present in a laboratory stock. This study provides a novel insight into the shared genetic architecture of reproductive traits that are the subject of sexual conflict.

Genome-Wide Responses of Female Fruit Flies Subjected to Divergent Mating Regimes.

Elevated rates of mating and reproduction cause decreased female survival and lifetime reproductive success across a wide range of taxa from flies to humans. These costs are fundamentally important to the evolution of life histories. Here we investigate the potential mechanistic basis of this classic life history component. We conducted 4 independent replicated experiments in which female Drosophila melanogaster were subjected to 'high' and 'low' mating regimes, resulting in highly significant differences in lifespan. We sampled females for transcriptomic analysis at day 10 of life, before the visible onset of ageing, and used Tiling expression arrays to detect differential gene expression in two body parts (abdomen versus head+thorax). The divergent mating regimes were associated with significant differential expression in a network of genes showing evidence for interactions with ecdysone receptor. Preliminary experimental manipulation of two genes in that network with roles in post-transcriptional modification (CG11486, eyegone) tended to enhance sensitivity to mating costs. However, the subtle nature of those effects suggests substantial functional redundancy or parallelism in this gene network, which could buffer females against excessive responses. There was also evidence for differential expression in genes involved in germline maintenance, cell proliferation and in gustation / odorant reception. Interestingly, we detected differential expression in three specific genes (EcR, keap1, lbk1) and one class of genes (gustation / odorant receptors) with previously reported roles in determining lifespan. Our results suggest that high and low mating regimes that lead to divergence in lifespan are associated with changes in the expression of genes such as reproductive hormones, that influence resource allocation to the germ line, and that may modify post-translational gene expression. This predicts that the correct signalling of nutrient levels to the reproductive system is important for maintaining organismal integrity.

Variation in male mate choice in Drosophila melanogaster.

Male mate choice has been reported in the fruit fly, Drosophila melanogaster, even though males of this species were previously thought to maximise their fitness by mating with all available females. To understand the evolution of male mate choice it is important to understand variation in male mating preferences. Two studies, using different stock populations and different methods, have reported contrasting patterns of variation in male mate choice in D. melanogaster. Two possible explanations are that there are evolved differences in each stock population or that the methods used to measure choice could have biased the results. We investigated these hypotheses here by repeating the methods used in one study in which variable male mate choice was found, using the stock population from the other study in which choice was not variable. The results showed a significant resource-independent male preference for less fecund, smaller females, which contrasts with previous observations of male mate choice. This indicates that different selection pressures between populations have resulted in evolved differences in the expression of male mate choice. It also reveals phenotypic plasticity in male mate choice in response to cues encountered in each choice environment. The results highlight the importance of variation in male mate choice, and of identifying mechanisms in order to understand the evolution of mate choice under varying ecological conditions.

Male-specific genotype by environment interactions influence viability selection acting on a sexually selected inversion system in the seaweed fly, Coelopa frigida.

In the seaweed fly, Coelopa frigida, a large chromosomal inversion system is affected by sexual selection and viability selection. However, our understanding of the interaction between these two selective forces is currently limited as research has focused upon a limited range of environments. We allowed C. frigida larvae to develop in two different algae, Fucus and Laminaria, and then measured viability and body size for each inversion genotype. Significant male-specific genotype-by-environment interactions influenced viability and body size. For males developing in Laminaria, the direction of viability selection acts similarly on the inversion system as the direction of sexual selection. In contrast, for males developing in Fucus, viability selection opposes sexual selection. These results demonstrate that through considering viability selection in different environments, the costs and benefits associated with sexual selection can be found to vary.

Sex-specific effects of developmental environment on reproductive trait expression in Drosophila melanogaster.

Variation in the expression of reproductive traits provides the raw material upon which sexual selection can act. It is therefore important to understand how key factors such as environmental variation influence the expression of reproductive traits, as these will have a fundamental effect on the evolution of mating systems. It is also important to consider the effects of environmental variation upon reproductive traits in both sexes and to make comparisons with the environment to which the organism is adapted. In this study, we addressed these issues in a systematic study of the effect of a key environmental factor, variation in larval density, on reproductive trait expression in male and female Drosophila melanogaster. To do this, we compared reproductive trait expression when flies were reared under controlled conditions at eight different larval densities that covered a 20-fold range. Then, to place these results in a relevant context, we compared the results to those from flies sourced directly from stock cages. Many reproductive traits were surprisingly insensitive to variation in larval density. A notable exception was nonlinear variation in female fecundity. In contrast, we found much bigger differences in comparisons with flies from stock cages-including differences in body size, latency to mate, copulation duration, fecundity, and male share of paternity in a competitive environment. For a number of traits, even densities of 1000 larvae per vial (125 larvae per mL of food) did not phenocopy stock cage individuals. This study reveals novel patterns of sex-specific sensitivity to environmental variation that will influence the strength of sexual selection. It also illustrates the importance of comparisons with the environment to which individuals are adapted.

Measuring the fitness benefits of male mate choice in Drosophila melanogaster.

It is increasingly realized that the potential for male mate choice is widespread across many taxa. However, measurements of the relative magnitude of the fitness benefits that such choice can confer are lacking. Here, we directly measured, in a comprehensive set of tests that manipulated key variables, the fitness benefits of male mate choice in Drosophila melanogaster by measuring egg production in females that were chosen or rejected by males. The results provided significant evidence for male mate choice. In absolute terms, the observed degree of choice increased male fitness by an average of only 1.59 eggs. However, using a novel technique we show that this benefit of choice represented 14.5% of the maximum potential fitness benefit of choice. The magnitude of mate choice was not significantly altered by variation in (1) mate compatibility, (2) phenotypic plasticity in male mate choice, or (3) whether choosing males were preferred or nonpreferred by females. Overall, we show that male mate choice represents a subtle but significant opportunity for sexual selection, and we offer a novel and widely applicable method for quantifying mate choice.

Interactions between genotype and sexual conflict environment influence transgenerational fitness in Drosophila melanogaster.

Theory predicts that sexual conflict can fuel evolutionary change and generate substantial reproductive costs. This was tested here by measuring the fitness of focal individuals across multiple generations using an experimental framework. We manipulated sexual conflict through high versus low exposure of females to males across a four-generation pedigree of Drosophila melanogaster, and assessed fitness in 1062 females and 639 males. We used the animal model to estimate (1) genotype by sexual conflict environment interactions for female fitness and (2) indirect benefits gained through sons and daughters. Some female genotypes achieved higher fitness under low, in comparison to high, conflict and vice versa. We found a consistent 10% reduction in female fitness under high conflict, regardless of maternal history. Following high exposure, females produced sons with increased, but grandsons with decreased, fitness. This opposing effect suggests no consistent fitness gains through sons for females that mated multiply. We saw no indirect benefits through daughters. Our pedigree was based exclusively on maternal links; however, maternal effects are unlikely to contribute significantly unless expressed across multiple generations. In sum, we quantified a significant sexual conflict load and a female genotype by sexual conflict interaction that could slow the erosion of genetic variation.

The evolution and significance of male mate choice.

The distinct reproductive roles of males and females, which for many years were characterised in terms of competitive males and choosy females, have remained a central focus of sexual selection since Darwin's time. Increasing evidence now shows that males can be choosy too, even in apparently unexpected situations, such as under polygyny or in the absence of male parental care. Here, we provide a synthesis of the theory on male mate choice and examine the factors that promote or constrain its evolution. We also discuss the evolutionary significance of male mate choice and the contrasts in male versus female mate choice. We conclude that mate choice by males is potentially widespread and has a distinct role in how mating systems evolve.

Quantifying the life-history response to increased male exposure in female Drosophila melanogaster.

Precise estimates of costs and benefits, the fitness economics, of mating are of key importance in understanding how selection shapes the coevolution of male and female mating traits. However, fitness is difficult to define and quantify. Here, we used a novel application of an established analytical technique to calculate individual- and population-based estimates of fitness-including those sensitive to the timing of reproduction-to measure the effects on females of increased exposure to males. Drosophila melanogaster females were exposed to high and low frequencies of contact with males, and life-history traits for each individual female were recorded. We then compared different fitness estimates to determine which of them best described the changes in life histories. We predicted that rate-sensitive estimates would be more accurate, as mating influences the rate of offspring production in this species. The results supported this prediction. Increased exposure to males led to significantly decreased fitness within declining but not stable or increasing populations. There was a net benefit of increased male exposure in expanding populations, despite a significant decrease in lifespan. The study shows how a more accurate description of fitness, and new insights can be achieved by considering individual life-history strategies within the context of population growth.

Adaptations to sexual selection and sexual conflict: insights from experimental evolution and artificial selection.

Artificial selection and experimental evolution document natural selection under controlled conditions. Collectively, these techniques are continuing to provide fresh and important insights into the genetic basis of evolutionary change, and are now being employed to investigate mating behaviour. Here, we focus on how selection techniques can reveal the genetic basis of post-mating adaptations to sexual selection and sexual conflict. Alteration of the operational sex ratio of adult Drosophila over just a few tens of generations can lead to altered ejaculate allocation patterns and the evolution of resistance in females to the costly effects of elevated mating rates. We provide new data to show how male responses to the presence of rivals can evolve. For several traits, the way in which males responded to rivals was opposite in lines selected for male-biased, as opposed to female-biased, adult sex ratio. This shows that the manipulation of the relative intensity of intra- and inter-sexual selection can lead to replicable and repeatable effects on mating systems, and reveals the potential for significant contemporary evolutionary change. Such studies, with important safeguards, have potential utility for understanding sexual selection and sexual conflict across many taxa. We discuss how artificial selection studies combined with genomics will continue to deepen our knowledge of the evolutionary principles first laid down by Darwin 150 years ago.

Sex-biased phoretic mite load on two seaweed flies: Coelopa frigida and Coelopa pilipes.

Two hypotheses explain male-biased parasitism. Physiological costs of male sexually selected characteristics can reduce immunocompetence. Alternatively, ecological differences could generate male-biased parasitism. One method of comparing the importance of the two theories is to investigate patterns of phoresy, which are only likely to be generated by ecological rather than immunological differences between the sexes. Here we studied the pattern of phoresy of the mite, Thinoseius fucicola, on two species of seaweed fly hosts, Coelopa frigida and Coelopa pilipes. We found a highly male-biased pattern of phoresy of T. fucicola on both species. These are the first reported instances of sex-biased phoresy in a solely phoretic parasite. We also show the first two cases of size-biased phoresy. We suggest that ecological factors, particularly, male mate searching, generated male biased patterns of phoresy. We highlight the potential importance of studies of phoresy in determining the relative roles of the immunocompetence and ecological theories in generating male-biased parasitism. We suggest that more studies of patterns of phoresy are carried out to allow detailed comparisons with patterns of parasitism.