Mating for male-derived prostaglandin: a functional explanation for the increased fecundity of mated female crickets?

Direct benefits are considered to be the driving force of high female mating rates, yet species in which females do not receive material resources from males still experience increased fitness from mating frequently. One hypothesis suggests that substances within the ejaculate may boost survival or offspring production. If these materials are limiting to females, they will require continual renewal via mating and could provide a functional understanding of how high mating rates lead to increased female fitness. Using the Texas field cricket, Gryllus texensis, we investigated the sexual transfer of prostaglandin E2, an important mediator of invertebrate reproduction. We determined that like other gryllid species, males include significant quantities of prostaglandin E2 (PGE2) and its precursor molecule, arachidonic acid (AA), within the spermatophore. These components are passed to females during copulation and then stored within the spermatheca. We then tested the novel hypothesis that PGE2 is ephemerally available after mating and that females must frequently mate to maintain access to this limiting compound. We found that PGE2 within the spermatheca is indeed depleted through time, with only a small amount remaining 1 week after mating, but that its presence can be maintained at high quantities and for prolonged periods of time by remating. Our results support the hypothesis that high female mating rates increase the amount and availability of PGE2 throughout the breeding season, which could explain the positive relationship between female mating rate and fecundity.

Phenotypically plastic responses to environmental variation are more complex than life history theory predicts.

For insects that exhibit wing polyphenic development, abiotic and biotic signals dictate the adult wing morphology of the insect in an adaptive manner such that in stressful environments the formation of a flight-capable morph is favored and in low-stress environments, a flightless morph is favored. While there is a relatively large amount known about the environmental cues that dictate morph formation in wing polyphenic hemipterans like planthoppers and aphids, whether those cues dictate the same morphs in non-hemipteran (i.e., cricket) wing polyphenic species has not been explicitly investigated. To experimentally test the generality of environmental cue determination of wing polyphenism across taxa with diverse life histories, in this study, we tested the importance of food quantity, parasitic infection, and tactile cues on wing morph determination in the wing polyphenic sand field cricket, Gryllus firmus. Our results also show that certain stress cues, such as severe diet quantity limitation and parasitic infection, actually led to an increase in the production of flightless morph. Based on these findings, our results suggest that physiological and genetic constraints are important to an organism's ability to respond to environmental variation in an adaptive manner beyond simple life history trade-offs.

The developmental high wire: Balancing resource investment in immunity and reproduction.

The strategic allocation of resources into immunity poses a unique challenge for individuals, where infection at different stages of development may result in unique trade-offs with concurrent physiological processes or future fitness-enhancing traits. Here, we experimentally induced an immune challenge in female Gryllus firmus crickets to test whether illness at discrete life stages differentially impacts fitness. We injected heat-killed Serratia marcescens bacteria into antepenultimate juveniles, penultimate juveniles, sexually immature adults, and sexually mature adults, and then measured body growth, instar duration, mating rate, viability of stored sperm, egg production, oviposition rate, and egg viability. Immune activation significantly impacted reproductive traits, where females that were immune challenged as adults had decreased mating success and decreased egg viability compared to healthy individuals or females that were immune challenged as juveniles. Although there was no effect of an immune challenge on the other traits measured, the stress of handling resulted in reduced mass gain and smaller adult body size in females from the juvenile treatments, and females in the adult treatments suffered from reduced viability of sperm stored within their spermatheca. In summary, we found that an immune challenge does have negative impacts on reproduction, but also that even minor acute stressors can have significant impacts on fitness-enhancing traits. These findings highlight that the factors affecting fitness can be complex and at times unpredictable, and that the consequences of illness are specific to when during an individual's life an immune challenge is induced.

Fluctuating Asymmetry in the Polymorphic Sand Cricket (Gryllus firmus): Are More Functionally Important Structures Always More Symmetric?

Fluctuating asymmetry (FA) may serve as a reliable indicator of the functional importance of structures within an organism. Primary locomotor structures often display lower levels of FA than other paired structures, highlighting that selection can maintain symmetry in fitness-enhancing traits. Polyphenic species represent an attractive model for studying the fine-scale relationship between trait form and function, because multiple morphs exhibit unique life history adaptations that rely on different traits to maximize fitness. Here, we investigated whether individuals of the wing polyphenic sand field cricket (Gryllus firmus) maintain higher levels of symmetry in the bilateral structures most vital for maximizing fitness based on their specific life history strategy. We quantified FA and directional asymmetry (DA) across a suite of key morphological structures indicative of investment in somatic growth, reproduction, and flight capability for males and females across the flight-capable longwing (LW) and flight-incapable shortwing (SW) morphs. Although we did not find significant differences in FA across traits, hindwings lacked DA that was found in all other structures. We predicted that functionally important traits should maintain a higher level of symmetry; however, locomotor compensation strategies may reduce the selective pressures on symmetry or developmental constraints may limit the optimization between trait form and function.

The Fat-Dachsous signaling pathway regulates growth of horns in Trypoxylus dichotomus, but does not affect horn allometry.

Males of the Asian rhinoceros beetle, Trypoxylus dichotomus, possess exaggerated head and thoracic horns that scale dramatically out of proportion to body size. While studies of insulin signaling suggest that this pathway regulates nutrition-dependent growth including exaggerated horns, what regulates disproportionate growth has yet to be identified. The Fat signaling pathway is a potential candidate for regulating disproportionate growth of sexually-selected traits, a hypothesis we advanced in a previous paper (Gotoh et al., 2015). To investigate the role of Fat signaling in the growth and scaling of the sexually dimorphic, condition-dependent traits of the in the Asian rhinoceros beetle T. dichotomus, we used RNA interference to knock down expression of fat and its co-receptor dachsous. Knockdown of fat, and to a lesser degree dachsous, caused shortening and widening of appendages, including the head and thoracic horns. However, scaling of horns to body size was not affected. Our results show that Fat signaling regulates horn growth in T. dichotomus as it does in appendage growth in other insects. However, we provide evidence that Fat signaling does not mediate the disproportionate, positive allometric growth of horns in T. dichotomus.

Females gain survival benefits from immune-boosting ejaculates.

Females in many animal taxa incur significant costs from mating in the form of injury or infection, which can drastically reduce survival. Therefore, immune function during reproduction can be important in determining lifetime fitness. Trade-offs between reproduction and immunity have been extensively studied, yet a growing number of studies demonstrate that mated females have a stronger immune response than virgins. Here, we use the Texas field cricket, Gryllus texensis, to test multiple hypotheses proposed to explain this postmating increase in immune function. Using host-resistance tests, we found that courtship, copulation, and accessory fluids alone do not affect female immunity; rather, only females that acquire intact ejaculates containing testes-derived components exhibit significant increases in survival after exposure to bacterial pathogens. Our data suggest that male-derived components originating from an intact ejaculate and transferred to females during sex are required for the increased immune function characteristic of mated female crickets to occur.

Direct costs and benefits of multiple mating: Are high female mating rates due to ejaculate replenishment?

Females often mate more than is necessary to ensure reproductive success even when they incur significant costs from doing so. Direct benefits are hypothesized to be the driving force of high female mating rates, yet species in which females only receive an ejaculate from their mate still realize increased fitness from multiple mating. Using the Texas field cricket, Gryllus texensis, we experimentally test the hypothesis that multiple mating via monandry or polyandry increases female fitness by replenishing ejaculates, thereby allowing females to produce more offspring for a longer period of time. We found that higher rates of female mating significantly increased lifetime fecundity and oviposition independent of whether females mated with one or two males. Further, although interactions with males significantly increased rates of injury or death, females that replenished ejaculates experienced an increased rate and duration of oviposition, demonstrating that the immediate benefits of multiple mating may greatly outweigh the long-term costs that mating poses to female condition and survival. We suggest that ejaculate replenishment is a driving factor of high mating rates in females that do not receive external direct benefits from mating and that a comparative study across taxa will provide additional insight into the role that ejaculate size plays in the evolution of female mating rates.

Evaluating indices of body condition in two cricket species.

Body mass components (dry mass, lean dry mass, water mass, fat mass) in each sex correlate strongly with body mass and pronotum length in Gryllus texensis and Acheta domesticus. Ordinary least squares (OLS) regression underestimates the scaling relationship between body mass and structural size (i.e., pronotum length) in both cricket species compared with standard major axis (SMA) regression. Standardized mass components correlate more strongly with scaled mass index ([Formula: see text]) than with residual body mass (R i). R i represents the residuals from an OLS regression of log body mass against log pronotum length. Neither condition index predicts energy stores (i.e., fat content) in G. texensis. R i is not correlated with energy stores in A. domesticus whereas [Formula: see text] is negatively correlated. A comparison of condition index methods using published data showed that neither sex nor diet quality affected body condition at adulthood in G. texensis when using the scaled mass index. However, the residual index suggested that sex had a significant effect on body condition. Further, analysis of covariance (ANCOVA) suggested that diet quality significantly affects body mass while statistically controlling for body size (i.e., body condition). We conclude that the statistical assumptions of condition index methods must be met prior to use and urge caution when using methods that are based on least squares in the y -plane (i.e., residual index ANCOVA).