Male Seminal Fluid Research Articles

Male condition and seminal fluid affect female host-marking behavior in the Mexican fruit fly

Mating and the transfer of seminal fluid components including male accessory glands (MAGs) proteins can affect oviposition behavior in insects. After oviposition, some species of fruit flies deposit a host-marking pheromone (HMP) on the fruit that discourages oviposition by other females of the same or different species or genus and reduces competition between larvae. However, we know very little about how mating, receiving seminal fluid, or male condition can affect female host marking behavior. Here, we tested how the physiological state of females (mated or unmated), the receipt of seminal fluid, and the condition of the male (wild or sterile) affect oviposition and host-marking behavior (HMB) in Anastrepha ludens (Diptera: Tephritidae). We also determined the efficiency of the host-marking pheromone from mated or unmated females in deterring oviposition. In a further examination of how seminal fluid may be affecting HMB we assessed if there were differences in the size of wild or sterile MAGs and the protein quantity transferred during mating. Our results indicate that receiving seminal fluid increased egg laying and increased time invested in host-marking (HM). Unmated females laid fewer eggs than mated females but invested the same amount of time in depositing host-marking pheromone, which had similar effectiveness in deterring oviposition as that of mated females. Females that mated with sterile males laid the same number of eggs as females that mated with wild males but spent less time depositing host-marking pheromone, which suggests that females detect the condition of the male and invest less in marking hosts. Finally, sterile males had larger accessory glands and transferred more MAGs proteins during mating compared to wild males. Seminal proteins could be manipulating HM behavior and female investment into their current reproductive effort. We are only beginning to understand how male condition and seminal fluid can affect female physiology and maternal investment in HMP.

O-Glycosylation of a male seminal fluid protein influences sperm binding and female postmating behavior.

Glycoproteins are abundant within the human reproductive system and alterations in glycosylation lead to reproductive disorders, suggesting that glycans play an important role in reproductive function. In this study, we used the Drosophila reproductive system as a model to investigate the biological functions of O-glycosylation. We found that O-glycosylation in the male accessory glands, an organ responsible for secreting seminal fluid proteins, plays important roles in female postmating behavior. The loss of one O-glycosyltransferase, PGANT9, in the male reproductive system resulted in decreased egg production in mated females. We identified one substrate of PGANT9, lectin-46Ca (CG1656), which is known to affect female postmating responses. We further show that the loss of lectin-46Ca O-glycosylation affects its ability to associate with sperm tails, resulting in reduced transfer within the female reproductive system. Our results provide the first example that O-glycosylation of a seminal fluid protein affects its ability to associate with sperm in vivo. These studies may shed light on the biological function of O-glycans in mammalian reproduction.

Comparative Genomic Analysis of the Pattern of Evolution of Male and Female Reproductive Proteins in Seed Beetles.

Male seminal fluid proteins often show signs of positive selection and divergent evolution, believed to reflect male-female coevolution. Yet, our understanding of the predicted concerted evolution of seminal fluid proteins and female reproductive proteins is limited. We sequenced, assembled, and annotated the genome of two species of seed beetles allowing a comparative analysis of four closely related species of these herbivorous insects. We compare the general pattern of evolution in genes encoding seminal fluid proteins and female reproductive proteins with those in digestive protein genes and well-conserved reference genes. We found that female reproductive proteins showed an overall ratio of nonsynonymous to synonymous substitutions (ω) similar to that of conserved genes, while seminal fluid proteins and digestive proteins exhibited higher overall ω values. Further, seminal fluid proteins and digestive proteins showed a higher proportion of sites putatively under positive selection, and explicit tests showed no difference in relaxed selection between protein types. Evolutionary rate covariation analyses showed that evolutionary rates among seminal fluid proteins were on average more closely correlated with those in female reproductive proteins than with either digestive or conserved genes. Gene expression showed the expected negative covariation with ω values, except for male-biased genes where this negative relationship was reversed. In conclusion, seminal fluid proteins showed relatively rapid evolution and signs of positive selection. In contrast, female reproductive proteins evolved at a lower rate under selective constraints, on par with genes known to be well conserved. Although our findings provide support for concerted evolution of seminal fluid proteins and female reproductive proteins, they also suggest that these two classes of proteins evolve under partly distinct selective regimes.

Results of HDV RNA detection in male seminal fluid and female cervical swab samples using RT-PCR

Mongolia has an extremely high prevalence of hepatitis D virus (HDV), which leads to increased morbidity and mortality from hepatocellular carcinoma (HCC). HDV is the most severe form of chronic viral hepatitis (CVH). A recent study estimated that the prevalence of anti-HDV was 67.5% among hepatitis B surface antigen-positive cases in Mongolia. HDV infection is particularly prevalent in sexually active young people. The virus may be transmitted through sexual intercourse, and it is crucial to confirm this by detecting HDV RNA in semen and cervical swabs of patients with chronic hepatitis delta. This pilot study was conducted with 16 participants, including eight males and eight females. Viral RNA was isolated from the blood, male seminal fluid, and female cervical swab samples of all participants in the study. HDV RNA was quantified using RT-PCR in samples from blood, seminal fluid, and cervical swabs. This study found that the detection rate of HDV RNA differed between male and female samples, with 62.5% of seminal fluid samples in the male cohort being positive compared with 87.5% in the female cohort. The data shows a statistically significant difference between genders for the presence of HDVRNA in seminal fluid and cervical swabs (OR 0.238, 95% CI= 0.02-3.01). A significant correlation was found between blood HDV RNA quantity and HDV RNA presence status in seminal fluid and cervical swab samples, indicating a potential risk of sexual HDV transmission. The R2 values for semen and cervical swab samples were 0.1095 and 0.9755, respectively.

Reproductive Immunology and Embryo Acceptance

Pregnancy is a test of the fundamental laws of transplantation immunology - the coexistence of genetically disparate tissues without host immune rejection. Advances over recent decades show that implantation and placental development depend on an active state of maternal immune tolerance, and breakdown of tolerance is a key contributor to infertility and recurrent pregnancy loss. Tolerance in pregnancy depends on regulatory T (Treg) cells, which are critical to inhibit effector immunity, constrain inflammation, and support maternal vascular adaptations that allow placental access to maternal nutrient supply. Treg cell defects are implicated in disorders of embryo implantation and placental development, but the origins of Treg cell dysfunction are unknown. In recent studies, we have comprehensively analysed the phenotypes and transcriptional profile of peripheral blood Treg cells in individuals with early pregnancy failure (recurrent miscarriage and recurrent implantation failure). Compared to fertile women, women with early pregnancy failure have fewer total Treg cells and altered Treg cell phenotypes, accompanied by lower Treg:Th1 and Treg:Th17 ratios. RNAseq demonstrates an aberrant Treg cell gene expression profile in early pregnancy failure, with upregulation of pro-inflammatory genes including CSF2, IL4, IL17A, IL21, and IFNG. Our data indicate that Treg cell defects in women with early pregnancy failure can arise due to loss of lineage fidelity associated with impaired FOXP3 regulation. Factors including metabolic function, luteal phase progesterone, and male partner seminal fluid components are all implicated in causing Treg cell deficit. Treg cells may be a useful target for therapeutic interventions to improve uterine receptivity, but this will depend on developing diagnostic tests that are informative. Moreover, safe, and effective interventions to modulate these cells are in their infancy, and personalized approaches matched to specific diagnostic criteria will be needed.

Female reproductive fluid and male seminal fluid: a non-gametic conflict for post-mating control.

Growing evidence shows that non-gametic components released by both males and females can significantly drive sperm competition outcomes. Seminal fluid (SF) was shown to influence paternity success by affecting rival males' sperm performance, and, in some species with male alternative reproductive tactics, to selectively decrease the fertilization success of males of the opposite tactic. Female reproductive fluid (FRF) has been proven to differentially influence ejaculates of different males and bias fertilization towards specific partners. Whether, and with what outcome, these two processes can intersect to influence sperm competition is still unknown. Here we explore this scenario in the grass goby (Zosterisessor ophiocephalus), a fish with territorial-sneaker reproductive tactics, where sneaker males can exploit the territorials' SF while penalizing territorial sperm performance with their own fluid. To test whether FRF can rebalance the ejaculate competition in favour of territorial males, we used in vitro fertilization with a SF mixture (territorial + sneaker), using increasing concentrations of FRF, to simulate the natural conditions that ejaculates encounter towards the eggs. Our findings revealed a differential effect of FRF on the different tactics' fertilization success, favouring territorial ejaculates, possibly through an attenuation of the detrimental effects of sneaker SF, and enabling females to regain control over the fertilization process.

WMel Wolbachia alters female post-mating behaviors and physiology in the dengue vector mosquito Aedes aegypti

Globally invasive Aedes aegypti disseminate numerous arboviruses that impact human health. One promising method to control Ae. aegypti populations is transinfection with Wolbachia pipientis, which naturally infects ~40–52% of insects but not Ae. aegypti. Transinfection of Ae. aegypti with the wMel Wolbachia strain induces cytoplasmic incompatibility (CI), allows infected individuals to invade native populations, and inhibits transmission of medically relevant arboviruses by females. Female insects undergo post-mating physiological and behavioral changes—referred to as the female post-mating response (PMR)—required for optimal fertility. PMRs are typically elicited by male seminal fluid proteins (SFPs) transferred with sperm during mating but can be modified by other factors, including microbiome composition. Wolbachia has modest effects on Ae. aegypti fertility, but its influence on other PMRs is unknown. Here, we show that Wolbachia influences female fecundity, fertility, and re-mating incidence and significantly extends the longevity of virgin females. Using proteomic methods to examine the seminal proteome of infected males, we found that Wolbachia moderately affects SFP composition. However, we identified 125 paternally transferred Wolbachia proteins, but the CI factor proteins (Cifs) were not among them. Our findings indicate that Wolbachia infection of Ae. aegypti alters female PMRs, potentially influencing control programs that utilize Wolbachia-infected individuals.

Paternity Test

The article presents modern data on the properties of seminal fluid creating the primary conditions for the development of a healthy pregnancy. Actual pathophysiological theories based on the study of cell lines, proving the direct role of paternal material in the formation of an allogeneic environment, are considered. Literature data on the influence of dosing and quality of the partner's biological material are given. The subject of fertility is the interaction between male seminal fluid and female tissues during sexual intercourse, the effect on fertility, pregnancy and the health of the child. It has been established that the biological contribution of the father to pregnancy and his ability to influence the reproductive outcome goes beyond the simple provision of male mi gametes at conception. The key questions of the study are: whether the seminal fluid delivered during sexual intercourse affects gene expression and immune responses to the endometrium; the completeness of the identity of the molecular agents by which seminal plasma and sperm interact with the cells of the cervix and endometrium; the physiological significance of semen priming in women and its potential contribution to unexplained infertility, embryonic development and pregnancy outcome; clinical factors, lifestyle and environmental environments that affect seminal fluid composition and signaling function. Ultimately, a better understanding of the role of seminal fluid in shaping reproductive success could provide new avenues for diagnosing and treating infertility in couples and improve recommendations for couples planning a pregnancy or in assisted reproduction settings.

Evolutionary Quantitative Proteomics of Reproductive Protein Divergence in Drosophila

Reproductive traits often evolve rapidly between species. Understanding the causes and consequences of this rapid divergence requires characterization of female and male reproductive proteins and their effect on fertilization success. Species in the Drosophila virilis clade exhibit rampant interspecific reproductive incompatibilities, making them ideal for studies on diversification of reproductive proteins and their role in speciation. Importantly, the role of intraejaculate protein abundance and allocation in interspecific divergence is poorly understood. Here, we identify and quantify the transferred male ejaculate proteome using multiplexed isobaric labeling of the lower female reproductive tract before and immediately after mating using three species of the virilis group. We identified over 200 putative male ejaculate proteins, many of which show differential abundance between species, suggesting that males transfer a species-specific allocation of seminal fluid proteins during copulation. We also identified over 2000 female reproductive proteins, which contain female-specific serine-type endopeptidases that showed differential abundance between species and elevated rates of molecular evolution, similar to that of some male seminal fluid proteins. Our findings suggest that reproductive protein divergence can also manifest in terms of species-specific protein abundance patterns.

The Effects of Male Seminal Fluid Proteins on Gut/Gonad Interactions in Drosophila.

Mating initiates broad physiological changes encompassing multiple organ systems in females. Elucidating the complex inter- and intra-organ signaling events that coordinate these physiological changes is an important goal in the field of reproductive biology. Further characterization of these complex molecular and physiological interactions is key to understanding how females meet the energetic demands of offspring production. Many recent studies of the fruit fly, Drosophila melanogaster, have described the mechanisms of post-mating changes within the female reproductive tract and digestive system. Additionally, other studies have described post-mating signaling crosstalk between these systems. Interestingly, male seminal fluid proteins have been linked to post-mating responses within the female reproductive tract and gut, and to signaling events between the two organ systems. However, information about the hormonal and neuronal signaling pathways underlying the post-mating signaling events within and between the reproductive tract and digestive systems that are triggered by seminal fluid proteins has yet to be combined into a single view. In this article, we summarize and integrate these studies into a single "network schematic" of the known signaling events within and between the reproductive and digestive systems downstream of male seminal fluid proteins. This synthesis also draws attention to the incomplete parts of these pathways, so that outstanding questions may be addressed in future studies.

A single mating is sufficient to induce persistent reduction of immune defense in mated female Drosophila melanogaster

In many species, female reproductive investment comes at a cost to immunity and resistance to infection. Mated Drosophila melanogaster females are more susceptible to bacterial infection than unmated females. Transfer of the male seminal fluid protein Sex Peptide reduces female post-mating immune defense. Sex Peptide is known to cause both short- and long-term changes to female physiology and behavior. While previous studies showed that females were less resistant to bacterial infection as soon as 2.5 h and as long as 26.5 h after mating, it is unknown whether this is a binary switch from mated to unmated state or whether females can recover to unmated levels of immunity. It is additionally unknown whether repeated mating causes progressive reduction in defense capacity. We compared the immune defense of mated females when infected at 2, 4, 7, or 10 days after mating to that of unmated females and saw no recovery of immune capacity regardless of the length of time between mating and infection. Because D. melanogaster females can mate multiply, we additionally tested whether a second mating, and therefore a second transfer of seminal fluids, caused deeper reduction in immune performance. We found that females mated either once or twice before infection survived at equal proportions, both with significantly lower probability than unmated females. We conclude that a single mating event is sufficient to persistently suppress the female immune system. Interestingly, we observed that induced levels of expression of genes encoding antimicrobial peptides (AMPs) decreased with age in both experiments, partially obscuring the effects of mating. Collectively, the data indicate that being reproductively active versus reproductively inactive are alternative binary states with respect to female D. melanogaster immunity. The establishment of a suppressed immune status in reproductively active females can inform our understanding of the regulation of immune defense and the mechanisms of physiological trade-offs.

Functional morphology of the seminal receptacle of Mithrax, Mithraculus and Omalacantha spider crabs (Brachyura: Mithracidae)

Most Majoidea crabs display high sperm competition rates due to the development of spermatic strata within the seminal receptacle (SR). To verify the occurrence of sperm competition, SR anatomy and histochemistry analyses were performed in Mithrax hispidus, Omalacantha bicornuta, and Mithraculus forceps. The SR of the three Mithracidae species is classified into two regions, one of mesodermal origin (dorsal), consisting of a stratified epithelium with desquamation cells that produce a holocrine secretion, and the other, an ectodermal region (ventral) comprising a simple cubic epithelium covered by a cuticle. The oviduct opens in a ventral position near the transition region, which exhibits more pronounced folds on the opposite face of the oviduct, which may aid fertilization due to the presence of an external musculature. Sperm masses in a circular format were observed in the SR lumen, reminiscent of coenospermic spermatophores, and no morphological evidence strata of sperm packets were observed in any of the three studied species. The ventral SR followed the most common pattern observed in Majoidea. The secretion produced in the receptacle is composed of glycoproteins, with neutral polysaccharides. Acidic polysaccharides probably play a role against microorganisms from male seminal fluid. Due to the absence sperm packets, we were unable to determine whether the investigated females receive material from more than one male nor (if this does, in fact, occur) whether any preference for the spermatozoa of one male over another takes place during fertilization, that can may indicate the absence of sperm competition in the investigated species.

Drosophila female reproductive glands contribute to mating plug composition and the timing of sperm ejection.

Reproductive traits that influence female remating and competitive fertilization rapidly evolve in response to sexual selection and sexual conflict. One such trait, observed across diverse animal taxa, is the formation of a structural plug inside the female reproductive tract (FRT), either during or shortly after mating. In Drosophila melanogaster, male seminal fluid forms a mating plug inside the female bursa, which has been demonstrated to influence sperm entry into storage and latency of female remating. Processing of the plug, including its eventual ejection from the female's reproductive tract, influences the competitive fertilization success of her mates and is mediated by female × male genotypic interactions. However, female contributions to plug formation and processing have received limited attention. Using developmental mutants that lack glandular FRT tissues, we reveal that these tissues are essential for mating plug ejection. We further use proteomics to demonstrate that female glandular proteins, and especially proteolytic enzymes, contribute to mating plug composition and have a widespread impact on plug formation and composition. Together, these phenotypic and molecular data identify female contributions to intersexual interactions that are a potential mechanism of post-copulatory sexual selection.

Could Sterile Aedes albopictus Male Releases Interfere with Aedes aegypti Population in Reunion Island?

Simple SummaryThe Sterile Insect Technique consisting of inundative and repeated releases of sterile males that induce sterility in the wild population is currently tested as a new strategy to control Ae. albopictus. Considering that Ae. albopictus and Ae. aegypti thrive in sympatry in some regions, and that mating between both species is known to occur at low rates, we hypothesize that releasing millions of sterile male Ae. albopictus in could affect female Ae. aegypti reproduction. To study this potential mating interference, the newly established marking technique has been applied using the rhodamine B that is administered to males through sugar meal. It is internally incorporated into the mosquito’s male body and during mating, the rhodamine is transferred into the females. In laboratory-based experiments rhodamine marking proved to be a powerful means of detecting mating in females of both Aedes species, whatever the mating crosses between males and females. Some mated females were able to lay eggs, but all were not viable. However, despite the promiscuity of the adults in small experimental cages, 95% of the female Ae aegypti showed no evidence of insemination following mating with sterile male Ae. albopictus, suggesting that in the field, an inundative and repeated releases of sterile male Ae. albopictus will not influence the reproduction of female Ae aegypti.In Reunion Island, the feasibility of an Aedes albopictus control program using the Sterile Insect Technique (SIT) is studied. Because, in some regions, Ae. albopictus is living in sympatry with Aedes aegypti, the impact of releasing millions of sterile male Ae. albopictus on female Ae. aegypti reproduction needs to be assessed. Thus, to study the potential heterospecific matings, a marking technique using rhodamine B has been used. Rhodamine is given in solution to male mosquitoes to be incorporated into the male body and seminal fluid and transferred during mating into the bursa inseminalis and spermathecae of females. The presence of rhodamine in females occurred in 15% of cases when Ae. aegypti females were offered non-irradiated Ae. albopictus males, 5% when offered irradiated Ae. albopictus males and 18% of cases in the inverse heterospecific matings. Moreover, our results also showed that these matings gave few eggs but were not viable. Finally, the results showed that whatever the type of mating crosses, females in cages previously crossed with males of another species can re-mate with males of their species and produce an equivalent amount of egg compared to females only mated with conspecific males. Despite the promiscuity of the males and females in small cages for three days, heterospecific mating between sterile male Ae. albopictus and female Ae aegypti, 95% of the females have not been inseminated suggesting that in the field the frequency satyrization would be very low.

Regulation of Mating-Induced Increase in Female Germline Stem Cells in the Fruit Fly Drosophila melanogaster

In many insect species, mating stimuli can lead to changes in various behavioral and physiological responses, including feeding, mating refusal, egg-laying behavior, energy demand, and organ remodeling, which are collectively known as the post-mating response. Recently, an increase in germline stem cells (GSCs) has been identified as a new post-mating response in both males and females of the fruit fly, Drosophila melanogaster. We have extensively studied mating-induced increase in female GSCs of D. melanogaster at the molecular, cellular, and systemic levels. After mating, the male seminal fluid peptide [e.g. sex peptide (SP)] is transferred to the female uterus. This is followed by binding to the sex peptide receptor (SPR), which evokes post-mating responses, including increase in number of female GSCs. Downstream of SP-SPR signaling, the following three hormones and neurotransmitters have been found to act on female GSC niche cells to regulate mating-induced increase in female GSCs: (1) neuropeptide F, a peptide hormone produced in enteroendocrine cells; (2) octopamine, a monoaminergic neurotransmitter synthesized in ovary-projecting neurons; and (3) ecdysone, a steroid hormone produced in ovarian follicular cells. These humoral factors are secreted from each organ and are received by ovarian somatic cells and regulate the strength of niche signaling in female GSCs. This review provides an overview of the latest findings on the inter-organ relationship to regulate mating-induced female GSC increase in D. melanogaster as a model. We also discuss the remaining issues that should be addressed in the future.

Temporal and sequential order of nonoverlapping gene networks unraveled in mated female Drosophila.

In this study, we reanalyzed available datasets of gene expression changes in female Drosophila head induced by mating. Mated females present metabolic phenotypic changes and display behavioral characteristics that are not observed in virgin females, such as repulsion to male sexual aggressiveness, fidelity to food spots selected for oviposition, and restriction to the colonization of new niches. We characterize gene networks that play a role in female brain plasticity after mating using AMINE, a novel algorithm to find dysregulated modules of interacting genes. The uncovered networks of altered genes revealed a strong specificity for each successive period of life span after mating in the female head, with little conservation between them. This finding highlights a temporal order of recruitment of waves of interconnected genes which are apparently transiently modified: the first wave disappears before the emergence of the second wave in a reversible manner and ends with few consolidated gene expression changes at day 20. This analysis might document an extended field of a programmatic control of female phenotypic traits by male seminal fluid.

Toll-like receptor-4 null mutation causes fetal loss and fetal growth restriction associated with impaired maternal immune tolerance in mice

Maternal immune adaptation to accommodate pregnancy depends on sufficient availability of regulatory T (Treg) cells to enable embryo implantation. Toll-like receptor 4 is implicated as a key upstream driver of a controlled inflammatory response, elicited by signals in male partner seminal fluid, to initiate expansion of the maternal Treg cell pool after mating. Here, we report that mice with null mutation in Tlr4 (Tlr4−/−) exhibit impaired reproductive outcomes after allogeneic mating, with reduced pregnancy rate, elevated mid-gestation fetal loss, and fetal growth restriction, compared to Tlr4+/+ wild-type controls. To investigate the effects of TLR4 deficiency on early events of maternal immune adaptation, TLR4-regulated cytokines and immune regulatory microRNAs were measured in the uterus at 8 h post-mating by qPCR, and Treg cells in uterus-draining lymph nodes were evaluated by flow cytometry on day 3.5 post-coitum. Ptgs2 encoding prostaglandin-endoperoxide synthase 2, cytokines Csf2, Il6, Lif, and Tnf, chemokines Ccl2, Cxcl1, Cxcl2, and Cxcl10, and microRNAs miR-155, miR-146a, and miR-223 were induced by mating in wild-type mice, but not, or to a lesser extent, in Tlr4−/− mice. CD4+ T cells were expanded after mating in Tlr4+/+ but not Tlr4−/− mice, with failure to expand peripheral CD25+FOXP3+ NRP1− or thymic CD25+FOXP3+ NRP1+ Treg cell populations, and fewer Treg cells expressed Ki67 proliferation marker and suppressive function marker CTLA4. We conclude that TLR4 is an essential mediator of the inflammation-like response in the pre-implantation uterus that induces generation of Treg cells to support robust pregnancy tolerance and ensure optimal fetal growth and survival.

Interspecific Mating Effects on Locomotor Activity Rhythms and Refractoriness of Aedes albopictus (Diptera: Culicidae) Females.

Simple SummaryThe superiority of Aedes albopictus in larval resource competition was originally proposed as the cause of displacements of Aedes aegypti in the USA. However, satyrization, a form of reproductive interference, was later invoked as an alternative or complementary mechanism for the observed displacements of Aedes aegypti. This study tests the hypotheses that Ae. albopictus female activity is not altered by the presence of accessory gland extracts from conspecific and heterospecific males, and Ae. albopictus females remain receptive to conspecific males even after receiving accessory gland (AG) extracts from Ae. aegypti males. We performed experiments with a control group (females injected with saline), a group of females injected with accessory gland extracts of Ae. aegypti males and a group of females injected with accessory gland extracts of Ae. albopictus males and measured the locomotor activity and the ability of inseminated females to copulate with conspecific males. Females injected with conspecific and heterospecific extracts showed significant decreases in total and diurnal activity. Females injected with heterospecific extracts showed significant decreases in nocturnal activity. A total of 83% of females injected with heterospecific and 10% of females injected with conspecific extracts copulated with conspecific males. These results considered together with our previous report on effects of interspecific mating and cross-species injections of AG products on Ae. aegypti females, show consistent depressions of locomotor activities between species, but the loss of sexual receptivity only in Ae. aegypti. We propose that different male seminal fluid proteins control these activities.This study tests the hypotheses that the locomotor activity of Ae. albopictus females is not significantly altered by the presence of accessory gland (AG) extracts from conspecific and heterospecific males, and that Ae. albopictus females remain receptive to mating with conspecific males even after receiving AG of Ae. aegypti males. Virgin Ae. albopictus females were injected with saline (control group), AG extracts of Ae. aegypti males (aegMAG) or AG extracts of Ae. albopictus males (albMAG). Locomotor activity was evaluated under 12 h of light and 12 h of darkness at 25 °C. All live Ae. albopictus females were subsequently exposed to conspecific males for 48 h, and their spermathecae were dissected for the presence of sperm. Females injected with aegMAG and albMAG showed significant decreases in total, diurnal and diurnal without lights-on Period activities. Females injected with aegMAG showed significant decreases in nocturnal and nocturnal without lights-off period activities. Females injected with albMAG showed significant decreases in lights-off activity. A total of 83% of Ae. albopictus females injected with aegMAG and 10% of females injected with albMAG were inseminated by conspecific males. These results, coupled with our previous paper on MAG and interspecific mating effects on female Ae. aegypti, demonstrate contrasting outcomes on locomotor activities and loss of sexual receptivity, both conspecific and heterospecific MAGs capable of sterilizing virgin Ae. aegypti, but only conspecific MAGs sterilizing Ae. albopictus, whereas locomotor activities were depressed in females of both species after heterospecific and conspecific injections or treatments.

Identification of seminal proteins related to the inhibition of mate searching in female crickets

AbstractIn response to the reduction in fitness associated with sperm competition, males are expected to evolve tactics that hinder female remating. For example, females often display a postmating reduction in their sexual receptivity that has been shown to be mediated by proteins contained in a male’s seminal fluid (sfps). However, although there has been comprehensive research on sfps in genetically well-characterized species, few nonmodel species have been studied in such detail. We initially confirm that female Australian field crickets, Teleogryllus oceanicus, do display a significant reduction in their mate-searching behavior 24 h after mating. This effect was still apparent 3 days after mating but was entirely absent after 1 week. We then attempted to identify the sfps that might play a role in inducing this behavioral response. We identified two proteins, ToSfp022 and ToSfp011, that were associated with the alteration in female postmating behavior. The knockdown of both proteins resulted in mated females that displayed a significant increase in their mate-searching behaviors compared with females mated to males having the full compliment of seminal fluid proteins in their ejaculate. Our results indicate that the female refractory period in T. oceanicus likely reflects a sperm competition avoidance tactic by males, achieved through the action of male seminal fluid proteins.

Copulation Song in Drosophila: Do Females Sing to Change Male Ejaculate Allocation and Incite Postcopulatory Mate Choice?

Drosophila males sing a courtship song to achieve copulations with females. Females were recently found to sing a distinct song during copulation, which depends on male seminal fluid transfer and delays female remating. Here, it is hypothesized that female copulation song is a signal directed at the copulating male and changes ejaculate allocation. This may alter female remating and sperm usage, and thereby affect postcopulatory mate choice. Mechanisms of how female copulation song is elicited, how males respond to copulation song, and how remating is modulated, are considered. The potential adaptive value of female signaling during copulation is discussed with reference to vertebrate copulation calls and their proposed function in eliciting mate guarding. Female copulation song may be widespread within the Drosophila genus. This newly discovered behavior opens many interesting avenues for future research, including investigation of how sexually dimorphic neuronal circuits mediate communication between nervous system and reproductive organs.