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Effect of receipt of male ejaculate sex peptide on micro RNA expression in female Drosophila melanogaster, 3 hours after the start of mating, in HeadThorax (HT) and Abdomen (Abd) tissues.

ABSTRACT: Seminal fluid proteins (Sfps) can have significant effects on female behaviour and physiology. In Drosophila melanogaster a single Sfp, sex peptide (SP), triggers a remarkably diverse array of fundamentally important responses. It increases female fecundity, juvenile hormone (JH) production, antimicrobial peptide production and feeding rates, whilst decreasing sexual receptivity. Although these features are expected to leave a strong genomic signature, previous studies indicated that Sfps affect the expression of relatively modest numbers of genes. A potential explanation is that increased resolution is needed or that SP could also elicit post-transcriptional control at the level of microRNAs (miRs). We hypothesised that responses to SP at both the transcriptional and post-transcriptional levels could provide precise temporal and spatial responsiveness and facilitate the expression of diverse phenotypes from a single ligand. To test this idea, we used microarrays and Solexa deep sequencing to profile mRNA and miR responses to SP in adult female abdomen (Abd) and head+thorax (HT) tissues. SP elicited a rich but subtle signature of temporally and spatially controlled mRNAs related to functions that match the predicted known effects of SP, but that also suggest potentially novel roles for SP such as in phototransduction. mRNA profiles were much more varied in the HT than in the Abd, both within and across time points (3 and 6h after mating). The deep sequencing data revealed post-transcriptional control of SP responses by differential expression of miRs in adults. As for the mRNA profiles, there was evidence for tissue specificity. miRs showing largest expression differences tended to be down regulated in the HT and up regulated in the Abd. Receipt of SP significantly down-regulated the expression of miR279, a micro-RNA with potentially diverse sensory and reproductive roles, across the HT and Abd. We also identified potential differential expression for miRs 31A, 184, 276A and 989. Our study provides the first evidence of changes in both transcriptional and post-transcriptional expression in adults in response to Sfps, and provides a potential mechanism for the extraordinary diversity of responses seen in response to the single SP stimulus.

ORGANISM(S): Drosophila melanogaster

PROVIDER: GSE31250 | GEO | 2017/08/01

SECONDARY ACCESSION(S): PRJNA146061

REPOSITORIES: GEO

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Project description:Seminal fluid contains some of the fastest evolving proteins currently known. These seminal fluid proteins (Sfps) play crucial roles in reproduction, such as supporting sperm function, and – particularly in insects – modifying female physiology and behaviour. Identification of Sfps in small animals is challenging, and often relies on samples taken from the female reproductive tract after mating. A key pitfall of this method is that it might miss Sfps that are of low abundance due to dilution in the female-derived sample or rapid processing in females. Here we present a new and complimentary method, which provides added sensitivity to Sfp identification. We applied label-free quantitative proteomics to Drosophila melanogaster male reproductive tissue – where Sfps are unprocessed, and highly abundant – and quantified Sfps before and immediately after mating, to infer those transferred during copulation. We also analysed female reproductive tracts immediately before and after copulation to confirm the presence and abundance of known and candidate Sfps, where possible. Results were cross-referenced with transcriptomic and sequence databases to improve confidence in Sfp detection. Our data was consistent with 124 previously reported Sfps. We found 8 high-confidence novel candidate Sfps, which were both depleted in mated versus unmated males and identified within the reproductive tract of mated but not virgin females. We also identified 31 more candidates that are likely Sfps based on their abundance, known expression and predicted characteristics, and revealed that four proteins previously identified as Sfps are at best minor contributors to the ejaculate. The estimated copy numbers for our candidate Sfps were lower than for previously identified Sfps, supporting the idea that our technique provides a deeper analysis of the Sfp proteome than previous studies. Our results demonstrate a novel, high-sensitivity approach to the analysis of seminal fluid proteomes, whose application will further our understanding of reproductive biology.

Project description:Globally invasive Aedes aegypti mosquitoes disseminate numerous arboviruses that impact human health. One promising method to control Ae. aegypti populations is transinfection with the intracellular bacterium Wolbachia pipientis, a symbiont that naturally infects ~40-52% of insects but is normally absent from Ae. aegypti. Transinfection of Ae. aegypti with the wMel Wolbachia strain induces cytoplasmic incompatibility (CI), allowing infected individuals to rapidly invade native populations. Further, wMel Wolbachia-infected females display refractoriness to medically relevant arboviruses. Thus, wMel Wolbachia-infected Ae. aegypti are being released in several areas to replace native populations, thereby suppressing disease transmission by this species. Wolbachia is reported to have minimal effects on Ae. aegypti fertility, but its influence on other reproductive processes is unknown. Female insects undergo several post-mating physiological and behavioral changes required for optimal fertility. Post-mating responses (PMRs) in female insects are typically elicited by receipt of male seminal fluid proteins (SFPs) transferred with sperm during mating, but can be modified by other factors, such as adult age, nutritional status, and microbiome composition. To assess how Wolbachia infection influences Ae. aegypti female PMRs, we collected wMel Wolbachia-infected Ae. aegypti from the field in Medellín, Colombia and introduced the bacterium into our laboratory strain. We found that Wolbachia influences female fecundity, fertility, and re-mating incidence. Further, we observed that Wolbachia significantly extends longevity of virgin females. Changes in female PMRs are not due to defects in sperm transfer by infected males, or sperm storage by infected females. Using proteomic methods to examine the seminal proteome of infected males, we found that Wolbachia infection has a moderate effect on SFP composition. However, we identified 125 Wolbachia proteins that are paternally transferred to females by infected males. Surprisingly, the CI factor proteins (Cifs), were not detected in the ejaculates of Wolbachia-infected males. Our findings indicate that Wolbachia infection of Ae. aegypti alters female post-mating responses, potentially influencing control programs that utilize Wolbachia-infected individuals.

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Effect of receipt of male ejaculate sex peptide on micro RNA expression in female Drosophila melanogaster, 3 hours after the start of mating, in HeadThorax (HT) and Abdomen (Abd) tissues.

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