Drosophila melanogaster
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1099684 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR28737491_1.fastq.gz | Fastqsanger.gz | |||
| SRR28737491_2.fastq.gz | Fastqsanger.gz | |||
| SRR28737492_1.fastq.gz | Fastqsanger.gz | |||
| SRR28737492_2.fastq.gz | Fastqsanger.gz |
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Project description:We present data using a novel method to simultaneously identify and quantify transferred male seminal proteins and the female reproductive proteome using multiplexed Tandem-Mass-Tag (TMT) isobaric labelling of the lower female reproductive tracts dissected from virgin- or recently mated- females of three species of the virilis group. We identified over 200 putative male ejaculate proteins many of which show differential abundance between species. We also identified over 2000 proteins providing the first description of the Drosophila female reproductive tract proteome outside of the melanogaster group which also shows significant divergence between species. We then assessed the utility of species-specific compared to single species query databases for protein identification and quantification.
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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.
2019-08-26 | PXD009451 | Pride
Project description:Spatiotemporal female reproductive tract gene expression in Drosophila melanogaster
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Project description:Fertility depends on the progression of complex and coordinated postmating processes within the extracellular luminal environment of the female reproductive tract (FRT). To achieve a more comprehensive level of knowledge regarding female-derived proteins available to interact with the ejaculate, we utilized semiquantitative mass spectrometry-based proteomics to study the composition of the FRT tissue and, separately, the luminal fluid, before and after mating in Drosophila melanogaster. Our approach leveraged whole-fly isotopic labelling to delineate between female proteins and those transferred from males in the ejaculate. The dynamic mating-induced proteomic changes in the extracellular FRT luminal fluid further informs our understanding of secretory mechanisms of the FRT and serves as a foundation for establishing the roles of ejaculate-female interactions in fertility.
2021-09-22 | PXD025085 | Pride
Project description:Reproductive status alters transcriptomic response to infection in female Drosophila melanogaster
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Project description: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, 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 female reproductive tract tissues, we reveal that these glandular tissues are essential for the mating plug to be ejected. We further use proteomics to demonstrate that female glandular proteins, and especially proteolytic enzymes, contribute to mating plug composition and that the absence of glands has a widespread impact of plug formation and composition. Together, these phenotypic and molecular data resolve molecular mechanisms of important postmating, intersexual interactions and cryptic female choice.
2022-01-25 | PXD028524 | Pride
Project description:Fertility depends on the coordination of complex interactions between male and female reproductive proteins inside the female reproductive tract (FRT). These interactions mediate a suite of changes in female behavior, morphology, and physiology after mating, yet little is known about how the molecular environment of the FRT may differ among species and coordinate species-specific female post-mating responses. We used semi-quantitative proteomics to compare the FRT protein composition between virgin and mated females in Drosophila melanogaster. These results are compared to those from quantitative TMT proteomic analyses of the mating-induced changes in D. simulans and D. mauritiana, and after conspecific and heterospecific inseminations. Our study highlights the value of using quantitative proteomics approaches to study the molecular composition of the FRT environment, and how its divergence may inform mechanistic studies of post-mating pre-zygotic reproductive isolation between species.
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Project description:Transcriptional profiling of 3 day old virgin male and female adults comparing control male Drosophila melanogaster (MDM) versus male D sechellia (MDS) and comparing control female Drosophila melanogaster (FDM) versus female D sechellia (FDS). Goal was to determine why D sechellia is tolerant to octanoïc acid, the major toxic compound of Morinda citrifolia fruit
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Project description:Male-derived accessory gland proteins (Acps) that are transferred to females during mating have profound effects on female reproductive physiology including increased ovulation, mating inhibition, and effects on sperm utilization and storage. The extreme rates of evolution seen in Acps may be driven by sperm competition and sexual conflict, processes which may ultimately drive complex interactions between female- and male-derived molecules and sperm. However, little is known of how gene expression in female reproductive tissues changes in response to the presence of male molecules and sperm. To characterize this response, we conducted parallel genomic and proteomic analyses of gene expression in the reproductive tract of 3-day-old unmated and mated female Drosophila melanogaster. Using DNA microarrays, we identified 539 transcripts that are differentially expressed in unmated vs. mated females and revealed a striking peak in differential expression at 6 hrs postmating and a marked shift from primarily down-regulated to primarily up-regulated transcripts within 3 hrs after mating. Combining two-dimensional gel electrophoresis and liquid chromatography mass spectrometry analyses, we identified 84 differentially expressed proteins at 3 hrs postmating, including proteins which appeared to undergo post-translational modification. Together, our observations define transcriptional and translational response to mating within the female reproductive tract and suggest a bimodal model of postmating gene expression initially correlated with mating and the final stages of female reproductive tract maturation and later with the declining presence of male reproductive molecules and with sperm maintenance and utilization. Experiment Overall Design: Three-day-old mated and unmated females were dissected to remove the lower reproductive tract (upper uterus, sperm-storage organs, and accessory glands). Mated females were dissected either immediately following mating (0 hr) or at 3, 6, or 24 hrs following the termination of mating. Tracts of 12-40 females of like category were pooled and total RNA extracted via a TRIzol-based protocol. Processing and labeling of transcript was performed by the Molecular Biology Core Facility at the Medical College of Georgia. Arrays from mated females at the different timepoints were compared to unmated females.
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