Dataset Information

Functional diversity and evolution of the Drosophila sperm proteome

ABSTRACT: Given the central role fertilization plays in the health and fit-ness of sexually reproducing organisms and the well-known evo-lutionary consequences of sexual selection and sperm compe-tition, knowledge gained by a deeper understanding of sperm(and associated reproductive tissues) proteomes has proven crit-5ical to the field’s advancement. Due to their extraordinary complexity, proteome depth-of-coverage is dependent on advance-ments in technology and related bioinformatics, both of which have made significant advancements in the decade since the last Drosophilasperm proteome was published. Here we provide an updated version of the Drosophila melanogastersperm proteome (DmSP3) using improved separation and detection methods andan updated genome annotation. We identified 2563 proteins,with label-free quantitation (LFQ) for 2125 proteins. Combined with previous versions of the sperm proteome, the DmSP3 contains a total of 3176 proteins. The top 20 most abundant pro-teins contained the structural elements α- and β-tubulins and sperm leucyl-aminopeptidases (S-Laps). Both gene content and protein abundance were significantly reduced on the X chromosome, a finding consistent with prior genomic studies of the X chromosome gene content and evolution. We identified 9 of the 16 Y-linked proteins, including known testis-specific male fertility factors. LFQ measured significant levels for 75/83 ribosomal proteins (RPs) we identified, including a number of core constituents. The role of this unique subset of RPs in sperm is unknown. Surprisingly, our expanded sperm proteome also identified 122 seminal fluid proteins (Sfps), proteins found predominantly in the accessory glands. The possibility of tissue contamination from seminal vesicle or other reproductive tissues was addressed using concentrated salt and detergent treatments. Salt treatment had little effect on sperm proteome composition suggesting only minor contamination during sperm isolation while a significant fraction of Sfps remained associate dwith sperm following detergent treatment suggesting Sfps may arise within, and have additional functions in sperm per se.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Drosophila Melanogaster (fruit Fly)

TISSUE(S): Testis, Sperm

SUBMITTER: Tim Karr

LAB HEAD: Tim Karr

PROVIDER: PXD032033 | Pride | 2022-10-14

REPOSITORIES: Pride

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Publications

Functional Diversity and Evolution of the Drosophila Sperm Proteome.

Garlovsky Martin D MD Sandler Jessica A JA Karr Timothy L TL

Molecular & cellular proteomics : MCP 20220817 10

Spermatozoa are central to fertilization and the evolutionary fitness of sexually reproducing organisms. As such, a deeper understanding of sperm proteomes (and associated reproductive tissues) has proven critical to the advancement of the fields of sexual selection and reproductive biology. Due to their extraordinary complexity, proteome depth-of-coverage is dependent on advancements in technology and related bioinformatics, both of which have made significant advancements in the decade since t ...[more]

PMID: 35985624

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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.

Dataset Information

Functional diversity and evolution of the Drosophila sperm proteome

Publications

Functional Diversity and Evolution of the Drosophila Sperm Proteome.

Similar Datasets

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