Dataset Information

Quantitative proteomics identification of seminal fluid proteins in male Drosophila melanogaster

ABSTRACT: 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.

INSTRUMENT(S): Q Exactive HF, Q Exactive

ORGANISM(S): Drosophila Melanogaster (fruit Fly)

TISSUE(S): Male Reproductive Gland, Ejaculatory Duct, Internal Female Genital Organ, Accessory Sex Gland

SUBMITTER: Philip Charles

LAB HEAD: Stuart Wigby

PROVIDER: PXD009451 | Pride | 2019-08-26

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20170320_MLT_QE752_MSQ988_20170315_BenHopkins_ProG.mgf	Mgf
	20170626_MLT_HF0256_MSQ1071_120617_IremOz_5dil_DvsAGvsBO_ProG.mgf	Mgf
	20170626_MLT_HF0256_MSQ1071_120617_IremOz_5dil_MvsV_ProG2.mgf	Mgf
	F276055.csv	Csv
	F276055.dat	Other

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Publications

Quantitative Proteomics Identification of Seminal Fluid Proteins in Male <i>Drosophila melanogaster</i>.

Sepil Irem I Hopkins Ben R BR Dean Rebecca R Thézénas Marie-Laëtitia ML Charles Philip D PD Konietzny Rebecca R Fischer Roman R Kessler Benedikt M BM Wigby Stuart S

Molecular & cellular proteomics : MCP 20181004 Suppl 1

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 behavior. 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 because of dilution in ...[more]

PMID: 30287546

Publication: 1/2

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