Dataset Information

Quantitative proteomic analysis of seminal plasma, sperm membrane proteins and seminal extracellular vesicles suggests vesicular mechanisms aid in the removal and addition of proteins to the ram sperm membrane.

ABSTRACT: Quantitative proteomic studies are contributing greatly to our understanding of the spermatozoon through the provision of detailed information on the proteins spermatozoa acquire and shed in the acquisition of fertility. Extracellular vesicles (EVs) are thought to aid in the delivery of proteins to spermatozoa in the male reproductive tract. The aim of this study was to identify EV proteins isolated from ram seminal plasma. High-speed centrifugation using Optiprep sucrose gradients was used to purify two EV populations from seminal plasma. Liquid chromatography and tandem mass spectrometry (LC-MS/MS) with quantitative SWATH was used to identify proteins enriched in the extracellular vesicle preparation (e.g. EDIL3) and those that were co-isolated with vesicular preparations due to their abundance or adhesive nature (e.g. BSPs). Ram sperm plasma membrane proteins were also isolated using nitrogen cavitation and identified with LC-MS/MS to better understand the interplay of proteins between the sperm membrane and extracellular environment. The categorisation of proteins enriched in the EV population (P<0.05, 2-fold increase compared to whole seminal plasma) according to their function revealed three main groupings: vesicle biogenesis, metabolism and membrane adhesion and remodelling. The latter group contained many reproduction-specific proteins that show demonstrable links to sperm fertility. Many of these membrane-bound proteins show testicular expression and are shed from the sperm surface during epididymal maturation (e.g. TEX101, LY6K). Their association with seminal EVs suggests that EVs may not only deliver protein cargo to spermatozoa but also assist in the removal of proteins from the sperm membrane.

INSTRUMENT(S): TripleTOF 6600

ORGANISM(S): Ovis Aries

TISSUE(S): Spermatozoon, Sperm, Seminal Plasma

SUBMITTER: Taylor Pini

LAB HEAD: Taylor Pini

PROVIDER: PXD017537 | Pride | 2020-05-08

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20170404_JR_SWATH_B1_rep1.wiff.zip	Wiff
	20170404_JR_SWATH_B1_rep1.zip	Other
	20170404_JR_SWATH_B1_rep2.wiff.zip	Wiff
	20170404_JR_SWATH_B1_rep2.zip	Other
	20170404_JR_SWATH_B2_rep1.wiff.zip	Wiff

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Quantitative Proteomic Analysis of Seminal Plasma, Sperm Membrane Proteins, and Seminal Extracellular Vesicles Suggests Vesicular Mechanisms Aid in the Removal and Addition of Proteins to the Ram Sperm Membrane.

Leahy Tamara T Rickard Jessica P JP Pini Taylor T Gadella Bart M BM de Graaf Simon P SP

Proteomics 20200608 12

Quantitative proteomic studies are contributing greatly to the understanding of the spermatozoon through the provision of detailed information on the proteins spermatozoa acquire and shed in the acquisition of fertility. Extracellular vesicles (EVs) are thought to aid in the delivery of proteins to spermatozoa in the male reproductive tract. The aim of this study is to isolate, identify and quantify EV proteins isolated from ram seminal plasma. Ram sperm plasma membrane proteins are also isolate ...[more]

PMID: 32383290

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