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Proteomic analysis of mature and immature ejaculated spermatozoa from fertile men

Asian Journal of Andrology 2016年第5期18卷 735-746页

作者： Zhihong Cui Rakesh Sharma Ashok Agarwal American Center for Reproductive Medicine Cleveland Clinic Cleveland OH 44195 USA Institute of Toxicology Third Military Medical University Chongqing 400038PR China.

Dysfunctional spermatozoa maturation is the main reason for the decrease in sperm motility and morphology in infertile men. Ejaculated spermatozoa from healthy fertile men were separated into four fractions using three-layer density gradient. Proteins were extracted and bands were digested on a LTQ-Orbitrap Elite hybrid mass spectrometer system. Functional annotations of proteins were obtained using bioinformatics tools and pathway databases. Western blotting was performed to verify the expression levels of the proteins of interest. 1469 proteins were identified in four fractions of spermatozoa. The number of detected proteins decreased according to the maturation level of spermatozoa. During spermatozoa maturation, proteins involved in gamete generation, cell motility, energy metabolism and oxidative phosphorylation processes showed increasing expression levels and those involved in protein biosynthesis, protein transport, protein ubiquitination, and response to oxidative stress processes showed decreasing expression levels. We validated four proteins （HSP 70 1A, clusterin, tektin 2 and tektin 3） by Western blotting. The study shows protein markers that may provide insight into the ejaculated spermatozoa proteins in different stages of sperm maturation that may be altered or modified in infertile men.

关键词： bioinformatics protein validation proteins proteomics spermatozoa maturation

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