Journal
ASIAN JOURNAL OF ANDROLOGY
Volume 18, Issue 5, Pages 735-746Publisher
WOLTERS KLUWER MEDKNOW PUBLICATIONS
DOI: 10.4103/1008-682X.164924
Keywords
bioinformatics; protein validation; proteins; proteomics; spermatozoa maturation
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Funding
- American Center for Reproductive Medicine
- NIH shared instrument grant [1S10RR031537-01]
- Chinese Government
- Institute of Toxicology, Third Military Medical University, Chongqing, People's Republic of China
- Lerner Research Institute
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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.
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