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Huang Y.-L.,Guangxi University | Huang Y.-L.,Guangxi High Education Laboratory for Animal Reproduction and Biotechnology | Fu Q.,Guangxi University | Fu Q.,Guangxi High Education Laboratory for Animal Reproduction and Biotechnology | And 11 more authors.
Reproduction in Domestic Animals | Year: 2015

This study was undertaken to investigate differences in protein expression between high- and low-motility sperm of swamp buffalo. The research used two-dimensional gel electrophoresis (2DE) coupled to matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF/TOF-MS) to analyse the different proteins. The results showed 18 different expression protein spots between high- and low-motility buffalo sperm; eight of these proteins were up-regulated in low-motility sperm, five were down-regulated, one deleted and four proteins specifically expressed. Finally, four proteins were successfully identified by MS as belonging to three unique proteins; they are outer dense fibre of sperm tails protein 2 (ODF2), ATP synthase subunit alpha (ATP5A1) and succinyl-CoA synthetase subunit beta (SUCLG2). In summary, these results help to develop an understanding of the molecular mechanisms associated with low-motility sperm and provide clues for finding molecular markers associated with sperm motility. © 2015 Blackwell Verlag GmbH.


Huang Y.-L.,Guangxi University | Huang Y.-L.,Guangxi High Education Laboratory for Animal Reproduction and Biotechnology | Fu Q.,Guangxi University | Fu Q.,Guangxi High Education Laboratory for Animal Reproduction and Biotechnology | And 16 more authors.
Proteomics | Year: 2016

The testicular seminiferous tubules contain Sertoli cells and different types of spermatogenic cells. They provide the microenvironment for spermatogenesis, but the precise molecular mechanism of spermatogenesis is still not well known. Here, we have employed tandem mass tag coupled to LC-MS/MS with the high-throughput quantitative proteomics technology to explore the protein expression from buffalo testicular seminiferous tubules at three different developmental stages (prepuberty, puberty, and postpuberty). The results show 304 differentially expressed proteins with a ≥2-fold change, and bioinformatics analysis indicates that 27 of these may be associated with spermatogenesis. Expression patterns of seven selected proteins were verified via Western blot and quantitative RT-PCR analysis, and further cellular localizations of these proteins by immunohistochemical or immunofluorescence analysis. Taken together, the results provide potential molecular markers of spermatogenesis and provide a rich resource for further studies on male reproduction regulation. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

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