Construction of a proteome profile and functional analysis of the proteins involved in the initiation of mouse spermatogenesis |
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Authors: | Huang Xiao-Yan Guo Xue-Jiang Shen Jian Wang Yu-Feng Chen Lin Xie Jin Wang Ning-Ling Wang Fu-Qiang Zhao Chun Huo Ran Lin Min Wang Xinru Zhou Zuo-Min Sha Jia-Hao |
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Institution: | Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, China. |
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Abstract: | Spermatogenesis is a complex process of terminal differentiation wherein mature sperm are produced. In the first wave of mouse spermatogenesis, different spermatogenic cells appear at specific time points, and their appearance is expected to be accompanied by changes in specific protein expression patterns. In this study, we used 2D-PAGE and MALDI-TOF/TOF technology to construct a comparative proteome profile for mouse testis at specific time points (days 0, 7, 14, 21, 28, and 60 postpartum). We identified 362 differential protein spots corresponding to 257 different proteins. Further cluster analysis revealed 6 expression patterns, and bioinformatics analysis revealed that each pattern was related to many specific cell processes. Among them, 28 novel proteins with unknown functions neither in somatic cells nor germ cells were identified, 8 of which were found to be uniquely or highly expressed in mouse testes via comparison with the GNF SymAtlas database. Further, we randomly selected 7 protein spots and the above 8 novel proteins to verify the expression pattern via Western blotting and RT-PCR, and 6 proteins with little information in testis were further investigated to explore their cellular localization during spermatogenesis by performing immunohistochemistry for the mouse testis tissue. Taken together, the above results reveal an important proteome profile that is functional during the first wave of mouse spermatogenesis, and they provide a strong basis for further research. |
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