Construction of a proteome profile and functional analysis of the proteins involved in the initiation of mouse spermatogenesis.
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.
Huang
Xiao-Yan
XY
Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, China.
Guo
Xue-Jiang
XJ
Shen
Jian
J
Wang
Yu-Feng
YF
Chen
Lin
L
Xie
Jin
J
Wang
Ning-Ling
NL
Wang
Fu-Qiang
FQ
Zhao
Chun
C
Huo
Ran
R
Lin
Min
M
Wang
Xinru
X
Zhou
Zuo-Min
ZM
Sha
Jia-Hao
JH
D000818:Animals; D015153:Blotting, Western; D016000:Cluster Analysis; D019295:Computational Biology; D015180:Electrophoresis, Gel, Two-Dimensional; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D020543:Proteome / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013091:Spermatogenesis; D013737:Testis / Q000378:metabolism; D013997:Time Factors
D020543:Proteome