Open AccessEstablishment of a proteome profile and identification of molecular markers for mouse spermatogonial stem cells
Author(s) -
Zhou Quan,
Guo Yueshuai,
Zheng Bo,
Shao Binbin,
Jiang Min,
Wang Gaigai,
Zhou Tao,
Wang Lei,
Zhou Zuomin,
Guo Xuejiang,
Huang Xiaoyan
Publication year - 2015
Publication title -
journal of cellular and molecular medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.44
H-Index - 130
eISSN - 1582-4934
pISSN - 1582-1838
DOI - 10.1111/jcmm.12407
Subject(s) - biology , proteome , microbiology and biotechnology , proteomics , stem cell , transplantation , immunofluorescence , gene , genetics , antibody , medicine , surgery
Spermatogonial stem cells ( SSC s) are undifferentiated cells that are required to maintain spermatogenesis throughout the reproductive life of mammals. Although SSC transplantation and culture provide a powerful tool to identify the mechanisms regulating SSC function, the precise signalling mechanisms governing SSC self‐renewal and specific surface markers for purifying SSC s remain to be clearly determined. In the present study, we established a steady SSC culture according to the method described by Shinohara's lab. Fertile progeny was produced after transplantation of cultured SSC s into infertile mouse testis, and the red fluorescence exhibited by the culture cell membranes was stably and continuously transmitted to the offspring. Next, via advanced mass spectrometry and an optimized proteomics platform, we constructed the proteome profile, with 682 proteins expressed in SSC s. Furthermore bioinformatics analysis showed that the list contained several known molecules that are regulated in SSC s. Several nucleoproteins and membrane proteins were chosen for further exploration using immunofluorescence and RT ‐ PCR . The results showed that SALL 1, EZH 2, and RCOR 2 are possibly involved in the self‐renewal mechanism of SSC s. Furthermore, the results of tissue‐specific expression analysis showed that Gpat2 and Pld6 were uniquely and highly expressed in mouse testes and cultured SSC s. The cellular localization of PLD 6 was further explored and the results showed it was primarily expressed in the spermatogonial membrane of mouse testes and cultured SSC s. The proteins identified in this study form the basis for further exploring the molecular mechanism of self‐renewal in SSC s and for identifying specific surface markers of SSC s.