Establishment of a proteome profile and identification of molecular markers for mouse spermatogonial stem cells.
Establishment of a proteome profile and identification of molecular markers for mouse spermatogonial stem cells.
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DOI：
10.1111/jcmm.12407
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发表时间：
2015-03
影响因子：
5.3
通讯作者：
Huang X
中科院分区：
文献类型：
--
作者：
Zhou Q;Guo Y;Zheng B;Shao B;Jiang M;Wang G;Zhou T;Wang L;Zhou Z;Guo X;Huang X
精原干细胞（SSCs）是一种未分化的细胞，在哺乳动物的整个生殖过程中维持精子发生所需。虽然SSC移植和培养提供了一个强有力的工具，以确定机制调节SSC功能，精确的信号转导机制管理SSC自我更新和特定的表面标记物纯化SSC仍有待明确确定。在本研究中，我们根据Shinohara实验室描述的方法建立了稳定的SSC培养。将培养的精原干细胞移植到不育小鼠睾丸中，可产生可育后代，且培养细胞膜上的红色荧光可稳定、持续地传递给后代。接下来，通过先进的质谱和优化的蛋白质组学平台，我们构建了蛋白质组图谱，其中682个蛋白质在SSC中表达。此外，生物信息学分析表明，该列表包含几个已知的在精原干细胞中调控的分子。几个核蛋白和膜蛋白的选择，进一步探索使用免疫荧光和RT-PCR。结果表明，SALL 1、EZH 2和RCOR 2可能参与了精原干细胞的自我更新机制。组织特异性表达分析结果表明，Gpat 2和Pld 6在小鼠睾丸和体外培养的精原干细胞中特异性高表达。进一步研究了PLD 6的细胞定位，结果显示PLD 6主要表达于小鼠睾丸精原细胞膜和培养的精原干细胞中。本研究中鉴定的蛋白质为进一步探索精原干细胞自我更新的分子机制和鉴定精原干细胞特异性表面标志物奠定了基础。
Spermatogonial stem cells (SSCs) 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 SSCs 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 SSCs 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 SSCs. Furthermore bioinformatics analysis showed that the list contained several known molecules that are regulated in SSCs. Several nucleoproteins and membrane proteins were chosen for further exploration using immunofluorescence and RT-PCR. The results showed that SALL1, EZH2, and RCOR2 are possibly involved in the self-renewal mechanism of SSCs. Furthermore, the results of tissue-specific expression analysis showed that Gpat2 and Pld6 were uniquely and highly expressed in mouse testes and cultured SSCs. The cellular localization of PLD6 was further explored and the results showed it was primarily expressed in the spermatogonial membrane of mouse testes and cultured SSCs. The proteins identified in this study form the basis for further exploring the molecular mechanism of self-renewal in SSCs and for identifying specific surface markers of SSCs.
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影响因子：
7.3
作者：
Fedele, M.;Franco, R.;Chieffi, P.
通讯作者：
Chieffi, P.
影响因子：
64.8
作者：
Guan, K;Nayernia, K;Hasenfuss, G
通讯作者：
Hasenfuss, G
影响因子：
30.8
作者：
Costoya, JA;Hobbs, RM;Pandolfi, PP
通讯作者：
Pandolfi, PP
影响因子：
64.5
作者：
MATSUI, Y;ZSEBO, K;HOGAN, BLM
通讯作者：
HOGAN, BLM
影响因子：
2.7
作者：
Eildermann, K.;Aeckerle, N.;Behr, R.
通讯作者：
Behr, R.