Unraveling the proteomic profile of mice testis during the initiation of meiosis.
Unraveling the proteomic profile of mice testis during the initiation of meiosis.
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揭示减数分裂开始期间小鼠睾丸的蛋白质组谱。
DOI：
10.1016/j.jprot.2015.02.015
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发表时间：
2015-04
期刊：
影响因子：
--
通讯作者：
sha j
中科院分区：
文献类型：
--
作者：
zhou t;guo x;huang x;sha j
在小鼠中，一旦原始生殖细胞（PGCs）产生，它们就会继续增殖并迁移，最终到达未来的性腺。它们在生殖腺定植后开始性分化。在此过程中，视黄酸（RA）诱导减数分裂的雌性生殖细胞，这进行到双线期阶段的减数分裂前期I，而男性生殖细胞启动生长停滞。出生后，小鼠精原细胞减数分裂开始，转化为细线期前精母细胞。有证据表明RA在精原细胞分化和减数分裂启动中起着重要的调节作用。然而，它仍然是不太清楚什么响应RA和RA信号如何参与减数分裂。因此，我们构建了小鼠睾丸发育过程中减数分裂起始相关蛋白的蛋白质组学图谱，并鉴定了104个差异表达蛋白（≥ 1.5倍）。生物信息学分析显示蛋白质在特定的细胞过程中发挥作用。通过Western blot分析，我们发现Tfr c基因在精原干细胞（SSC）中具有RA反应性，并含有RA反应元件，RA可上调Tfr c基因表达。生物学意义精子发生包括精原细胞的有丝分裂、精母细胞的减数分裂和精子发生，其中减数分裂是生殖细胞特有的事件，而不是体细胞特有的事件。到目前为止，有丝分裂向减数分裂转变的详细分子机制仍不清楚。利用高通量蛋白质组学技术，现在可以系统地鉴定可能涉及的蛋白质。利用基于TMT-6plex的定量，我们在没有减数分裂的睾丸（第8.5天）和减数分裂开始的睾丸（第10.5天）之间鉴定了104种差异蛋白。在本研究中还发现了一种成熟分裂起始所必需的蛋白stra 8的差异表达。生物信息学分析和功能研究揭示了几种受视黄酸调控的蛋白质，视黄酸是一种已知的调节减数分裂起始的化学物质。因此，这种定量蛋白质组学的方法可以确定减数分裂起始调控蛋白，这些蛋白质的进一步功能研究将有助于阐明减数分裂起始的机制。
In mice, once primordial germ cells (PGCs) are generated, they continue to proliferate and migrate to eventually reach the future gonads. They initiate sexual differentiation after their colonization of the gonads. During this process, retinoic acid (RA) induces meiosis in the female germ cells, which proceeds to the diplotene stage of meiotic prophase I, whereas the male germ cells initiate growth arrest. After birth, meiosis is initiated in mice spermatogonia by their conversion to preleptotene spermatocytes. There are evidences showing the roles of RA in the regulation of spermatogonial differentiation and meiosis initiation. However, it is still not well known on what responds to RA and how RA signaling engages meiosis. Thus, we constructed a proteomic profile of proteins associated with meiosis onset during testis development in mouse and identified 104 differentially expressed proteins (≥ 1.5 folds). Bioinformatic analysis showed proteins functioning in specific cell processes. The expression patterns of five selected proteins were verified via Western blot, of which we found thatTfrcgene was RA responsive, with a RA responsive element, and could be up regulated by RA in spermatogonial stem cell (SSC) line. Taken together, the results provide an important reference profile for further functional study of meiosis initiation.Biological significanceSpermatogenesis involves mitosis of spermatogonia, meiosis of spermatocytes and spermiogenesis, in which meiosis is a unique event to germ cells, and not in the somatic cells. Till now, the detailed molecular mechanisms of the transition from mitosis to meiosis are still not elucidated. With high-throughput proteomic technology, it is now possible to systemically identify proteins possibly involved. With TMT-6plex based quantification, we identified 104 proteins differentially between testes without meiosis (day 8.5) and those that were meiosis initiated (day 10.5). And a well-known protein essential for meiosis initiation, stra8, was identified to be differentially expressed in the study. And bioinformatic analysis and functional studies revealed several proteins regulated by retinoic acid, a chemical known to regulate the meiosis initiation. Thus, this quantitative proteomic approach can identify meiosis initiation regulating proteins, and further functional studies of these proteins will help elucidate the mechanisms of meiosis initiation.
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影响因子：
3.4
作者：
Deutsch, Eric W.;Mendoza, Luis;Shteynberg, David;Farrah, Terry;Lam, Henry;Tasman, Natalie;Sun, Zhi;Nilsson, Erik;Pratt, Brian;Prazen, Bryan;Eng, Jimmy K.;Martin, Daniel B.;Nesvizhskii, Alexey I.;Aebersold, Ruedi
通讯作者：
Aebersold, Ruedi
影响因子：
--
作者：
Cash AC;Andrews J
通讯作者：
Andrews J
影响因子：
3.6
作者：
de Rooij, DG;Okabe, M;Nishimune, Y
通讯作者：
Nishimune, Y
影响因子：
64.5
作者：
Duester G
通讯作者：
Duester G
影响因子：
56.9
作者：
Bowles, J;Knight, D;Koopman, P
通讯作者：
Koopman, P