MicroRNA-202 maintains spermatogonial stem cells by inhibiting cell cycle regulators and RNA binding proteins.
MicroRNA-202 maintains spermatogonial stem cells by inhibiting cell cycle regulators and RNA binding proteins.
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MicroRNA-202 通过抑制细胞周期调节因子和 RNA 结合蛋白来维持精原干细胞
DOI：
10.1093/nar/gkw1287
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发表时间：
2017-04-20
影响因子：
14.9
通讯作者：
Han C
中科院分区：
文献类型：
--
作者：
Chen J;Cai T;Zheng C;Lin X;Wang G;Liao S;Wang X;Gan H;Zhang D;Hu X;Wang S;Li Z;Feng Y;Yang F;Han C
摘要miRNAs在哺乳动物精子发生过程中起重要作用。然而，大多数miRNAs在精子发生中的功能和潜在机制仍然未知。在这里，我们报告了miR-202在小鼠精原干细胞（SSCs）中的高表达，并且受到胶质细胞源性神经营养因子（GDNF）和视黄酸（RA）的反向调节，这两个因素是SSC自我更新和分化的关键因素。我们使用诱导型CRISPR-Cas9敲除培养的SSC中的miR-202，并发现敲除的SSC引发了过早分化，伴随着干细胞活性降低和有丝分裂和凋亡增加。通过基于iTRAQ的蛋白质组学分析和RNA测序鉴定靶基因，并且靶基因富含细胞周期调节因子和RNA结合蛋白。发现Rbfox 2和Cpeb 1是miR-202的直接靶点，Rbfox 2而不是Cpeb 1，对于SSCs分化为减数分裂细胞是必需的。因此，已经鉴定了由GDNF和RA的信号分子、miR-202和多种下游效应物组成的SSC命运调控网络。
Abstract miRNAs play important roles during mammalian spermatogenesis. However, the function of most miRNAs in spermatogenesis and the underlying mechanisms remain unknown. Here, we report that miR-202 is highly expressed in mouse spermatogonial stem cells (SSCs), and is oppositely regulated by Glial cell-Derived Neurotrophic Factor (GDNF) and retinoic acid (RA), two key factors for SSC self-renewal and differentiation. We used inducible CRISPR-Cas9 to knockout miR-202 in cultured SSCs, and found that the knockout SSCs initiated premature differentiation accompanied by reduced stem cell activity and increased mitosis and apoptosis. Target genes were identified with iTRAQ-based proteomic analysis and RNA sequencing, and are enriched with cell cycle regulators and RNA-binding proteins. Rbfox2 and Cpeb1 were found to be direct targets of miR-202 and Rbfox2 but not Cpeb1, is essential for the differentiation of SSCs into meiotic cells. Accordingly, an SSC fate-regulatory network composed of signaling molecules of GDNF and RA, miR-202 and diverse downstream effectors has been identified.