Nodal signaling via an autocrine pathway promotes proliferation of mouse spermatogonial stem/progenitor cells through Smad2/3 and Oct-4 activation.
Nodal signaling via an autocrine pathway promotes proliferation of mouse spermatogonial stem/progenitor cells through Smad2/3 and Oct-4 activation.
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DOI：
10.1002/stem.198
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发表时间：
2009-10
期刊：
影响因子：
5.2
通讯作者：
Dym, Martin
中科院分区：
文献类型：
--
作者：
He, Zuping;Jiang, Jiji;Kokkinaki, Maria;Dym, Martin
精子发生是精原干细胞分裂和分化为精子的过程。然而，控制其命运的自分泌分子和信号通路仍然未知。本研究旨在鉴定调节精原干/祖细胞增殖、分化和存活的新型生长因子和信号通路。为此，我们首次探讨了TGFβ超家族成员Nodal在小鼠精原干/祖细胞中的表达、功能和信号通路。我们证明，Nodal及其受体都存在于这些细胞和精原干/祖细胞系（C18-4细胞），而Nodal是未检测到的支持细胞或分化的生殖细胞，通过RT-PCR，Western印迹和免疫细胞化学分析。Nodal促进精原干/祖细胞和C18-4细胞的增殖，而Nodal受体抑制剂SB 431542阻断其增殖，如增殖和溴脱氧尿苷掺入试验所示。如TUNEL和增殖测定所示，通过RNA干扰的结敲低导致细胞凋亡的显著增加和细胞分裂的减少。相反，Nodal的过表达导致细胞增殖的增加。Nodal激活Smad 2/3磷酸化、Oct-4转录、细胞周期蛋白D1和细胞周期蛋白E表达，而SB 431542完全消除它们的增加。总之，Nodal被鉴定为第一个通过Smad 2/3和Oct-4激活促进小鼠精原干/祖细胞增殖的自分泌信号分子。因此，这项研究提供了新的和重要的见解调节精原干/祖细胞的增殖和存活的分子机制。
Spermatogenesis is the process that involves the division and differentiation of spermatogonial stem cells into spermatozoa. However, the autocrine molecules and signaling pathways controlling their fate remain unknown. This study was designed to identify novel growth factors and signaling pathways that regulate proliferation, differentiation, and survival of spermatogonial stem/progenitor cells. To this end, we have for the first time explored the expression, function, and signaling pathway of Nodal, a member of the TGFβ superfamily, in mouse spermatogonial stem/progenitor cells. We demonstrate that both Nodal and its receptors are present in these cells and in a spermatogonial stem/progenitor cell line (C18-4 cells), whereas Nodal is undetected in Sertoli cells or differentiated germ cells, as assayed by RT-PCR, Western blots, and immunocytochemistry. Nodal promotes proliferation of spermatogonial stem/progenitor cells and C18-4 cells, while Nodal receptor inhibitor SB431542 blocks their propagation as shown by proliferation and bromodeoxyuridine incorporation assays. Nodal knockdown by RNA interference results in a marked increase of cell apoptosis and a reduction of cell division as indicated by TUNEL and proliferation assays. Conversely, overexpression of Nodal leads to an increase of cell proliferation. Nodal activates Smad2/3 phosphorylation, Oct-4 transcription, cyclin D1, and cyclin E expression, whereas SB431542 completely abolishes their increase. Together, Nodal was identified as the first autocrine signaling molecule that promotes proliferation of mouse spermatogonial stem/progenitor cells via Smad2/3 and Oct-4 activation. This study thus provides novel and important insights into molecular mechanisms regulating proliferation and survival of spermatogonial stem/progenitor cells.
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