Sox2 Plays a Role in the Induction of Amacrine and Muller Glial Cells in Mouse Retinal Progenitor Cells
Sox2 Plays a Role in the Induction of Amacrine and Muller Glial Cells in Mouse Retinal Progenitor Cells
复制标题
DOI：
10.1167/iovs.07-1619
复制
发表时间：
2009-01-01
影响因子：
4.4
通讯作者：
Watanabe, Sumiko
中科院分区：
文献类型：
--
作者：
Lin, Ya-Ping;Ouchi, Yasuo;Watanabe, Sumiko
目的。转录因子Sox2在人和小鼠视网膜发育中都起着重要作用。虽然在小鼠中已经研究了Sox2的功能丧失突变，但在神经视网膜中缺乏功能获得的实验。用免疫组织化学方法检测Sox2在发育小鼠视网膜中的详细表达谱。然后，通过逆转录病毒介导的基因转移，在E17小鼠胚胎视网膜外植体培养物中表达Sox2，研究其在视网膜发育中的作用。此外，利用逆转录病毒介导的系统，shRNA在视网膜外植体中抑制Sox2。Sox2在胚胎视网膜的整个神经母细胞层表达，而在成熟视网膜中仅在内核层表达。双免疫染色显示Sox2在Muller胶质细胞和一部分无毛细胞中表达。在小鼠视网膜外植体培养中，强制表达Sox2导致内核层无毛细胞的大量积累；此外，在外核层的最内侧也发现了表达无分泌细胞标记的细胞。在表达Sox2的细胞中观察到Pax6的表达，Sox2激活Pax6启动子驱动Y79细胞中荧光素酶的表达。Pax6在无腺细胞分化中起重要作用。视网膜祖细胞增殖的减少伴随着这些影响。shRNA抑制Sox2表达可导致内核层细胞数量减少。因此，异位表达Sox2可以诱导小鼠视网膜从E17期开始无分泌细胞，可能是通过促进细胞周期退出。（中国眼科杂志，2009;50:68-74）DOI: 10.1167/iovs.07-1619
PURPOSE. The transcription factor Sox2 plays important roles in both human and mouse retinal development. Although loss-of-function mutations in Sox2 have been studied in mice, gain-of-function experiments in the neural retina have been lacking.METHODS. The detailed expression pattern of Sox2 in the developing mouse retina was examined by immunohistochemistry. Then, Sox2 was expressed in a retinal explant culture prepared from E17 mouse embryos by retrovirus-mediated gene transfer to examine its role in retinal development. In addition, shRNA was used to suppress Sox2 in a retinal explant using a retrovirus-mediated system.RESULTS. Sox2 was expressed throughout the neuroblastic layer in the embryonic retina, but only in the inner nuclear layer in the mature retina. Double immunostaining revealed that Sox2 was expressed in Muller glial cells and in a subset of amacrine cells. Forced expression of Sox2 in a mouse retinal explant culture resulted in the dramatic accumulation of amacrine cells in the inner nuclear layer; in addition, cells expressing amacrine cell markers were also found on the innermost side of the outer nuclear layer. The expression of Pax6, which plays an important role in amacrine cell differentiation, was observed in the Sox2-expressing cells, and Sox2 activated the Pax6 promoter to drive luciferase expression in Y79 cells. A decrease in retinal progenitor cell proliferation accompanied these effects. The suppression of Sox2 expression by shRNA resulted in a decreased number of cells in the inner nuclear layer.CONCLUSIONS. Therefore, ectopic Sox2 expression can induce amacrine cells in the mouse retina from stage E17 onward, possibly by facilitating cell cycle exit. (Invest Ophthalmol Vis Sci. 2009; 50: 68-74) DOI: 10.1167/iovs.07-1619