MicroRNA-7a regulates Muller glia differentiation by attenuating Notch3 expression
MicroRNA-7a regulates Muller glia differentiation by attenuating Notch3 expression
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DOI：
10.1016/j.exer.2015.06.022
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发表时间：
2015-09-01
影响因子：
3.4
通讯作者：
Watanabe, Sumiko
中科院分区：
文献类型：
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作者：
Baba, Yukihiro;Aihara, Yuko;Watanabe, Sumiko
miRNA-7a 在健康和疾病的各个生物学方面发挥着关键作用。我们的目的是通过 miR-7a 的功能丧失和获得功能分析来揭示 miR-7a 在小鼠视网膜发育中的作用。将编码 miR-7a 或 miR-7a-decoy（反义 miR-7a）的质粒在 PO 时引入小鼠视网膜，并将视网膜作为外植体进行培养。然后，通过免疫染色检查视网膜祖细胞的增殖和视网膜亚型的分化。 miR-7a对视网膜祖细胞的增殖没有明显影响。然而，米勒胶质细胞标记物细胞周期蛋白D3的表达因miR-7a过表达而降低，并因miR-7a诱饵而上调，表明miR-7a负向调节米勒胶质细胞的分化。通过公共程序miRNA.org预测miR-7a的靶点，Notch3被认为是miR-7a靶点的候选基因之一。 Notch3 3' UTR 似乎包含与 miR-7a 种子序列互补的序列。使用含有 3'Notch UTR 潜在靶序列多次重复的质粒在 NIH3T3 细胞中进行报告基因检测，结果表明 miR-7a 通过 3'UTR 区域抑制报告基因 EGFP 的表达。视网膜中 sh-Notch3 的表达和 NICD3 的过表达表明 miR-7a 通过减弱 Notch3 的表达来调节 Muller 胶质细胞的分化。综上所述，我们发现 miR-7a 通过抑制 Notch3 表达来调节米勒胶质细胞的分化。 (C) 2015 Elsevier Ltd. 保留所有权利。
miRNA-7a plays critical roles in various biological aspects in health and disease. We aimed to reveal roles of miR-7a in mouse retinal development by loss- and gain-of-function analyses of miR-7a. Plasmids encoding miR-7a or miR-7a-decoy (anti-sense miR-7a) were introduced into mouse retina at PO, and the retina was cultured as explant. Then, proliferation of retinal progenitors and differentiation of retinal subtypes were examined by immunostaining. miR-7a had no apparent effect on the proliferation of retinal progenitor cells. However, the expression of Miller glia marker, cyclin D3, was reduced by miR-7a overexpression and up-regulated by miR-7a decoy, suggesting that miR-7a negatively regulates differentiation of Muller glia. Targets of miR-7a, which were predicted by using a public program miRNA.org, and Notch3 was suggested to be one of candidate genes of miR-7a target. Notch3 3' UTR appeared to contain complementary sequence to the seed sequence of miR-7a. A reporter assay in NIH3T3 cells using a plasmid containing multiple repeats of potential target sequence of 3' Notch UTR showed that miR-7a suppress expression of reporter EGFP through 3'UTR region. Expression of sh-Notch3 and overexpression of NICD3 in retina suggested that miR-7a regulates Muller glia differentiation through attenuation of Notch3 expression. Taken together, we revealed that the miR-7a regulates the differentiation of Miller glia through the suppression of Notch3 expression. (C) 2015 Elsevier Ltd. All rights reserved.