Dicer Plays Essential Roles for Retinal Development by Regulation of Survival and Differentiation
Dicer Plays Essential Roles for Retinal Development by Regulation of Survival and Differentiation
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DOI：
10.1167/iovs.10-6428
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发表时间：
2011-05-01
影响因子：
4.4
通讯作者：
Watanabe, Sumiko
中科院分区：
文献类型：
--
作者：
Iida, Atsumi;Shinoe, Toru;Watanabe, Sumiko
目的。MicroRNA在发育和生物学过程中的作用已经引起了人们的广泛关注。在各种器官的细胞生存和增殖中，DICER起着至关重要的作用。我们通过对小鼠视网膜特异的条件基因敲除来检测DICER在视网膜发育中的作用。方法：Dkk3-Cre在胚胎早期的视网膜祖细胞中表达Cre基因。作者分析了DKK-CRE/DICER-FLOX(DICER-CKO)小鼠的存活、增殖和分化情况。通过电穿孔的方法将Cre表达载体导入新生小鼠视网膜，观察其视网膜分化情况。结果：基于孟德尔遗传学，Dird-CKO小鼠按我们预期的数量出生，但它们的眼睛一直没有睁开。视网膜前体细胞在胚胎发育过程中大量死亡，导致小眼球，出生后14天大部分视网膜细胞消失。体外重组培养的DICER-CKO视网膜细胞显示，细胞死亡和DICER失活抑制增殖是以细胞自主的方式发生的。细胞分化标记物在视网膜中有表达，但这些细胞定位异常，内网状层缺失，提示细胞迁移和形态分化，尤其是突起伸展受到干扰。强迫新生儿表达Cre诱导细胞凋亡并影响分化标记物的表达。结论综上所述，这些结果表明在视网膜早期发育过程中，Dird是必不可少的。(投资眼科VS科学。2011年；52：3008-3017)doi：10.1167/iovs.10-6428
PURPOSE. Much attention has been paid to the roles of microRNA in developmental and biological processes. Dicer plays essential roles in cell survival and proliferation in various organs. We examined the role of Dicer in retinal development using retina-specific conditional knockout of Dicer in mice.METHODS. Dkk3-Cre expressed the Cre gene in retinal progenitor cells from an early embryonic stage. The authors analyzed Dkk-Cre/Dicer-flox (Dicer-CKO) mice for their survival, proliferation, and differentiation. To analyze the role of Dicer in later stages of retinal development, a Cre expression plasmid was introduced into the neonatal retina by electroporation, and retinal differentiation was examined.RESULTS. Dicer-CKO mice were born at the numbers we expected, based on Mendelian genetics, but their eyes never opened. Massive death of retinal progenitor cells occurred during embryogenesis, resulting in microphthalmia, and most retinal cells had disappeared by postnatal day 14. In vitro reaggregation culture of Dicer-CKO retinal cells showed that cell death and the suppression of proliferation by Dicer inactivation occurred in a cell-autonomous manner. Cell differentiation markers were expressed in the Dicer-CKO retina; however, these cells localized abnormally, and the inner plexiform layer was absent, suggesting that cell migration and morphologic differentiation, especially process extension, were perturbed. Forced neonatal expression of Cre induced apoptosis and affected the expression of differentiation markers.CONCLUSIONS. Taken together, these results show that Dicer is essential during early retinal development. (Invest Ophthalmol Vis Sci. 2011;52:3008-3017) DOI:10.1167/iovs.10-6428