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Otx2同源框基因在源自成年虹膜和睫状体组织的细胞中诱导光感受器特异性表型。

Otx2 homeobox gene induces photoreceptor-specific phenotypes in cells derived from adult iris and ciliary tissue.

作者信息

Akagi Tadamichi, Mandai Michiko, Ooto Sotaro, Hirami Yasuhiko, Osakada Fumitaka, Kageyama Ryoichiro, Yoshimura Nagahisa, Takahashi Masayo

机构信息

Department of Ophthalmology and Visual Sciences, Graduate School of Medicine, Kyoto University, Kyoto, Japan.

出版信息

Invest Ophthalmol Vis Sci. 2004 Dec;45(12):4570-5. doi: 10.1167/iovs.04-0697.

Abstract

PURPOSE

It remains unclear which gene induction effectively generates photoreceptor-specific phenotypes from nonretinal tissues. The purpose of this study was to determine whether Crx and Otx2--homeobox genes related to photoreceptor development--can induce the generation of these phenotypes in cells derived from adult ciliary and iris tissue and in mesencephalon-derived neural stem cells.

METHODS

Crx and Otx2 were transferred into adult rat ciliary- and embryonic mesencephalon-derived neurospheres and adult rat iris-derived cells with the aid of a recombinant retrovirus. The presence of photoreceptor-specific phenotypes was confirmed by immunocytochemistry and Western blot analysis.

RESULTS

More than 90% of the Crx- and Otx2-transfected ciliary- and iris-derived cells exhibited rod opsin immunoreactivity, whereas few of the similarly transfected mesencephalon-derived neural stem cells expressed rod opsin. At least two additional key components of the phototransduction cascade, recoverin and Gdeltat1, were expressed by Crx- and Otx2-transfected iris-derived cells.

CONCLUSIONS

Crx and Otx2 effectively induced the generation of photoreceptor-specific phenotypes from ciliary- and iris-derived cells. That both Crx and Otx2 induced phenotype generation in cells derived from iris or ciliary tissue may suggest an approach to photoreceptor cell preparation for retinal transplantation.

摘要

目的

目前尚不清楚哪种基因诱导能有效地从非视网膜组织产生光感受器特异性表型。本研究的目的是确定与光感受器发育相关的同源框基因Crx和Otx2是否能在成年睫状体和虹膜组织来源的细胞以及中脑来源的神经干细胞中诱导这些表型的产生。

方法

借助重组逆转录病毒将Crx和Otx2导入成年大鼠睫状体和胚胎中脑来源的神经球以及成年大鼠虹膜来源的细胞。通过免疫细胞化学和蛋白质印迹分析确认光感受器特异性表型的存在。

结果

超过90%的转染Crx和Otx2的睫状体和虹膜来源细胞表现出视杆视蛋白免疫反应性,而同样转染的中脑来源神经干细胞中很少有细胞表达视杆视蛋白。转染Crx和Otx2的虹膜来源细胞表达了光转导级联的至少另外两个关键成分,恢复蛋白和Gδt1。

结论

Crx和Otx2有效地诱导了睫状体和虹膜来源细胞产生光感受器特异性表型。Crx和Otx2都能在虹膜或睫状体组织来源的细胞中诱导表型产生,这可能提示了一种用于视网膜移植的光感受器细胞制备方法。

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