Iris-derived cells from adult rodents and primates adopt photoreceptor-specific phenotypes
Iris-derived cells from adult rodents and primates adopt photoreceptor-specific phenotypes
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DOI：
10.1167/iovs.04-1112
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发表时间：
2005-09-01
影响因子：
4.4
通讯作者：
Takahashi, M
中科院分区：
文献类型：
--
作者：
Akagi, T;Akita, J;Takahashi, M
目的.本研究的目的是探讨视网膜感光细胞发育相关基因对啮齿类和灵长类动物虹膜细胞的影响，以及虹膜细胞作为视网膜移植供体细胞的可能性。成年大鼠和猴虹膜组织培养在无血清培养基中含有碱性成纤维细胞生长因子。用重组逆转录病毒进行Crx、Nrl、NeuroD和一些组合（Crx-Nrl、Crx-NeuroD）的基因递送。采用免疫细胞化学、Western blot、RT-PCR和细胞内记录等方法检测转基因后虹膜细胞中光感受器特异性表型的表达。将虹膜衍生细胞与胚胎视网膜外植体共培养，以研究这些细胞在共培养条件下的潜在整合。Crx的错误表达诱导成年大鼠虹膜细胞表达几种光感受器特异性抗原和转录物，如视紫红质、恢复素、cGMP门控通道、arrestin、光感受器间视网膜结合蛋白、视紫红质激酶和NeuroD。在灵长类动物中，需要Crx和NeuroD的组合来诱导猴虹膜衍生的细胞采用光感受器特异性表型。此外，来自大鼠和灵长类动物虹膜组织的光感受器样细胞表现出视杆光感受器特异性的电生理反应，光刺激后Crx和Crx-NeuroD基因转移，分别。结果进一步表明，虹膜源性细胞在共培养条件下整合到发育中的宿主视网膜中。啮齿类和灵长类动物的成年虹膜衍生培养细胞通过转录因子的诱导表达光感受器特异性表型。这些虹膜衍生的光感受器样细胞具有视杆细胞光感受器的电生理特征。此外，它们可以在共培养条件下整合到发育中的视网膜中。
PURPOSE. The purpose of this study was to investigate the effects of various genes related to photoreceptor development on rodent and primate iris cells and the potential of iris cells as donor cells for retinal transplantation.METHODS. Adult rat and monkey iris tissue were cultured in serum-free medium containing basic fibroblast growth factor. Gene deliveries of Crx, Nrl, NeuroD and some combinations (Crx-Nrl, Crx-NeuroD) were performed with recombinant retrovirus. Immunocytochemistry, Western blot analysis, RT-PCR, and intracellular recording were used to examine the expression of photoreceptor-specific phenotypes in the iris-derived cells after gene transfer,. Coculture of the iris-derived cells with embryonic retinal explant was conducted, to investigate the potential integration of these cells in coculture conditions.RESULTS. Misexpression of Crx induced adult rat iris cells to express several photoreceptor-specific antigens and transcripts, such as rhodopsin, recoverin, cGMP-gated channel, arrestin, interphotoreceptor retinal-binding protein, rhodopsin kinase, and NeuroD. In primates, a combination of Crx and NeuroD was needed to induce monkey iris - derived cells to adopt photoreceptor-specific phenotypes. Furthermore, the photoreceptor-like cells derived from both rat- and primate-iris tissues showed rod photoreceptor-specific electrophysiological response to light stimuli after Crx and Crx-NeuroD gene transfer, respectively. The results further showed that iris-derived cells integrated in the developing host retina in coculture conditions.CONCLUSIONS. Adult iris-derived cultured cells of both rodents and primates expressed photoreceptor-specific phenotypes by inductions of transcription factors. These iris-derived photoreceptor-like cells have electrophysiological characteristics of rod photoreceptors. Furthermore, they can integrate in the developing retina under coculture conditions.