Expression of cone-photoreceptor-specific antigens in a cell line derived from retinal tumors in transgenic mice
Expression of cone-photoreceptor-specific antigens in a cell line derived from retinal tumors in transgenic mice
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DOI：
10.1167/iovs.03-1114
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发表时间：
2004-03-01
影响因子：
4.4
通讯作者：
Al-Ubaidi, MR
中科院分区：
文献类型：
--
作者：
Tan, E;Ding, XQ;Al-Ubaidi, MR
目的.目的：检测永生化小鼠视网膜细胞系（661 W）的感光细胞标志物，验证其感光细胞来源。661 W细胞克隆自转基因小鼠系的视网膜肿瘤，该转基因小鼠系在人光感受器间视黄醇结合蛋白（IRBP）启动子的控制下表达猿猴病毒（SV）40 T抗原。进行形态学、免疫细胞化学和免疫印迹分析以表征这些细胞。总细胞蛋白用于各种光感受器特异性蛋白的免疫印迹分析。661 W细胞呈单层生长，并显示出神经元细胞的特征性过程。免疫印迹分析显示661 W细胞表达SV 40 T抗原、蓝色和绿色锥状色素、转导素和锥状抑制素。蓝、绿色视蛋白的免疫细胞化学检测显示分布于整个细胞，包括细胞核。然而，这些细胞不表达视杆细胞特异性抗原，如视蛋白和抑制蛋白，或视杆细胞和视锥细胞特异性蛋白，如视球蛋白、外周蛋白/rds和ROM 1。此外，细胞不表达RPE 65，一种视锥细胞和RPE细胞特异性蛋白。661 W细胞表现出视锥光感受器细胞所表现出的细胞和生化特征。这些细胞也类似于神经元细胞与他们的纺锤形进程，并应作为一个有用的替代体外模型的视锥细胞生物学和相关疾病的研究。
PURPOSE. To examine an immortalized mouse retinal cell line (661W) for markers characteristic of photoreceptor cells and validate its photoreceptor origin.METHODS. The 661W cells were cloned from retinal tumors of a transgenic mouse line that expresses the simian virus (SV) 40 T antigen under control of the human interphotoreceptor retinol-binding protein (IRBP) promoter. Morphologic, immunocytochemical, and immunoblot analyses were performed to characterize these cells. Total cellular protein was used for immunoblot analysis of various photoreceptor-specific proteins.RESULTS. 661W cells grew as a monolayer and exhibited processes characteristic of neuronal cells. Immunoblot analysis showed that 661W cells expressed SV40 T antigen, blue and green cone pigments, transducin, and cone arrestin. Immunocytochemical detection of blue and green opsins showed distribution throughout the cell, the nucleus included. However, these cells did not express rod-specific antigens, such as opsin and arrestin or rod- and cone-specific proteins such as phosducin, peripherin/rds, and ROM1. Furthermore, the cells did not express RPE65, a cone- and RPE-cell-specific protein.CONCLUSIONS. 661W cells demonstrate cellular and biochemical characteristics exhibited by cone photoreceptor cells. These cells also resemble neuronal cells with their spindlelike processes and should serve as a useful alternative in vitro model for the study of cone photoreceptor cell biology and associated diseases.