Vimentin isoform expression in the human retina characterized with the monoclonal antibody 3CB2
Vimentin isoform expression in the human retina characterized with the monoclonal antibody 3CB2
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DOI：
10.1002/jnr.21623
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发表时间：
2008-06-01
影响因子：
4.2
通讯作者：
Prada, C.
中科院分区：
文献类型：
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作者：
Perez-Alvarez, M. J.;Isiegas, C.;Prada, C.
由单克隆抗体3CB 2（3CB 2-Ag和3CB 2 mAb）识别的抗原由脊椎动物的发育和成年脊椎动物中枢神经系统（CNS）中的放射状神经胶质细胞和星形胶质细胞以及神经干细胞表达。在这里，我们确定了3CB 2-Ag波形蛋白的蛋白质组学分析的人神经胶质细胞系U-87提取物（来自恶性星形细胞瘤）。事实上，3CB 2 mAb识别神经胶质细胞系中的三种波形蛋白亚型。在人视网膜中，3CB 2-Ag表达在Muller细胞，星形胶质细胞，一些血管，和水平细胞层中的细胞，通过免疫沉淀和免疫荧光法测定。免疫组织化学双标法可区分三种星形胶质细胞：波形蛋白（+）/GFAP（+）、波形蛋白（-）/GFAP（+）和波形蛋白（+）/GFAP（-）。因此，我们得出结论：1）3CB 2-Ag是波形蛋白; 2）波形蛋白亚型在正常和转化的星形胶质细胞中差异表达; 3）人视网膜星形胶质细胞显示分子异质性; 4）3CB 2 mAb是研究波形蛋白表达及其在人视网膜中功能的有价值工具。(c)2008 Wiley-Liss，Inc.
The antigen recognized by the monoclonal antibody 3CB2 (3CB2-Ag and 3CB2 mAb) is expressed by radial glia and astrocytes in the developing and adult vertebrate central nervous system (CNS) of vertebrates as well as in neural stem cells. Here we identified the 3CB2-Ag as vimentin by proteomic analysis of human glial cell line U-87 extracts (derived from a malignant astrocytoma). Indeed, the 3CB2 mAb recognized three vimentin isoforms in glial cell lines. In the human retina, 3CB2-Ag was expressed in Muller cells, astrocytes, some blood vessels, and cells in the horizontal cell layer, as determined by immunoprecipitation and immunofluorescence. Three populations of astrocytes were distinguishable by double-labeling immunohistochemistry: vimentin(+)/GFAP(+) vimentin(-)/GFAP(+) and vimentin(+)/GFAP(-). Hence, we conclude that 1) the 3CB2-Ag is vimentin; 2) vimentin isoforms are differentially expressed in normal and transformed astrocytes; 3) human retinal astrocytes display molecular heterogeneity; and 4) the 3CB2 mAb is a valuable tool to study vimentin expression and its function in the human retina. (c) 2008 Wiley-Liss, Inc.