DEVELOPMENTALLY-REGULATED AND SPATIALLY RESTRICTED ANTIGENS OF RADIAL GLIAL-CELLS
DEVELOPMENTALLY-REGULATED AND SPATIALLY RESTRICTED ANTIGENS OF RADIAL GLIAL-CELLS
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DOI：
10.1002/aja.1001970408
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发表时间：
1993-08-01
影响因子：
2.5
通讯作者：
SANES, JR
中科院分区：
文献类型：
--
作者：
HERMAN, JP;VICTOR, JC;SANES, JR
放射状胶质细胞存在于胚胎脊椎动物中枢神经系统（CNS）的许多部位，参与引导成神经细胞从脑室区到达其层流目的地。此外，放射状胶质细胞可能是某些CNS神经元和胶质细胞的祖细胞。为了获得新的见解，这些细胞的结构和发展，我们已经产生和特点的单克隆抗体，识别鸡视顶盖放射状胶质细胞的面板。用胚胎第10天（E）顶盖的匀浆免疫小鼠，并通过免疫荧光和免疫印迹分析抗体。我们在这里描述三对抗体。1)H5和先前产生的抗体R5（Drager等人，J. Neurosci. 4：2025，1984），从E7至E20染色放射状神经胶质细胞的整个范围。在E10 tecta制备的培养物中，两者都染色胶质细胞中的丝状网状结构，但不染色神经元中的丝状网状结构。在免疫印迹中，两者都识别与波形蛋白密切相关（或相同）的类似于52 kD的蛋白质。2)H28和H29在E7和E14之间染色放射状胶质细胞，但不晚于E7和E14。此外，H28和H29染色在脑室和中间区比在顶盖板的层中明显更强。这两种抗体都识别培养的神经胶质细胞中的细胞内表位和免疫印迹上类似于35 kD的蛋白质。3)H2和H27识别集中在晚期（E16-E20）胚胎中放射状胶质细胞最表面突起和尾足中的抗原。它们在培养的神经胶质细胞中染色不同的结构，表明它们识别不同的抗原。H27在免疫印迹上识别类似于29 kD的蛋白质。因此，抗体H5和R5是放射状神经胶质细胞在所有阶段的良好标志物，而其他定义的抗原是发育调节和定位于离散域。总之，这些抗体可用于研究放射状胶质细胞的时间和空间特化。
Radial glial cells, present in many parts of the embryonic vertebrate central nervous system (CNS), have been implicated in the guidance of neuroblasts from the ventricular zone to their laminar destinations. Moreover, radial glial cells may be progenitors of some CNS neurons and glia. To gain new insight into the structure and development of these cells, we have generated and characterized a panel of monoclonal antibodies that recognize radial glial cells of the chick optic tectum. Mice were immunized with homogenates of embryonic day (E) 10 tectum, and antibodies were analyzed by immunofluorescence and immunoblotting. We describe here three pairs of antibodies. 1) H5 and a previously generated antibody, R5 (Drager et al., J. Neurosci. 4:2025, 1984), stain the whole extent of the radial glial cell from E7 to E20. In cultures prepared from E10 tecta, both stain a filamentous meshwork in glial cells but not in neurons. On immunoblots, both recognize a protein of similar to 52 kD that is closely related (or identical) to vimentin. 2) H28 and H29 stain radial glia between E7 and E14, but not later. Moreover, H28 and H29 staining is markedly more intense in the ventricular and intermediate zones than in the laminae of the tectal plate. Both of these antibodies recognize an intracellular epitope in cultured glial cells and a protein of similar to 35 kD on immunoblots. 3) H2 and H27 recognize antigens concentrated in the most superficial processes and endfeet of radial glia in late (E16-E20) embryos. They stain distinct structures in cultured glia, suggesting that they recognize distinct antigens. H27 recognizes a protein of similar to 29 kD on immunoblots. Thus antibodies H5 and R5 are good markers of radial glial cells at all stages, whereas the others define antigens that are developmentally regulated and localized to discrete domains. Together, these antibodies can be used to study temporal and spatial specializations of radial glia.