Herman J P, Victor J C, Sanes J R
Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, Missouri 63110.
Dev Dyn. 1993 Aug;197(4):307-18. doi: 10.1002/aja.1001970408.
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 (Dräger 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 approximately 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 approximately 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 approximately 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.
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,与神经母细胞从脑室区到其层状目的地的引导有关。此外,放射状胶质细胞可能是一些中枢神经系统神经元和神经胶质细胞的祖细胞。为了深入了解这些细胞的结构和发育,我们制备并鉴定了一组识别鸡视顶盖放射状胶质细胞的单克隆抗体。用胚胎第10天(E10)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。我们在此描述三对抗体。1)H5和先前制备的抗体R5(Dräger等人,《神经科学杂志》4:2025,1984),从E7到E20对放射状胶质细胞的整个范围进行染色。在从E10顶盖制备的培养物中,两者都对胶质细胞中的丝状网络进行染色,但对神经元则不染色。在免疫印迹上,两者都识别一种与波形蛋白密切相关(或相同)的约52 kD的蛋白质。2)H28和H29在E7到E14之间对放射状胶质细胞进行染色,但之后则不染色。此外,H28和H29染色在脑室区和中间区明显比顶盖板层更强烈。这两种抗体都识别培养的胶质细胞中的细胞内表位以及免疫印迹上一种约35 kD的蛋白质。3)H2和H27识别晚期(E16 - E20)胚胎中集中在放射状胶质细胞最表面的突起和终足中的抗原。它们在培养的胶质细胞中染色不同的结构,表明它们识别不同的抗原。H27在免疫印迹上识别一种约29 kD的蛋白质。因此,抗体H5和R5是放射状胶质细胞在所有阶段的良好标记物,而其他抗体则定义了在发育过程中受到调节并定位于离散区域的抗原。这些抗体一起可用于研究放射状胶质细胞的时间和空间特化。