Black J A, Waxman S G, Friedman B, Elmer L W, Angelides K J
Department of Neurology, Yale University School of Medicine, New Haven, Connecticut 06510.
Glia. 1989;2(5):353-69. doi: 10.1002/glia.440020508.
Immuno-electron microscopic localization of sodium channels within astrocyte somata and processes of adult rat optic nerve was demonstrated with polyclonal antibody 7493. In immunoblots of crude glycoproteins from adult rat optic nerve, antisera 7493, which is directed against purified rat brain sodium channels, recognizes a 260 kDa protein. Antisera 7493 intensely immunostains axon membrane at nodes of Ranvier. Associated perinodal astrocyte processes are also stained with antisera 7493. In addition, astrocyte cell bodies and major processes exhibit immunoreactivity with antibody 7493. Immunostaining with antisera 7493 is heterogeneously distributed within astrocyte cytoplasm and also appears to be associated with some regions of astrocyte plasmalemma. Glial filaments are not immunostained with 7493 antisera. Astrocyte processes forming the glial limitans and surrounding blood vessels display reduced immunoreactivity to 7493 compared to longitudinally oriented or perinodal astrocyte processes. However, some focal regions of the glial limitans exhibit robust 7493 immunostaining. Oligodendrocytes do not display 7493 antisera immunoreactivity. Optic nerve sections incubated with preimmune sera or with 7493 antisera that had been previously adsorbed with purified sodium channel protein, exhibited no immunoreactivity. These results demonstrate localization of sodium channels within astrocytes in situ of rat optic nerve and extend previous electrophysiological and pharmacological findings of sodium channels in cultured astrocytes. Possible functional roles of sodium channels within astrocytes are discussed.
用多克隆抗体7493证明了成年大鼠视神经星形胶质细胞胞体和突起中钠通道的免疫电子显微镜定位。在成年大鼠视神经粗糖蛋白的免疫印迹中,针对纯化的大鼠脑钠通道的抗血清7493识别出一种260 kDa的蛋白质。抗血清7493强烈免疫染色郎飞结处的轴突膜。相关的结周星形胶质细胞突起也被抗血清7493染色。此外,星形胶质细胞胞体和主要突起对抗体7493表现出免疫反应性。抗血清7493的免疫染色在星形胶质细胞胞质内分布不均,似乎也与星形胶质细胞质膜的某些区域有关。胶质丝未被7493抗血清免疫染色。与纵向排列或结周的星形胶质细胞突起相比,形成胶质界膜和围绕血管的星形胶质细胞突起对7493的免疫反应性降低。然而,胶质界膜的一些局部区域表现出强烈的7493免疫染色。少突胶质细胞未显示7493抗血清免疫反应性。用免疫前血清或用先前已被纯化的钠通道蛋白吸附的7493抗血清孵育的视神经切片未表现出免疫反应性。这些结果证明了大鼠视神经原位星形胶质细胞中钠通道的定位,并扩展了先前关于培养星形胶质细胞中钠通道的电生理和药理学研究结果。文中讨论了星形胶质细胞中钠通道可能的功能作用。