A STUDY OF PROTEINS WITHIN THE CNS BY TWO-DIMENSIONAL GEL ELECTROPHORESIS

中枢神经系统中感兴趣的蛋白质的鉴定 双向凝胶电泳法(2DE)继续。1)我们有 一种命名为“蛋白质38”的可溶性蛋白质的分离纯化 在大鼠大脑中发现的蛋白质斑点之一 出现在2DE凝胶上。蛋白质38的分子量为 57kD(二聚体)和28.6kD(单体)。免疫细胞化学研究 使用针对该蛋白质的抗体显示，蛋白质38 定位于脑内的毛细血管内皮细胞， 外围设备。新生大鼠皮质来源的细胞培养 内皮内应激纤维染色呈阳性 细胞。2)第二项研究涉及隔离，部分 94kD蛋白的纯化及免疫组织化学定位 磷蛋白(“蛋白94”)。这种胞浆蛋白已经被 部分从牛尾状核中提纯。提纯 通过32P-磷酸化样品的放射自显影进行监测 用十二烷基硫酸钠-PAGE分离。使用针对该蛋白的兔抗体。 用于研究该蛋白的免疫组织化学定位 在新生大鼠皮质星形胶质细胞的细胞培养中发现 一种细胞类型，已初步确定为 小胶质细胞。3)对听觉核团内蛋白质的研究表明 一种蛋白质(相对分子质量=29kD，等电点=5.3)高度定位于 兔、大鼠和豚鼠的耳蜗核。

The identification of interesting proteins within the CNS utilizing 2-dimensional gel electrophoresis (2DE) continues. 1) We have isolated and purified a soluble protein designated "protein 38" which is found in the rat brain as one of the protein spots appearing on the 2DE gels. Protein 38 has a molecular weight of 57 kD (dimer) and 28.6 kD (monomer). Immunocytochemical studies using an antibody raised to this protein revealed that protein 38 is localizated in capillary endothelial cells in the brain and periphery. Cell cultures derived from newborn rat cortices revealed positive staining of stress fibers within endothelial cells. 2) A second study involves the isolation, partial purification and immunohistochemical localization of a 94 kD phosphoprotein ("protein 94"). This cytosolic protein has been partially purified from the bovine caudate nucleus. Purification was monitored by autoradiography of 32P-phosphorylated samples separated by SDS-PAGE. A rabbit antibody to this protein was used for a study of the immunohistochemical localization of this protein in cell cultures of astroglia of newborn rat cortices and revealed a single type of cell which has been tentatively identified as microglia. 3) Studies of proteins within auditory nuclei indicated one protein (MW = 29 kD, pI = 5.3) which is highly localized in the cochlear nuclei in rabbits, rats and guinea pigs.