Purification of -aminobutyric acid (GABA) receptor and formation of monoclonal antibody against the purified receptor.

采用亲和柱层析法，以1012-s苯二氮卓为固定配体，从牛大脑皮层中纯化了可溶性-氨基丁酸（GABA）和苯二氮卓（BZP）受体。用Sephadex G-200对纯化的GABA/BZP受体复合物进行凝胶过滤，其分子量约为340,000。1012- s洗脱组分的sds -聚丙烯酰胺凝胶电泳图谱显示存在两条分子量约为53,000和57,000的主要条带，这两条条带都被[^3H]氟硝西泮不可逆地光亲和标记。纯化后的gaba受体为一种糖蛋白，其杂化型碳水化合物链参与gaba受体与BZP受体的功能偶联。双周皮下注射纯化的GABA受体对白化病家兔形成的GABA受体抗体免疫组织化学表明，该受体复合物在大鼠脑内分布于不同的脑区，如小脑浦肯野细胞和颗粒细胞层、黑质致密部大细胞、大脑皮层（III-V层）锥体细胞和海马。

Purification of solubilized -aminobutyric acid (GABA) and benzodiazepine (BZP) receptors from the bovine cerebral cortex was exployes by the affinity columu chromatography using a benzo-diazepine, 1012-s, as immobilized ligand. The gel filtration using Sephadex G-200 of the pufified GABA/BZP receptor complex revealed the molecular weight of the receptor complex was approximately 340,000. SDS-polyacrylamide gel electrophoretic profiles of the 1012-S-eluted fraction showed the existence of two major bands having the molecular weights of approximately 53,000 and 57,000, both of which were irreversibly photoaffinity-labeled with [^3H]flunitrazepam. The purified gaba receptor was found to be a glycoprotein and its carbohydrate chain of a hybrid type was involved in the functional coupling between GABA receptor and BZP receptor. The GABA receptor antibody formed by biweekly subcutaneous injection of the purified GABA receptor to Albino rabbits has revealed immunohistochemically that this receptor complex in the rat brain is localized various brain areas such as the somata of Purkinje cells and gramule cells layer of the cerebellum, large cells in the pars compacta of substantia nigra, pyramidal cells in the cerebral cortex (layeers III-V) and hippocampus, respectively.