Hansen G H, Hösli E, Belhage B, Schousboe A, Hösli L
Dept. of Biochemistry A, Panum Institute, Univ. of Copenhagen, Denmark.
Neurochem Res. 1991 Mar;16(3):341-6. doi: 10.1007/BF00966097.
GABAA-receptors were localized in explant cultures of rat cerebellum and in dissociated primary cultures of rat cerebellar granule cells and rat cerebellar astrocytes using the monoclonal antibody bd-17 directed against the beta-subunit of the GABAA/benzodiazepine/chloride channel complex. At the light microscope level specific staining of GABAA-receptors was localized in various types of neurones in explant cultures of rat cerebellum using the indirect peroxidase-antiperoxidase (PAP) technique, whereas no specific staining was found in astrocytes. At the electron microscope level labeling of GABAA-receptors was observed in the plasma membrane of both the cell bodies and processes in dissociated primary cultures of cerebellar granule cells using an indirect preembedding immunogold staining technique which in contrast to the classical PAP technique allows quantitative estimations to be performed. Quantification of the labeling intensity revealed a higher concentration of GABAA-receptors per microns plasma membrane in the cell bodies than in the processes. In discrete areas an extremely high density of the GABAA-receptors was observed. No specific labeling of GABAA-receptors was observed in dissociated primary cultures of cerebellar astrocytes.
使用针对GABAA/苯二氮䓬/氯化物通道复合物β亚基的单克隆抗体bd - 17，将GABAA受体定位在大鼠小脑的外植体培养物以及大鼠小脑颗粒细胞和大鼠小脑星形胶质细胞的解离原代培养物中。在光学显微镜水平，使用间接过氧化物酶 - 抗过氧化物酶（PAP）技术，GABAA受体的特异性染色定位于大鼠小脑外植体培养物中的各种神经元类型，而在星形胶质细胞中未发现特异性染色。在电子显微镜水平，使用间接包埋前免疫金染色技术，在小脑颗粒细胞解离原代培养物的细胞体和突起的质膜中观察到GABAA受体的标记，与经典的PAP技术相比，该技术允许进行定量估计。标记强度的定量显示，每微米质膜中细胞体的GABAA受体浓度高于突起中的浓度。在离散区域观察到GABAA受体的极高密度。在小脑星形胶质细胞的解离原代培养物中未观察到GABAA受体的特异性标记。
