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猫视觉皮层中星形胶质细胞GABAA受体的免疫细胞化学检测

Immunocytochemical detection of astrocyte GABAA receptors in cat visual cortex.

作者信息

Rosier A, Arckens L, Orban G A, Vandesande F

机构信息

Laboratory of Neuroendocrinology and Immunological Biotechnology, Katholieke Universiteit Leuven, Belgium.

出版信息

J Histochem Cytochem. 1993 May;41(5):685-92. doi: 10.1177/41.5.8385682.

Abstract

A nine amino-acid peptide derived from the beta 1-subunit of the bovine GABAA receptor was used for immunization of mice and subsequent production of monoclonal antibodies (MAb). In view of the later immunocytochemical application of the MAb to sections of cat visual cortex, the MAb were characterized on similar tissue. The GABAA receptor was isolated by affinity chromatography of protein material obtained from cat cortical gray matter. The antibodies were characterized by SDS-PAGE, followed by immunoblotting and immunoadsorption. Immunocytochemical staining with the MAb revealed labeled cells throughout all layers of the cat visual cortex as well as within the white matter. The morphology of most stained cells in the white matter and of some cells in the cortical gray matter closely resembled that of astrocytes. Double immunocytochemical staining with an antiserum against glial fibrillary acidic protein (GFAP), followed by light microscopic examination, indeed confirmed that astrocytes in both white and gray matter in the cat cortex possess GABAA receptors.

摘要

一种源自牛GABAA受体β1亚基的九氨基酸肽被用于免疫小鼠并随后制备单克隆抗体(MAb)。鉴于随后将MAb用于猫视觉皮层切片的免疫细胞化学应用,这些MAb在类似组织上进行了表征。通过对从猫皮质灰质获得的蛋白质材料进行亲和层析分离出GABAA受体。通过SDS-PAGE对抗体进行表征,随后进行免疫印迹和免疫吸附。用MAb进行免疫细胞化学染色显示,在猫视觉皮层的所有层以及白质内均有标记细胞。白质中大多数染色细胞以及皮质灰质中一些细胞的形态与星形胶质细胞非常相似。用抗胶质纤维酸性蛋白(GFAP)抗血清进行双重免疫细胞化学染色,随后进行光学显微镜检查,确实证实猫皮层白质和灰质中的星形胶质细胞都具有GABAA受体。

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