Immunocytochemical detection of astrocyte GABAA receptors in cat visual cortex

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.