Regional and cellular expression sites of the alpha 1 subunit of GABAA receptors in the rat basal forebrain: a cytochemical study with glutamic acid decarboxylase, choline acetyltransferase, calcium-binding proteins and nitric oxide synthase as second markers

Abstract

Forebrain sections of adult male Wistar rats were processed for the immunohistochemical detection of the GABAA receptor alpha 1 subunit. Alternate sections were used for double-staining with antibodies to glutamic acid decarboxylase (GAD), choline acetyltransferase (ChAT), the calcium binding proteins parvalbumin (PARV), calbindin (CALB) and calretinin (CR) as well as to nitric oxide synthase (NOS). alpha 1 receptor subunit-immunoreactive neurons were found to be inhomogeneously distributed in the rat basal forebrain. Numerous alpha 1 subunit-immunostained neurons occupied the central part of medial septum and diagonal band, the whole ventral pallidum and the globus pallidus. A moderate number was found in the lateral septum, and only very few in the striatum and nucleus accumbens. Double-immunofluorescence labelling revealed an apparently complete co-expression of GAD-immunoreactivity in alpha 1 subunit-immunoreactive cells of rat basal forebrain, but only a region-dependent proportion of GAD-immunoreactive cells showed alpha 1 subunit-immunoreactivity. Co-expression of PARV-immunoreactivity characterized the vast majority of the alpha 1 subunit-immunoreactive cells in the medial septum, diagonal band, ventral pallidum and globus pallidus. Striatal alpha 1 subunit-immunopositive neurons appeared PARV-immunonegative and did also not react with the other immunoreagents used in this study, except the GAD-antibody. CR-immunoreactivity was co-expressed in alpha 1 subunit-immunopositive cells of the ventral lateral septal nucleus and only exceptionally in the ventral pallidum, where the vast majority of CR-positive cells was monolabelled. A small minority of ChAT-immunoreactive, but in no case CALB- and NOS-immunoreactive cells were found to express the alpha 1 subunit-immunoreactivity. These findings confirm the data obtained by analyses of other brain regions suggesting a preferred co-existence of this GABAA receptor subunit with PARV and to a lesser degree with CR.