GABAA Receptor Subtypes Expressed in Cerebellar Granule Cells: A Developmental Study

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Abstract
The developmental properties of primary rat cerebellar granule cells have been characterised with respect to their expression of GABAA receptor subtypes using both an immunological approach and radioligand binding assays. At day 1 in culture, the GABAA receptor alpha 1 subunit was detectable in immunoblots and increased in level up to day 9. The GABAA receptor alpha 6 subunit was not detectable at day 1; however, at days 3-5, a specific M(r) 58,000 anti-alpha 6 1-16 Cys immunoreactive species was present which further increased in level up to 9 days in culture. Similar qualitative results were obtained for the expression of the GABAA receptor alpha 6 subunit in age-matched rat cerebellar membranes. In parallel studies, it was found that although there was an overall increase in [3H]Ro 15-4513 binding sites with days in culture, the relative contributions of diazepam-sensitive and diazepam-insensitive [3H]Ro 15-4513 binding changed. A time-dependent enrichment of the diazepam-insensitive binding site up to a maximum of 74% of total [3H]Ro 15-4513 sites was found. This was concomitant with the appearance of the GABAA receptor alpha 6 subunit. These results are in agreement with the pharmacology described for alpha 6 beta gamma 2 cloned receptors. They suggest a developmentally regulated expression of the GABAA receptor alpha 6 subunit gene at a time that is correlated in vivo with establishment of neuronal connections.