Bidirectional regulation of GABAA receptor alpha1 and alpha6 subunit expression by a cyclic AMP-mediated signalling mechanism in cerebellar granule cells in primary culture.
Forskolin treatment of cerebellar granule cells in culture resulted in bidirectional regulation of the expression of GABAA receptor alpha1 and alpha6 subunits. Thus, forskolin applied at 2 days in vitro (DIV) increased expression of the alpha1 subunit but decreased the expression of the alpha6 subunit. Values with respect to control cultures, both assayed at 9 DIV by immunoblotting, were 310 +/- 48% for alpha1 and 25 +/- 16% for the alpha6 subunit. Similar effects were evoked following chronic treatment with both dibutyryl cyclic AMP and 3-isobutyl-1-methylxanthine. Dideoxyforskolin had no effect on the level of expression of either the alpha1 or the alpha6 GABAA receptor subunits. The changes in subunit expression were accompanied by a 1.7-fold increase in number of total specific [3H]Ro 15-4513 binding sites expressed by intact cerebellar granule cells. This increase in total binding sites was accommodated by a 2.7-fold increase in number of diazepam-sensitive Ro 15-4513 binding sites in accordance with the observed increase in alpha1 subunit expression. The number of diazepam-insensitive subtype of binding sites were not significantly changed. These results suggest that GABAA receptor subtype expression can be differentially regulated by intracellular cyclic AMP concentration.
Thompson
C L
CL
Department of Pharmaceutical and Biological Chemistry, School of Pharmacy, University of London, England.
Pollard
S
S
Stephenson
F A
FA
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000345:Affinity Labels; D000818:Animals; D001386:Azides / Q000494:pharmacology; D001569:Benzodiazepines / Q000494:pharmacology; D003994:Bucladesine / Q000494:pharmacology; D002478:Cells, Cultured / Q000737:chemistry / Q000502:physiology; D002531:Cerebellum / Q000166:cytology; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000502:physiology*; D009474:Neurons / Q000737:chemistry / Q000166:cytology / Q000502:physiology*; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D011869:Radioligand Assay; D051381:Rats; D017208:Rats, Wistar; D011963:Receptors, GABA-A / Q000096:biosynthesis / Q000737:chemistry / Q000502:physiology*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D014316:Tritium / Q000378:metabolism
D000345:Affinity Labels; D001386:Azides; D010726:Phosphodiesterase Inhibitors; D011963:Receptors, GABA-A; D014316:Tritium; D001569:Benzodiazepines; D005576:Colforsin; D003994:Bucladesine; C042957:Ro 15-4513; D000242:Cyclic AMP; D015056:1-Methyl-3-isobutylxanthine