Clathrin-coated vesicles from bovine brain contain uncoupled GABAA receptors

Brain Research
M H TehraniE M Barnes

Abstract

Clathrin-coated vesicles are thought to be a vehicle for the sequestration of GABAA receptors. For coated vesicles from bovine cerebrum, we examined the binding properties of [3H]muscimol. a GABAA-specific agonist. [3H]flunitrazepam a benzodiazepine agonist, and [35S]t-butylbiocyclophosphorthionate (TBPS), a ligand for GABAA receptor channels. Under standard conditions, the binding level of [3H]muscimol, [3H]flunitrazepam, and [35S]TBPS to coated vesicles represented 12.3 +/- 1.8%, 7.9 +/- 1%, and 10.2 +/- 1.8%, respectively, of that in crude synaptic membranes. Coated vesicles showed a single [3H]flunitrazepam binding site with a KD value (12 nM) which was 9-fold that for synaptic membranes. The allosteric coupling between binding sites was measured by the addition of GABA to [3H]flunitrazepam and [35S]TBPS binding assays. For [3H]flunitrazepam binding to synaptic membranes, GABA gave an EC50 = 2.0 microM and at saturation (100 microM) an enhancement of 122%. This stimulation was completely blocked by the GABA antagonist SR95531. In contrast, neither GABA nor SR95531 had a significant effect on [3H]flunitrazepam binding to CCVs, indicating that the allosteric interaction between GABA and benzodiazepine binding sites is abolish...Continue Reading

References

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Apr 18, 1997·Brain Research. Developmental Brain Research·J D MirandaE M Barnes

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Citations

Dec 1, 1998·European Journal of Pharmacology·M Okubo, M Kawaguchi
Mar 29, 2000·Life Sciences·E M Barnes
Jan 7, 2016·Pharmacological Research : the Official Journal of the Italian Pharmacological Society·María Clara Gravielle
Aug 14, 2009·The FEBS Journal·Jeannine MohrlüderDieter Willbold
Dec 17, 2009·Journal of Neurochemistry·Kelly J SkilbeckTina Hinton
Jun 17, 2010·The European Journal of Neuroscience·Alex M Thomson, Jasmina N Jovanovic
Jul 13, 2004·Pharmacology & Therapeutics·Bernhard Lüscher, Cheryl A Keller
May 30, 2002·The Journal of Biological Chemistry·Mike BeckF Anne Stephenson

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