Pharmacological Assessment of M1 Muscarinic Acetylcholine Receptor-Gq/11 Protein Coupling in Membranes Prepared from Postmortem Human Brain Tissue

Using a selective Gαq/11 protein antibody capture guanosine 5′- O -(3-[35S]thio)triphosphate ([35S]GTPγS) binding approach, it has been possible to perform a quantitative pharmacological examination of the functional activity of the M1 muscarinic acetylcholine receptor (mAChR) in membranes prepared from human postmortem cerebral cortex. Oxotremorine-M caused a ≥2-fold increase in [35S]GTPγ S -Gαq/11 binding with a pEC50 of 6.06 ± 0.16 in Brodmann's areas 23 and 25 that was almost completely inhibited by preincubation of membranes with the M1 mAChR subtype-selective antagonist muscarinic toxin-7. In addition, the orthosteric and allosteric agonists, xanomeline [3(3-hexyloxy-1,2,5-thiadiazol-4-yl)-1,2,5,6-tetrahydro-1-methylpyridine] and AC-42 (4- n -butyl-1-[4-(2-methylphenyl)-4-oxo-1-butyl]-piperidine hydrogen chloride), increased [35S]-GTPγ S -Gαq/11 binding, but with reduced intrinsic activities, inducing maximal responses that were 42 ± 1 and 44 ± 2% of the oxotremorine-M-induced response, respectively. These data indicate that the M1 receptor is the predominant mAChR subtype coupling to the Gαq/11 G protein in these brain regions and that it is possible to quantify the potency and intrinsic activity of full and partial M1 mAChR receptor agonists in postmortem human brain using a selective Gαq/11 protein antibody capture [35S]GTPγS binding assay.