title: Influence of tissue integrity on pharmacological phenotypes of muscarinic acetylcholine receptors in the rat cerebral cortex.
abstract: Distinct pharmacological phenotypes of muscarinic acetylcholine receptors (mAChRs) have been proposed. We compared the pharmacological profiles of mAChRs in intact segments and homogenates of rat cerebral cortex and other tissues by using radioligand binding assays with [(3)H]N-methylscopolamine ([(3)H]NMS). Recombinant M(1) and M(3) mAChRs were also examined. The density of mAChRs detected by [(3)H]NMS binding to rat cerebral cortex segments and homogenates was the same (approximately 1400 fmol/mg tissue protein), but the dissociation constant of [(3)H]NMS was significantly different (1400-1700 pM in segments and 260 pM in homogenates). A wide variation in [(3)H]NMS binding affinity was also observed among the segments of other tissues (ranging from 139 pM in urinary bladder muscle to 1130 pM in the hippocampus). The mAChRs of cerebral cortex were composed of M(1), M(2), M(3), and M(4) subtypes, which showed typical subtype pharmacology in the homogenates. However, in the cortex segments the M(3) subtype showed a low selectivity for M(3) antagonists (darifenacin, solifenacin) and was not distinguished by the M(3) antagonists from the other subtypes. Recombinant M(1) and M(3) mAChRs showed high affinity for [(3)H]NMS and subtype-specific pharmacology for each tested ligand. The present binding study under conditions where tissue integrity was kept demonstrates a wide variation in [(3)H]NMS binding affinity among mAChRs of many rat tissues and the presence of an atypical M(3) phenotype in the cerebral cortex, suggesting that the pharmacological properties of mAChRs are not necessarily constant, rather they may be significantly modified by tissue integrity and tissue type.
authors: Division of Pharmacology, Department of Biochemistry and Bioinformative Sciences, School of Medicine, University of Fukui, Eiheiji, Fukui, Japan.
authors: Abu Syed
authors: AS
authors: Md Anisuzzaman
authors: Atsushi
authors: A
authors: Nishimune
authors: Hatsumi
authors: H
authors: Yoshiki
authors: Junsuke
authors: J
authors: Uwada
authors: Ikunobu
authors: I
authors: Muramatsu
mesh_terms: D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects; D016466:CHO Cells; D002540:Cerebral Cortex / Q000187:drug effects* / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D003627:Data Interpretation, Statistical; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D018727:Muscarinic Antagonists / Q000378:metabolism; D009130:Muscle, Smooth / Q000378:metabolism; D019832:N-Methylscopolamine / Q000378:metabolism; D010641:Phenotype; D051381:Rats; D017208:Rats, Wistar; D043585:Receptor, Muscarinic M2 / Q000378:metabolism; D043587:Receptor, Muscarinic M3 / Q000187:drug effects; D011976:Receptors, Muscarinic / Q000187:drug effects* / Q000378:metabolism
chemical_list: D018727:Muscarinic Antagonists; D043585:Receptor, Muscarinic M2; D043587:Receptor, Muscarinic M3; D011976:Receptors, Muscarinic; D019832:N-Methylscopolamine