title: Uncoupling of M1 muscarinic receptor/G-protein interaction by amyloid β(1-42).
abstract: The overproduction of β-amyloid (Aβ) fragments in transgenic APPswe/PS1dE9 mice results in formation of amyloid deposits in the cerebral cortex and hippocampus starting around four months of age and leading to cognitive impairment much later. We have previously found an age and transgene-dependent weakening of muscarinic receptor-mediated transmission that was not present in young (6-10-week-old) animals but preceded both amyloid deposits and cognitive deficits. Now we investigated immediate and prolonged in vitro effects of non-aggregated Aβ(1-42) on coupling of individual muscarinic receptor subtypes expressed in CHO (Chinese hamster ovary) cells and their underlying mechanisms. Immediate application of 1 μM Aβ(1-42) had no effect on the binding of the muscarinic antagonist N-methylscopolamine or the agonist carbachol. In contrast, 4-day treatment of CHO cells expressing the M1 muscarinic receptor with 100 nM Aβ(1-42) significantly changed the binding characteristics of the muscarinic agonist carbachol and reduced the extent of the M1 receptor-stimulated breakdown of phosphatidylinositol while it did not demonstrate overt toxic effects. The treatment had no influence on the expression of either G-proteins or muscarinic receptors. In concert, we found no change in the gene expression of muscarinic receptor subtypes and gene or protein expression of the G(s), G(q/11), and G(i/o) G-proteins in the cerebral cortex of young adult APPswe/PS1dE9 mice that demonstrate high concentrations of soluble Aβ(1-42) and impaired muscarinic receptor-mediated G-protein activation. Our results provide strong evidence that the initial injurious effects of Aβ(1-42) on M1 muscarinic receptor-mediated transmissionis is due to compromised coupling of the receptor with G(q/11) G-protein.
authors.affiliation: Institute of Physiology CAS, Vídeňská 1083, 14220 Prague 4, Czech Republic. smyckova@biomed.cas.cz
authors.forename: Helena
authors.initials: H
authors.lastname: Janíčková
authors.forename: Vladimír
authors.initials: V
authors.lastname: Rudajev
authors.forename: Pavel
authors.initials: P
authors.lastname: Zimčík
authors.forename: Jan
authors.initials: J
authors.lastname: Jakubík
authors.forename: Heikki
authors.initials: H
authors.lastname: Tanila
authors.forename: Esam E
authors.initials: EE
authors.lastname: El-Fakahany
authors.forename: Vladimír
authors.initials: V
authors.lastname: Doležal
mesh_terms: D016229:Amyloid beta-Peptides / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D010446:Peptide Fragments / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D043643:Receptor, Muscarinic M1 / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000037:antagonists & inhibitors / Q000378:metabolism*; D014475:Uncoupling Agents / Q000494:pharmacology
chemical_list: D016229:Amyloid beta-Peptides; D010446:Peptide Fragments; D043643:Receptor, Muscarinic M1; D043562:Receptors, G-Protein-Coupled; D014475:Uncoupling Agents; C075222:amyloid beta-protein (1-42)