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 Aβ1-42 on coupling of individual muscarinic receptor subtypes expressed in CHO (Chinese hamster ovary) cells and their underlying mechanisms. Immediate application of 1 μM Aβ1-42 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 Gs, Gq/11, and Gi/o G-proteins in the cerebral cortex of young adult APPswe/PS1dE9 mice that demonstrate high concentrations of soluble Aβ1-42 and impaired muscarinic receptor-mediated G-protein activation. Our results provide strong evidence that the initial injurious effects of Aβ1-42 on M1 muscarinic receptor-mediated transmissionis is due to compromised coupling of the receptor with Gq/11 G-protein.
Bibliographical noteFunding Information:
We are grateful to Dr. O. Benada for electron microscopy images and Dr. L. Bacakova for help with light microscopy. This research was supported by projects EU FP7 project LipiDiDiet (Grant Agreement No 211696), AV0Z50110509, RVO: 67985823, and grants GACR 305/09/0681 and P304/12/G069.
- Agonist binding
- Amyloid β
- Muscarinic receptors
- Phosphatidylinositol breakdown