Muscarinic Receptor Knockout Mice Confirm Involvement of M3 Receptor in Endothelium-dependent Vasodilatation in Mouse Arteries

The aim of the study was to determine which cholinergic muscarinic receptor subtype is responsible for the endothelium-dependent vasodilatation evoked by acetylcholine (ACh) in mouse arteries. Endothelium-dependent relaxations were evaluated using isometric tension measurement of ring from femoral and aortic artery of M1, M2, and M3 knockout (KO) mice. Rings of femoral and aortic artery from M3 KO mice did not exhibit relaxation at the opposite of rings from M1+M2 KO and wild-type (WT) mice, which were relaxed by ACh. The proportion of endothelial cells responsive to ACh, as manifested by an increase in cytosolic free calcium ([Ca]i), was also observed on the intima of aorta wall in vitro by using laser line confocal microscopy. Of the cells from M3 KO mice and M1+M2 KO mice, 4% and 23%, respectively, responded to ACh in comparison with 20 % in WT mice. These results show that in the endothelium from femoral and aortic artery, the larger proportion of cells that express M3 receptor is responsible for the specificity of the M3 receptor subtype for endothelium-dependent relaxation caused by ACh.