In the present study, we have characterized muscarinic receptor subtypes that mediate carbachol-induced Ca
2+ sensitization of contraction in intestinal smooth muscle, using mutant mice lacking M
2 or M
3 muscarinic receptors or both receptor subtypes. In α-toxin-permeabilized muscle strips from wild-type (WT) mice, isometric tension responses to Ca
2+ applied cumulatively (pCa 7.0-5.0) were increased when the muscarinic agonist carbachol (100 μM) was added to the medium, as judged from shifts of pCa-tension curves in both 50% effective concentration (EC
50) and maximum response (E
max) of pCa-tension curve. In preparations from M
2-knockout (KO) mice, pCa-tension curves were also shifted by carbachol (100 μM), and the extents of the EC
50 and E
max changes resembled those observed in preparations from WT mice. In preparations from M
3-KO or M
2/M
3-double KO mice, however, no significant changes in pCa-tension curves were obtained after carbachol application. The G
q/11-type G-protein inhibitor YM-254890 (1 μM) completely blocked the Ca
2+ sensitization of contraction induced by carbachol in M
2-KO or WT preparations. The results strongly support the idea that the muscarinic activation of Ca
2+ sensitization in intestinal smooth muscles is mediated by the M
3 muscarinic receptor coupled to G
q/11-type G-proteins, without any significant involvement of the other muscarinic receptor subtypes including M
2.
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