M2 receptor activation of nonselective cation channels in smooth muscle cells: calcium and Gi/G(o) requirements

Muscarinic stimulation of fura 2-loaded smooth muscle cells evoked a rapidly inactivating Ca(2+)-activated Cl- current [ICl(Ca)] and a sustained nonselective cation current (Icat) as well as a transient (delta Ca(tran)) and a sustained (delta Ca(sus)) elevation of cytosolic Ca2+ concentration ([Ca2+]i). Caffeine and inositol 1,4,5-trisphosphate induced delta Ca(tran) and ICl(Ca) but not Icat or delta Ca(sus). M2 receptor antagonism blocked muscarinic activation of Icat and delta Ca(sus) but not ICl(Ca) and delta Ca(tran). M3 antagonism blocked activation of ICl(Ca) and Icat and a rise in [Ca2+]i, but application of caffeine with methacholine restored Icat and delta Ca(sus). After depletion of intracellular Ca2+ stores, methacholine failed to induce Icat or a [Ca2+]i increase and, in pertussis toxin-treated cells, ICl(Ca) and delta Ca(tran) but not Icat or delta Ca(sus) were evoked. Anti-G alpha i-1/G alpha i-2 antibodies and anti-G alpha i-3/ G(o) alpha antibodies blocked Icat but did not affect ICl(Ca). Anti-Gq alpha/ G alpha 11 antibodies greatly inhibited ICl(Ca) but did not affect Icat. Activation of M2 receptors leads to the opening of nonselective cation channels through Gi/G(o) proteins in smooth muscle cells, resulting in a sustained rise in [Ca2+]i. Arise in [Ca2+]i is necessary but not sufficient for activation of nonselective cation channels.