Differential modulation of CaV2.3 Ca2+ channels by Galphaq/11-coupled muscarinic receptors.
CaV2.3 subunits are expressed in neuronal and neuroendocrine cells where they are believed to form native R-type Ca2+ channels. Although R-type currents are involved in triggering neurotransmitter and hormone secretion, little is known about their modulation. Previous studies have shown that muscarinic acetylcholine receptors evoke both inhibition and stimulation of CaV2.3. Muscarinic inhibition of CaV2.3 is mediated by Gbetagamma subunits, whereas stimulation is mediated by pertussis toxin-insensitive Galpha subunits. In the present study, we compared modulation of CaV2.3 by the three Galphaq/11-coupled muscarinic receptors (M1, M3, and M5). Our data indicate that these receptors trigger comparable stimulation of CaV2.3. The signaling pathway that mediates stimulation was meticulously analyzed for M1 receptors. Stimulation is blocked by neutralizing antibodies directed against Galphaq/11, coexpression of the regulatory domain of protein kinase Cdelta (PKCdelta), preactivating PKC with phorbol ester, or pharmacological suppression of PKC with bisindolylmaleimide I. Stimulation of CaV2.3 is Ca(2+)-independent and insensitive to 12-(2-cyanoethyl)-6,7,12,13-tetrahydro-13-methyl-5-oxo-5H-indolo(2,3-a)pyrrolo(3,4-c)-carbazole (Gö 6976), a specific inhibitor of Ca(2+)-dependent PKC isozymes. These results indicate that muscarinic stimulation of CaV2.3 involves signaling by Galphaq/11, diacylglycerol, and a Ca(2+)-independent PKC. In contrast to stimulation, the magnitude of CaV2.3 inhibition depended on receptor subtype, with M3 and M5 receptors producing much larger CaV2.3 inhibition than M1 receptors. Interestingly, muscarinic inhibition of CaV2.3 was notably enhanced during pharmacological suppression of PKC, suggesting the presence of cross-talk between Gbetagamma-mediated inhibition and PKC-mediated stimulation of R-type channels similar to that described previously for N-type channels.
Bannister
R A
RA
Department of Biology, Utah State University, Logan, Utah 84322, USA.
Melliti
K
K
Adams
B A
BA
D000906:Antibodies / Q000494:pharmacology; D020908:Calcium Channels, R-Type / Q000378:metabolism*; D002460:Cell Line; D004791:Enzyme Inhibitors / Q000494:pharmacology; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000502:physiology*; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D018408:Patch-Clamp Techniques; D043562:Receptors, G-Protein-Coupled / Q000502:physiology; D011976:Receptors, Muscarinic / Q000502:physiology*
D000906:Antibodies; D020908:Calcium Channels, R-Type; D004791:Enzyme Inhibitors; D043562:Receptors, G-Protein-Coupled; D011976:Receptors, Muscarinic; D043802:GTP-Binding Protein alpha Subunits, Gq-G11