Calcitonin gene-related peptide potentiates nicotinic acetylcholine receptor-operated slow Ca2+ mobilization at mouse muscle endplates.

1. The involvement of calcitonin gene-related peptide (CGRP) in the non-contractile slow Ca2+ mobilization induced by prolonged nicotinic stimulation was investigated by measurement of [Ca2+], levels in mouse single muscle cells (flexor digitorum brevis; FDB) loaded with a Ca2+ indicator fluo-3 using confocal laser scanning microscopy. 2. CGRP (3-30 nM) potentiated acetylcholine (ACh, 1 microM)-elicited slow Ca2+ mobilization in a concentration-dependent manner. 3. The potentiation by CGRP of the slow Ca2+ component was greatly depressed by a competitive nicotinic antagonist (+)-tubocurarine (5 microM). The Ca2+ channel blocker nitrendipine (1 microM) affected neither ACh responses nor the CGRP potentiation. 4. The slow Ca2+ component was completely abolished by reducing [Ca2+]0 from 2.5 to 0.25 mM whereas the fast component was not affected. The CGRP-induced potentiation of slow Ca2+ signal was also depressed by decreasing [Ca2+]0. 5. Isoproterenol (30 microM) and 8-bromo-adenosine 3',5'-cyclic monophosphate (1 mM) potentiated the ACh-elicited slow Ca2+ response. The potentiation by CGRP of the slow Ca2+ component was completely abolished by a protein kinase-A inhibitor H-89 (1 microM). 6. These findings indicate that CGRP potentiates the nicotinic ACh receptor-operated slow Ca2+ signal via the activation of protein kinase-A system at the skeletal muscle endplates.