title: Calcium influx through L-type channels attenuates skeletal muscle contraction via inhibition of adenylyl cyclases.
abstract: Skeletal muscle contraction is triggered by acetylcholine induced release of Ca(2+) from sarcoplasmic reticulum. Although this signaling pathway is independent of extracellular Ca(2+), L-type voltage-gated calcium channel (Cav) blockers have inotropic effects on frog skeletal muscles which occur by an unknown mechanism. Taking into account that skeletal muscle fiber expresses Ca(+2)-sensitive adenylyl cyclase (AC) isoforms and that cAMP is able to increase skeletal muscle contraction force, we investigated the role of Ca(2+) influx on mouse skeletal muscle contraction and the putative crosstalk between extracellular Ca(2+) and intracellular cAMP signaling pathways. The effects of Cav blockers (verapamil and nifedipine) and extracellular Ca(2+) chelator EGTA were evaluated on isometric contractility of mouse diaphragm muscle under direct electrical stimulus (supramaximal voltage, 2 ms, 0.1 Hz). Production of cAMP was evaluated by radiometric assay while Ca(2+) transients were assessed by confocal microscopy using L6 cells loaded with fluo-4/AM. Ca(2+) channel blockers verapamil and nifedipine had positive inotropic effect, which was mimicked by removal of extracellular Ca(+2) with EGTA or Ca(2+)-free Tyrode. While phosphodiesterase inhibitor IBMX potentiates verapamil positive inotropic effect, it was abolished by AC inhibitors SQ22536 and NYK80. Finally, the inotropic effect of verapamil was associated with increased intracellular cAMP content and mobilization of intracellular Ca(2+), indicating that positive inotropic effects of Ca(2+) blockers depend on cAMP formation. Together, our results show that extracellular Ca(2+) modulates skeletal muscle contraction, through inhibition of Ca(2+)-sensitive AC. The cross-talk between extracellular calcium and cAMP-dependent signaling pathways appears to regulate the extent of skeletal muscle contraction responses.
authors.affiliation: Division of Cellular Pharmacology, Department of Pharmacology, Escola Paulista de Medicina, Universidade Federal de São Paulo, Rua Três de Maio, 100, 04044-020 São Paulo, SP, Brazil.
authors.forename: Francisco Sandro
authors.initials: FS
authors.lastname: Menezes-Rodrigues
authors.forename: Marcelo
authors.initials: M
authors.lastname: Pires-Oliveira
authors.forename: Thiago
authors.initials: T
authors.lastname: Duarte
authors.forename: Edgar Julian
authors.initials: EJ
authors.lastname: Paredes-Gamero
authors.forename: Tiago
authors.initials: T
authors.lastname: Chiavegatti
authors.forename: Rosely Oliveira
authors.initials: RO
authors.lastname: Godinho
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000067956:Adenylyl Cyclase Inhibitors; D000262:Adenylyl Cyclases / Q000502:physiology; D000818:Animals; D002118:Calcium / Q000502:physiology*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D020746:Calcium Channels, L-Type / Q000502:physiology*; D000242:Cyclic AMP / Q000502:physiology; D007537:Isometric Contraction / Q000187:drug effects / Q000502:physiology; D008297:Male; D051379:Mice; D018482:Muscle, Skeletal / Q000187:drug effects / Q000502:physiology*; D009543:Nifedipine / Q000494:pharmacology; D014700:Verapamil / Q000494:pharmacology
chemical_list: D000067956:Adenylyl Cyclase Inhibitors; D002121:Calcium Channel Blockers; D020746:Calcium Channels, L-Type; C017759:9-(tetrahydro-2-furyl)-adenine; D014700:Verapamil; D000242:Cyclic AMP; D000262:Adenylyl Cyclases; D009543:Nifedipine; D000225:Adenine; D002118:Calcium
keywords: Adenylyl cyclase; Calcium channel blocker; Cyclic AMP; Skeletal muscle