title: Involvement of the dihydropyridine receptor and internal Ca2+ stores in myoblast fusion.
abstract: The process of myoblast fusion during skeletal myogenesis is calcium regulated. Both dihydropyridine receptor and ryanodine receptor are already present on muscle precursors, at the prefusional stage, before they are required for excitation-contraction coupling. Previous pharmacological studies have shown the need for a special pool of Ca2+ associated with the membrane for the fusion process to occur. We hypothesized that this pool of Ca2+ is mobilized via a machinery similar to that involved in excitation-contraction coupling. The process of fusion in rat L6 muscle precursors was either totally or partially abolished in the presence of the L-type calcium channel inhibitors SR33557 and nifedipine (half inhibition towards 2 microM), respectively. The inhibition was reversible and dose-dependent. Drugs able to deplete internal calcium stores (caffeine, ryanodine, and thapsigargin) were also tested on the fusion. Both caffeine and thapsigargin drastically inhibited fusion whereas ryanodine had no effect. This suggests that fusion may be controlled by internal pools of Ca2+ but that its regulation may be insensitive to ryanodine. We presumed that an early form of the ryanodine receptor may exist, with different pharmacological properties than the adult forms. Indeed, Western blot analysis of pre- and postfusional L6 cells demonstrated the presence, at the prefusional stage, of a transient form of the ryanodine receptor protein with an apparent molecular weight slightly different from those of the classical skeletal and cardiac forms. Taken together, these results support the hypothesis that the fusion process is driven by a mechanism involving both the dihydropyridine receptor (alpha1 subunit of the L-type Ca2+ channel) and the internal stores of Ca2+. The machinery underlying this mechanism might consist of slightly different forms of the classic molecules that in adult muscle ensure excitation-contraction coupling. It remains to be seen, however, whether the mobilization of the internal pool of Ca2+ is triggered by the type of mechanism already described in skeletal muscle.
authors.affiliation: INSERM U153, Paris, France.
authors.forename: S
authors.initials: S
authors.lastname: Seigneurin-Venin
authors.forename: E
authors.initials: E
authors.lastname: Parrish
authors.forename: I
authors.initials: I
authors.lastname: Marty
authors.forename: F
authors.initials: F
authors.lastname: Rieger
authors.forename: G
authors.initials: G
authors.lastname: Romey
authors.forename: M
authors.initials: M
authors.lastname: Villaz
authors.forename: L
authors.initials: L
authors.lastname: Garcia
mesh_terms: D000818:Animals; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D015220:Calcium Channels / Q000032:analysis / Q000502:physiology*; D020746:Calcium Channels, L-Type; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors; D002459:Cell Fusion / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D003402:Creatine Kinase / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007212:Indolizines / Q000494:pharmacology; D015640:Ion Channel Gating; D018485:Muscle Fibers, Skeletal / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D009124:Muscle Proteins / Q000032:analysis / Q000502:physiology*; D009132:Muscles / Q000196:embryology; D009543:Nifedipine / Q000494:pharmacology; D010627:Phenethylamines / Q000494:pharmacology; D051381:Rats; D012433:Ryanodine / Q000494:pharmacology; D019837:Ryanodine Receptor Calcium Release Channel; D013729:Terpenes / Q000494:pharmacology; D019284:Thapsigargin
chemical_list: D002121:Calcium Channel Blockers; D015220:Calcium Channels; D020746:Calcium Channels, L-Type; D004791:Enzyme Inhibitors; D007212:Indolizines; D009124:Muscle Proteins; D010627:Phenethylamines; D019837:Ryanodine Receptor Calcium Release Channel; D013729:Terpenes; D012433:Ryanodine; D002110:Caffeine; D019284:Thapsigargin; D003402:Creatine Kinase; D000252:Calcium-Transporting ATPases; D009543:Nifedipine; C059901:fantofarone; D002118:Calcium