Activation of nicotinic acetylcholine receptors increases the rate of fusion of cultured human myoblasts.
1. Fusion of myogenic cells is important for muscle growth and repair. The aim of this study was to examine the possible involvement of nicotinic acetylcholine receptors (nAChR) in the fusion process of myoblasts derived from postnatal human satellite cells. 2. Acetylcholine-activated currents (ACh currents) were characterized in pure preparations of freshly isolated satellite cells, proliferating myoblasts, myoblasts triggered to fuse and myotubes, using whole-cell and single-channel voltage clamp recordings. Also, the effect of cholinergic agonists on myoblast fusion was tested. 3. No nAChR were observed in freshly isolated satellite cells. nAChR were first observed in proliferating myoblasts, but ACh current densities increased markedly only just before fusion. At that time most mononucleated myoblasts had ACh current densities similar to those of myotubes. ACh channels had similar properties at all stages of myoblast maturation. 4. The fraction of myoblasts that did not fuse under fusion-promoting conditions had no ACh current and thus resembled freshly isolated satellite cells. 5. The rate of myoblast fusion was increased by carbachol, an effect antagonized by alpha-bungarotoxin, curare and decamethonium, but not by atropine, indicating that nAChR were involved. Even though a prolonged exposure to carbachol led to desensitization, a residual ACh current persisted after several days of exposure to the nicotinic agonist. 6. Our observations suggest that nAChR play a role in myoblast fusion and that part of this role is mediated by the flow of ions through open ACh channels.
Krause
R M
RM
Division de Recherche Clinique Neuro-Musculaire, Hôpital Cantonal Universitaire, Genève, Switzerland.
Hamann
M
M
Bader
C R
CR
Liu
J H
JH
Baroffio
A
A
Bernheim
L
L
D002217:Carbachol / Q000037:antagonists & inhibitors / Q000494:pharmacology; D002459:Cell Fusion / Q000187:drug effects; D002478:Cells, Cultured; D004594:Electrophysiology; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism; D018482:Muscle, Skeletal / Q000166:cytology* / Q000187:drug effects; D018722:Nicotinic Agonists / Q000494:pharmacology*; D018408:Patch-Clamp Techniques; D011978:Receptors, Nicotinic / Q000187:drug effects / Q000378:metabolism*
D018722:Nicotinic Agonists; D011978:Receptors, Nicotinic; D002217:Carbachol