Increased endocytosis of acetylcholine receptors by dystrophic mouse myotubes in vitro.
Multinucleated myotubes, grown in vitro from satellite cells of dystrophic mice (C57BL/6J/dydy) exhibit a reduced sensitivity to ACh. This reduction correlates with a reduced density of 125I-alpha-bungarotoxin (125I-BTX) binding sites on the surface of dystrophic myotubes. Denervated adult muscle fibers from dystrophic mice respond to Ach similarly to denervated normal muscle fibers. Furthermore, cultured dystrophic myotubes, treated with a brain extract which induces AChR clusterization, still show an impaired response to ACh and reduced 125I-BTX binding. Thus AChR function appears altered in dystrophic muscle cells in culture while it appears normal in dystrophic adult muscle, regardless of whether the receptors are dispersed on the membrane or clustered at the junctional site. Metabolic studies on the reduced AChR level in dystrophic myotubes revealed a dramatically reduced half-life (2 vs 10 hr) while the rate of synthesis was unchanged. An increased rate of internalization of AChR was observed in dystrophic myotubes with a corresponding relative increase of the "hidden AChR pool," which could be partially reduced by agents which disrupt the cytoskeleton. No structural alterations could be detected on the AChR molecule as its sedimentation coefficient and subunit composition appeared identical between normal and dystrophic myotubes. Thus the increased turnover of AChR in dystrophic myotubes either reflects subtle alterations of the molecule or a more generalized increase of endocytosis in this form of myopathy.
D000818:Animals; D001696:Biomechanical Phenomena; D002478:Cells, Cultured; D002738:Chloroquine / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004705:Endocytosis; D006614:Hindlimb; D008715:Methionine; D051379:Mice; D008810:Mice, Inbred C57BL; D009121:Muscle Denervation; D009132:Muscles / Q000166:cytology / Q000378:metabolism*; D009136:Muscular Dystrophies / Q000378:metabolism*; D011950:Receptors, Cholinergic / Q000145:classification / Q000378:metabolism*; D014357:Trypsin / Q000494:pharmacology
D011950:Receptors, Cholinergic; D003571:Cytochalasin B; D002738:Chloroquine; D008715:Methionine; D014357:Trypsin