Phorbol esters inhibit the synthesis of acetylcholine receptors in cultured muscle cells.
The acetylcholine receptor (AChR) synthesis, insertion and degradation rates are regulated by numerous intracellular and extracellular agents. Recent studies have shown that Ca2+ and Ca2+ ionophores have a profound regulatory effect on the appearance of AChR clusters and AChR synthesis. These regulatory effects may be mediated through the activation of calcium and phospholipid-dependent protein kinases by agents such as phorbol esters. In this study, we have utilized 4-beta-phorbol-12-myristate-13-acetate (PMA) in order to determine whether the activation of protein kinase C exerts a regulatory effect on the expression of AChRs in cultured chick myotubes. Our results show that 4-beta-phorbol-12-myristate-13-acetate decreased intracellular AChRs and suppressed AChR synthesis without affecting the turnover rate. Control and PMA treated cells labeled with [35S] methionine and immunoprecipitated with a monoclonal antibody to the alpha subunit of AChRs (mAb35) revealed a significant decrease in radioactivity precipitated after exposure to PMA. Polyacrylamide gel electrophoresis revealed no major changes in protein patterns, or in newly synthesized proteins as determined by [35S] methionine incorporation and autoradiography. Other enzymes important in muscle metabolism were not affected by PMA treatment. Our results indicate that activation of protein kinase C results in the suppression of AChRs synthesis and dispersal of AChR clusters.
Bursztajn
S
S
Department of Neurology, Baylor College of Medicine, Houston, TX 77030.
Schneider
L W
LW
Jong
Y J
YJ
Berman
S A
SA
D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D003864:Depression, Chemical; D009132:Muscles / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D010703:Phorbol Esters / Q000494:pharmacology*; D011950:Receptors, Cholinergic / Q000096:biosynthesis* / Q000187:drug effects; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D010703:Phorbol Esters; D011950:Receptors, Cholinergic; D013755:Tetradecanoylphorbol Acetate