Testosterone-induced hypertrophy of L6 myoblasts is dependent upon Erk and mTOR.
Testosterone increases the size and strength of skeletal muscle. This study further characterized the molecular mechanisms of the anabolic actions of testosterone on a rat myoblast cell line (L6 cells). Testosterone did not induce hypertrophy in L6 cells lacking the androgen receptor (AR). Hypertrophy was prevented by the AR antagonist bicalutamide and the mTOR inhibitor rapamycin. Testosterone induced Erk phosphorylation by 2h, and mTOR autophosphorylation was elevated within 20min; phosphorylation of p70S6 kinase was increased by 2h. Inhibitors of Erk or PI3K blocked tesotosterone-induced hypertrophy. Erk phosphorylation returned to baseline when media containing testosterone was replaced at 16h with fresh media lacking testosterone; when bicalutamide was added to testosterone-enriched media at 16h, Erk phosphorylation remained elevated. Autophosphorylation of the IGF-1 receptor was minimally altered by testosterone at 20min and unaffected at later time points; PI3K/PDK1-dependent phosphorylation of Akt was not altered by testosterone. These findings indicate that testosterone stimulates hypertrophy of L6 myoblasts through a mechanism that requires its binding to the AR and involves a signaling cascade dependent upon Erk and mTOR which is likely activated by substances released into the extracellular space which are not IGF-1 or other ligands for receptor tyrosine kinases.
Wu
Yong
Y
Department of Veterans Affairs, Center of Excellence for the Medical Consequences of Spinal Cord Injury, Room 1E-02, James J. Peters VA Medical Center, West Kingsbridge Road, Bronx, NY 10468, USA.
Bauman
William A
WA
Blitzer
Robert D
RD
Cardozo
Christopher
C
D000818:Animals; D049108:Cell Enlargement; D002460:Cell Line; D048049:Extracellular Signal-Regulated MAP Kinases / Q000235:genetics / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D032446:Myoblasts / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D010766:Phosphorylation; D051381:Rats; D011944:Receptors, Androgen / Q000235:genetics / Q000502:physiology*; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D013739:Testosterone / Q000494:pharmacology / Q000502:physiology*
D011944:Receptors, Androgen; D013739:Testosterone; C546845:mTOR protein, rat; D058570:TOR Serine-Threonine Kinases; D048049:Extracellular Signal-Regulated MAP Kinases