Ractopamine increases total and myofibrillar protein synthesis in cultured rat myotubes.
The ability of the phenethanolamine (beta-adrenergic agonist) ractopamine to stimulate cellular protein accretion and protein synthesis in cultured muscle cells was evaluated. ELC5 myoblasts (a subclone of rat L6 cells) were proliferated in culture (Dulbecco's Modified Eagle Medium plus 10% fetal bovine serum at 37 degrees C) to confluency and then allowed to differentiate to form myotubes. Myotubes were then further incubated in the presence of 10(-9), 10(-8), 10(-7), 10(-6) or 10(-5) mol/L ractopamine. A significant (p less than 0.05) response in cellular protein accretion was observed for the 10(-6) and 10(-5) concentrations when compared to 10(-8) and 10(-9) mol/L ractopamine. Ractopamine at 0 and 10(-6) mol/L was used to examine the effect of the beta agonist on [35S]methionine incorporation (protein synthesis) into total cellular protein, 43-kDa proteins and myosin heavy-chain (200 kDa) protein. Protein synthesis in response to beta agonist treatment was measured at 4, 24, 48, 72 and 96 h after ractopamine addition to the ELC5 myotubes in culture. Ractopamine (10(-6) mol/L) increased [35S]methionine incorporation (apparent protein synthesis) at 24 h (p less than 0.01), 48 h (p less than 0.05), 72 h (p less than 0.01) and 96 h (p less than 0.05) in cultured ELC5 muscle cells. Ractopamine also increased apparent protein synthesis rate of the 43-kDa proteins (p less than 0.05) and myosin heavy-chain protein (200 kDa) (p less than 0.05). These results indicate that ractopamine-enhanced ELC5 myotube protein accretion is mediated, at least in part, by stimulating cellular protein synthesis.
Anderson
P T
PT
Department of Animal Science, Michigan State University, East Lansing 48824.
Helferich
W G
WG
Parkhill
L C
LC
Merkel
R A
RA
Bergen
W G
WG
D000318:Adrenergic beta-Agonists / Q000494:pharmacology*; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D008970:Molecular Weight; D009124:Muscle Proteins / Q000096:biosynthesis*; D009132:Muscles / Q000187:drug effects / Q000378:metabolism*; D009210:Myofibrils / Q000378:metabolism*; D009218:Myosins / Q000096:biosynthesis; D010627:Phenethylamines / Q000494:pharmacology*; D051381:Rats
D000318:Adrenergic beta-Agonists; D009124:Muscle Proteins; D010627:Phenethylamines; C060897:ractopamine; D009218:Myosins