Measurement of protein synthesis and degradation in C2C2) myoblasts using extracts of muscle from hormone treated bovine.
A detailed methodology is described for determination of treatment effects on muscle cell protein synthesis and muscle cell protein degradation in a cell culture system. C(2)C(22) mouse myoblasts were treated with growth media containing muscle extracts from bovine treated with different pharmaceutical agents. Radiolabeled amino acids were added to the growth media to determine treatment effects on protein synthesis and protein degradation. Percent protein synthesis was calculated by measuring amino acid uptake as a percentage over internal control. Percent protein degradation was measured using a pulse chase technique. These procedures will allow researchers to determine treatment effects on overall protein synthesis and degradation in vitro in a relatively short amount of time without excessive costs. A second benefit is that animals do not have to be fed radiolabeled feedstuffs. These procedures are not intended to elucidate the mechanisms behind pharmaceutical enhancement of muscle cell protein synthesis or protein degradation.
Montgomery
J L
JL
Department of Animal Science, Texas Tech University, Lubbock, TX 79409, USA. jblanton@ttacs.ttu.edu
Harper
W M
WM
Miller
M F
MF
Morrow
K J
KJ
Blanton
J R
JR
D000596:Amino Acids / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D002250:Carbon Radioisotopes; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D006133:Growth Substances / Q000494:pharmacology; D051379:Mice; D009124:Muscle Proteins / Q000737:chemistry / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D032448:Myoblasts, Skeletal / Q000187:drug effects / Q000378:metabolism*; D014020:Tissue Extracts / Q000494:pharmacology*
D000596:Amino Acids; D002250:Carbon Radioisotopes; D003470:Culture Media; D006133:Growth Substances; D009124:Muscle Proteins; D014020:Tissue Extracts