Effect of atrophy and contractions on myogenin mRNA concentration in chick and rat myoblast omega muscle cells.
The skeletal rat myoblast omega (RMo) cell line forms myotubes that exhibit spontaneous contractions under appropriate conditions in culture. We examined if the RMo cells would provide a model for studying atrophy and muscle contraction. To better understand how to obtain contractile cultures, we examined levels of contraction under different growing conditions. The proliferation medium and density of plating affected the subsequent proportion of spontaneously contracting myotubes. Using a ribonuclease protection assay, we found that exponentially growing RMo myoblasts contained no detectable myogenin or herculin mRNA, while differentiating myoblasts contained high levels of myogenin mRNA but no herculin mRNA. There was no increase in myogenin mRNA concentration in either primary chick or RMo myotubes whose contractions were inhibited by depolarizing concentrations of potassium (K+). Thus, altered myogenin mRNA concentrations are not involved in atrophy of chick myotubes. Depolarizing concentrations of potassium inhibited spontaneous contractions in both RMo cultures and primary chick myotube cultures. However, we found that the myosin concentration of 6-d-old contracting RMo cells fed medium plus AraC was 11 +/- 3 micrograms myosin/microgram DNA, not significantly different from 12 +/- 4 micrograms myosin/microgram DNA (n = 3), the myosin concentration of noncontracting RMo cells (treated with 12 mM K+ for 6 d). Resolving how RMo cells maintained their myosin content when contraction is inhibited may be important for understanding atrophy.
Krebs
J M
JM
University Space Research Association, Johnson Space Center, Houston, Texas 77058, USA.
Denney
R M
RM
D000818:Animals; D002452:Cell Count; D002461:Cell Line, Transformed; D002642:Chick Embryo; D003470:Culture Media; D008954:Models, Biological; D009119:Muscle Contraction / Q000187:drug effects / Q000502:physiology*; D009132:Muscles / Q000196:embryology*; D009133:Muscular Atrophy; D018008:Myogenin / Q000235:genetics*; D009218:Myosins / Q000378:metabolism; D011188:Potassium / Q000008:administration & dosage / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
D003470:Culture Media; C503803:Myog protein, rat; D018008:Myogenin; D012333:RNA, Messenger; D009218:Myosins; D011188:Potassium