Expression of type I and III collagen genes during differentiation of embryonic chicken myoblasts in culture.
Expression of type I and III procollagen genes was studied in embryonic chicken myoblast cell cultures, obtained from thigh muscles of 11-day-old embryos. Differentiation initiated by the addition of ovotransferrin (30 micrograms/ml) was followed visually by phase-contrast microscopy. Myoblast fusion and myotube formation were detected by day 3 and appeared to be complete by day 7. The synthesis of procollagens was monitored by labeling cell cultures for 1 h with [3H]proline and determining the radioactivity in procollagen chains by scanning densitometry of the fluorograms of the sodium dodecyl sulfate-polyacrylamide gels. A 10- to 20-fold increase in the rate of pro alpha-1(I), pro alpha-2(I), and pro alpha-1(III) collagen synthesis was observed, with the greatest increase occurring between days 3 and 9. Collagen mRNA levels in the myoblast cultures were examined by Northern blot and dot blot hybridization assays. The 10- to 20-fold increased rate of protein synthesis was accompanied by a 15-fold increase in the steady-state levels of pro alpha-1(I) and pro alpha-2(I) mRNAs and a 10-fold increase in the steady-state levels of pro alpha-1(III). As a correlate to the studies of collagen expression during myoblast differentiation, the expression of actin mRNAs was examined. Although alpha actin could be detected by day 4, a complete switch from lambda and beta to alpha actin was not observed in the time periods examined. Similar results were obtained in the analysis of RNA extracted from embryonic legs at days 12 and 17 of gestation. Myoblast differentiation is manifested by the accumulation of both muscle-specific mRNAs, such as actin, and type I and III procollagen mRNAs.
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002642:Chick Embryo; D003001:Cloning, Molecular; D003094:Collagen / Q000235:genetics*; D003207:Conalbumin / Q000494:pharmacology; D004247:DNA / Q000032:analysis; D005796:Genes; D007700:Kinetics; D009132:Muscles / Q000166:cytology / Q000187:drug effects / Q000196:embryology*; D009693:Nucleic Acid Hybridization; D011347:Procollagen / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification
D011347:Procollagen; D012333:RNA, Messenger; D003207:Conalbumin; D003094:Collagen; D004247:DNA