mRNA levels of cathepsins B and D during myogenesis.
Muscle development is characterized by the fusion of myoblasts to form myotubes and the co-ordinate expression of muscle-specific proteins such as actin, myosin and creatine phosphokinase. Our laboratory has been involved in the study of the role of lysosomal proteinases, namely, cathepsins B, H and L and the endogenous cysteine proteinase inhibitor, cystatin, during muscle differentiation in vitro. Specific activities of the cysteine proteinase in chicken primary cultures and a number of rat myogenic lines increased with the degree of myotube formation. This has suggested that lysosomal proteinases play an important role in myogenesis. We have measured the mRNA levels of two of the lysosomal enzymes, cathepsins B and D. This is advantageous because the presence of the endogenous inhibitor, cystatin, masks the levels of the specific activities of the cysteine proteinases present in the cell. RNA was extracted from developing muscle at three stages of development: proliferating myoblasts, confluent cells, and myotubes. Hybridization of RNA extracted from the L6 myogenic cell line with cathepsin B cDNA showed an increase in the level of cathepsin B mRNA. However, in the L8 rat myogenic line the level decreased after fusion. Cathepsin D mRNA levels remained constant throughout differentiation of the L8 cells. This paper also reports on the characterization of lysosomal proteinases of a newly obtained mouse myogenic line, C2.
D000818:Animals; D002401:Cathepsin B / Q000096:biosynthesis*; D002402:Cathepsin D / Q000096:biosynthesis*; D002460:Cell Line; D004247:DNA / Q000096:biosynthesis; D008247:Lysosomes / Q000201:enzymology; D024510:Muscle Development; D009132:Muscles / Q000201:enzymology; D009693:Nucleic Acid Hybridization; D010447:Peptide Hydrolases / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats
D012333:RNA, Messenger; D004247:DNA; D010447:Peptide Hydrolases; D002401:Cathepsin B; D002402:Cathepsin D