Selective inhibition of cathepsin B with cell-permeable CA074Me negatively affects L6 rat myoblast differentiation.
Active cathepsin B, in concert with other cellular proteases, has been implicated in the catabolic restructuring associated with myotube formation during skeletal myoblast cell differentiation (i.e., myogenesis). We have examined this role in differentiating myoblasts using the cell-permeable, cathepsin B selective inhibitor CA074Me. Cathepsin B activity levels in differentiating L6 rat myoblasts treated with CA074Me were significantly lower than levels in control myoblasts. Inhibition of cathepsin B activity by CA074Me occurred at each stage of differentiation and was dose related. Myotube size and number and induced levels of fusion-related creatine phosphokinase activity and myosin heavy-chain protein were reduced from 30 to 50% in CA074Me-treated myoblasts. These reductions were also dose related. In contrast, CA074Me did not affect levels of myogenin, an early marker of myogenesis, or levels of cathepsin L type and myokinase activities, two nonspecific enzymes. The negative effects associated with CA074Me were reversed when the drug was removed. Collectively, these data suggest that active cathepsin B plays a role in myoblast-myoblast fusion and consequently may be necessary for the complete expression of those genes associated with the fusion process.
Jane
Derek T
DT
Department of Biological Sciences, University of Windsor, ON, Canada.
Morvay
Leslie C
LC
Allen
Francis
F
Sloane
Bonnie F
BF
Dufresne
Michael J
MJ
D000818:Animals; D015415:Biomarkers; D002401:Cathepsin B / Q000037:antagonists & inhibitors*; D002454:Cell Differentiation / Q000502:physiology*; D002463:Cell Membrane Permeability; D004151:Dipeptides / Q000494:pharmacology*; D018485:Muscle Fibers, Skeletal / Q000502:physiology; D032446:Myoblasts / Q000378:metabolism*; D051381:Rats
D015415:Biomarkers; C400541:CA 074 methyl ester; D004151:Dipeptides; D002401:Cathepsin B