Desmin degradation and Ca(2+)-dependent proteolysis during myoblast fusion.
It has already been reported that, in vitro, intermediate filaments such as desmin and vimentin are very susceptible to proteolysis by calpains (Ca(2+)-activated cysteine proteinases). On the other hand, desmin and m-calpain are both present at the onset of myoblast fusion and throughout this phenomenon. Based on these observations, the aim of this study was to demonstrate, with cultured rat myoblasts, that the amount of desmin decreased significantly as multinucleated myotubes were formed. Using immunoblot analysis, it has been shown that the desmin concentration decreased 41% as myoblasts fuse. Moreover, under conditions which stimulate myoblast fusion, desmin concentration was reduced by 21% compared to the control culture. Under our experimental conditions, which lead to a reduced desmin level, the amount of m-calpain was increased about three-fold. These results suggested that m-calpain could be involved in myoblast fusion via desmin cleavage. This hypothesis was confirmed by the results obtained after calpeptin treatment. In the presence of this cell-penetrating inhibitor of calpains, desmin seems not to be degraded. Taking into account the observations obtained after different hydrolysis assays and as compared to those observed on cultured cells, it seems conceivable that m-calpain would be able to initiate desmin cleavage leading to the formation of proteolytic fragments which should be immediately degraded.
Elamrani
N
N
ISTAB, UA-INRA Laboratoire de Biochimie et Technologie des Aliments, Université Bordeaux I, Talence, France.
Brustis
J J
JJ
Dourdin
N
N
Balcerzak
D
D
Poussard
S
S
Cottin
P
P
Ducastaing
A
A
D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002154:Calpain / Q000378:metabolism; D002459:Cell Fusion; D002478:Cells, Cultured; D003893:Desmin / Q000378:metabolism*; D004151:Dipeptides / Q000494:pharmacology; D007382:Intermediate Filaments / Q000378:metabolism; D009124:Muscle Proteins / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000166:cytology* / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar
D003893:Desmin; D004151:Dipeptides; D009124:Muscle Proteins; C071482:calpeptin; D002154:Calpain; D002118:Calcium