Calpain and myogenesis: development of a convenient cell culture model.
Fiche publication
Date publication
mai 2002
Journal
Biology of the cell
Auteurs
Membres identifiés du Cancéropôle Est :
Pr DEDIEU Stéphane
Tous les auteurs :
Dedieu S, Dourdin N, Dargelos E, Poussard S, Veschambre P, Cottin P, Brustis JJ
Lien Pubmed
Résumé
Previous studies have led us to hypothesize that m-calpain plays a pivotal role in myoblast fusion through its involvement in cell membrane and cytoskeleton component reorganization. To support this hypothesis, a convenient and simple myoblast culture model using frozen embryonic myoblasts was developed, which resolved a number of problems inherent to cell primary culture. Biological assays on cultured myoblasts using different media to define the characteristics of the fusion process were first conducted. Proteinase was detectable before the initiation of the fusion process and was closely correlated to the phenomenon of fusion under each culture condition studied. In addition, the study of calpastatin showed that the initiation of fusion does not require a decrease in the level of this endogenous inhibitor of calpains and also confirmed that calpastatin may be implicated in the determination of the end of fusion. On the other hand, analysis of the evolution of myogenic factors revealed that myogenins, MyoD and Myf5, increase very significantly during the formation of multinucleated myotubes. Moreover, the antisense technique against myogenin is capable of preventing the process of fusion by 50%, confirming the pivotal role of this factor in the early stages of differentiation. The possible role of myogenic regulator factors on m-calpain gene expression is discussed.
Mots clés
Animals, Calcium-Binding Proteins, metabolism, Calpain, physiology, Cell Fusion, Cells, Cultured, Models, Biological, Muscle Development, physiology, Myoblasts, physiology, Myogenin, antagonists & inhibitors, Rats, Rats, Wistar
Référence
Biol. Cell. 2002 May;94(2):65-76