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Calpain and myogenesis: development of a convenient cell culture model
Author(s) -
Dedieu Stéphane,
Dourdin Nathalie,
Dargelos Elise,
Poussard Sylvie,
Veschambre Philippe,
Cottin Patrick,
Brustis JeanJacques
Publication year - 2002
Publication title -
biology of the cell
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 85
eISSN - 1768-322X
pISSN - 0248-4900
DOI - 10.1016/s0248-4900(02)01183-8
Subject(s) - myogenesis , calpastatin , myf5 , myogenin , calpain , cell fusion , biology , myod , myocyte , microbiology and biotechnology , multinucleate , cell culture , fusion protein , myod protein , genetics , biochemistry , gene , recombinant dna , enzyme
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.