Potential m-calpain substrates during myoblast fusion

N Dourdin, D Balcerzak, J J Brustis, S Poussard, P Cottin, A Ducastaing

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Abstract

Many studies have demonstrated that m-calpain was implicated in cell membrane reorganization-related phenomena during fusion via a regulation by calpastatin, the specific Ca2+-dependent proteolytic inhibitor. However, the real biological role of this protease is unclear because many targeted proteins are still unknown. Using different digestion experiments we have demonstrated that desmin, vimentin, talin, and fibronectin represent very good substrates for this proteinase capable of cleaving them in fragments which are immediately degraded by other enzymatic systems. Concerning intermediate filaments, we showed that during the phenomenon of fusion, the amount of desmin was significantly reduced while the concentration of vimentin presented a steady level. On the other hand, we have conducted biological assays on cultured myoblasts supplemented by exogenous factors such as calpain inhibitors or antisense oligonucleotides capable of stimulating or inhibiting m-calpain activity. The effect of such factors on fusion and concomitantly on the targeted substrates was analyzed and quantified. When m-calpain activity and myoblast fusion were prevented by addition of calpain inhibitors entering the cells, the amounts of desmin, talin, and fibronectin were increased, whereas the amount of vimentin was unchanged. Using antisense strategy, similar results were obtained. In addition, when the phenomenon of fusion was enhanced by preventing calpastatin synthesis, the amounts of desmin, talin, and fibronectin were significantly reduced. Taken together, these results support the hypothesis that m-calpain is involved in myoblast fusion by cleaving certain proteins identified here. This cleavage could modify membrane and cytoskeleton organization for the myoblasts to fuse, (C) 1999 Academic Press.

Original languageEnglish
Pages (from-to)433-442
Number of pages10
JournalExperimental Cell Research
Volume246
Issue number2
Publication statusPublished - 1 Feb 1999

Keywords

  • m-calpain
  • calpain inhibitors
  • desmin
  • fibronectin
  • talin
  • vimentin
  • antisense oligodeoxyribonucleotides
  • myoblast culture
  • fusion
  • exteriorized m-calpain
  • intermediate filaments
  • muscle-cells
  • fibronectin degradation
  • expression
  • protease
  • cleavage
  • proteins

Cite this

Dourdin, N., Balcerzak, D., Brustis, J. J., Poussard, S., Cottin, P., & Ducastaing, A. (1999). Potential m-calpain substrates during myoblast fusion. Experimental Cell Research, 246(2), 433-442.