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Modulation of differentiation in vitro. II. Influence of cell spreading and surface events on myogenesis

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Summary

We examined the influence of attachment and spreading on myogenesis by adding polylysine-covered beads at different times after plating the cells on a plastic substratum. We show that polylysine per se acting on the cell surface can modulate myogenesis independently of cell spreading. Thus cell shape would not be the limiting factor for the division and differentiation of L6 myoblasts. Multinucleation of the cells was found to be first enhanced by the addition of polylysine-covered beads to replicating myoblasts, although the final percentage of fusion attained by these cultures was lower than in the controls. A similar phenomenon was observed concerning myosin synthesis. No such effect could be observed when the beads were added to a nonfusing mutant or to fibroblasts. Our results show that this phenomenon is specific. We postulate that some of the surface molecules necessary for this process appear on myoblasts shortly before they fuse.

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References

  1. Smith, A. A.; Searls, R. L.; Hilfer, S. R. Differential accumulation of extracellular materials beneath the ectoderm during development of the embryonic chick limb and flank regions. Dev. Biol. 46: 222–226; 1979.

    Article  Google Scholar 

  2. Hay, E. D.; Meier, S. Stimulation of corneal differentiation by interaction between cell surface and extracellular matrix. II. Further studies on the nature of transfilter “induction”. Dev. Biol. 52: 141–157; 1976.

    Article  PubMed  CAS  Google Scholar 

  3. Truppe, W.; Basner, R.; Von Figura, K.; Kresse, H. Uptake of hyaluronate by cultured cells. Biochem. Biophys. Res. Commun. 78: 713–719; 1977.

    PubMed  CAS  Google Scholar 

  4. Huang, D. Effect of extracellular chondroitin surface on cultured chrondrocytes. J. Cell Biol. 62: 881–886; 1974.

    Article  PubMed  CAS  Google Scholar 

  5. Nathanson, M. A.; Hay, E. D. Analysis of carttilage differentiation from skeletal muscle grown on bone matrix. II. Chondroitin sulfate synthesis and reaction to exogenous glycosaminoglycans. Dev. Biol. 78: 332–351; 1980.

    Article  PubMed  CAS  Google Scholar 

  6. Elson, M. F.; Ingwall, S. J. The cell substratum modulates skeletal muscle differentiation. J. Supramol. Struct. 14: 313–328; 1980.

    Article  PubMed  CAS  Google Scholar 

  7. MacBride, R. G.; Przybylski, R. J. Purified lectin from skeletal muscle inhibits myotube formation in vitro. J. Cell Biol. 85: 617–625; 1980.

    Article  PubMed  CAS  Google Scholar 

  8. Chen, L. B.; Murray, A.; Segal, R. A.; Bushnell, A.; Walsh, M. L. Studies on intercellular LETS glycoprotein matrices. Cell 14: 377–391; 1978.

    Article  PubMed  CAS  Google Scholar 

  9. Podleski, T. R.; Greenberg, I.; Schlessinger, J.; Yamada, K. M. Fibronectin delays the fusion of L6 myoblasts. Exp. Cell Res. 122: 317–326; 1979.

    Article  PubMed  CAS  Google Scholar 

  10. Hauschka, S. D.; Konigsberg, I. R. The influence of collagen on the development of muscle clones. Proc. Natl. Acad. Sci. USA 55: 119–126; 1966.

    Article  PubMed  CAS  Google Scholar 

  11. Delain, D.; Wahrmann, J. P.; Gros, F. Influence of external diffusible factors on myogenesis of the cells of the line L6. Exp. Cell Res. 131: 217–224; 1981.

    Article  PubMed  CAS  Google Scholar 

  12. Emerman, J.; Enami, J.; Pitelka, D. R.; Nandi, S. Hormonal effects on intracellular and secreted casein in cultures of mouse mammary epithelial cells on floating collagen membranes. Proc. Natl. Acad. Sci. USA 74:4466–4470; 1977.

    Article  PubMed  CAS  Google Scholar 

  13. macieira-Coelho, A.; Avrameas, S. Modulation of cell behavior in vitro by the substratum in fibroblastic and leukemic mouse cell lines. Proc. Natl. Acad. Sci. USA 69: 2469–2473; 1972.

    Article  PubMed  CAS  Google Scholar 

  14. Folkman, J.; Moscona, A. Role of cell shape in growth control. Nature 273: 345–349; 1978.

    Article  PubMed  CAS  Google Scholar 

  15. Ben-Ze'ev, A.; Farmer, S. R.; Penman, S. Protein synthesis requires cell-surface contact while nuclear events respond to cell shape in anchorage-dependent fibroblasts. Cell 21: 365–372; 1980.

    Article  PubMed  Google Scholar 

  16. Wahrmann, J. P.; Delain, D.; Bournoutian, C.; Macieira-Coelho, A. Modulation of differentiation in vitro. I. Influence of the attachment surface on myogenesis. In Vitro 17: 752–762; 1981.

    Article  PubMed  CAS  Google Scholar 

  17. Somers, D. G.; Pearson, M. L.; Ingles, C. J. Isolation and characterization of an alpha amanitan-resistant rat myoblast mutant cell line possessing alpha amanitin-resistant RNA polymerase. J. Biol. Chem. 250: 4825–4831; 1975.

    PubMed  CAS  Google Scholar 

  18. Kapuscinski, J.; Skoczylas, B. Simple and rapid fluorometric method for DNA microassay. Anal. Biochem. 83: 252–257; 1977.

    Article  PubMed  CAS  Google Scholar 

  19. Emerson, C. P.; Beckner, S. K. Activation of myosin synthesis in fusing and mononucleated myoblasts. J. Mol. Biol. (London) 93: 431–448; 1975.

    CAS  Google Scholar 

  20. Porzio, L. A.; Pearson, A. M. Improved resolution of myofibrilar proteins with sodium dodecyl sulfate polyacrylamide electrophoresis. Biochim. Biophys. Acta 490: 27–37; 1977.

    PubMed  CAS  Google Scholar 

  21. Cohen, A.; Buckingham, M.; Gros, F. A modified assay procedure for revealing the M form of creatine kinase in cultured muscle cells. Exp. Cell Res. 115: 201–206; 1978.

    Article  PubMed  CAS  Google Scholar 

  22. Ternynck, T.; Avrameas, S. Polymerization and immobilization of proteins using ethylchloroformate and glutaraldehyde. Scand. J. Immunol. 3: 29–35; 1976.

    Article  CAS  Google Scholar 

  23. Morris, G. E.; Cooke, A.; Cole, R. J. Isoenzymes of creatine phosphokinase activity during myogenesis in vitro. Exp. Cell Res. 74: 582–584; 1972.

    Article  PubMed  CAS  Google Scholar 

  24. Turner, D. C.; Maier, V.; Eppenberger, H. M. Creatine kinase and aldolase isoenzymes transitions in cultures of chick skeletal muscle cells. Dev. Biol. 37: 63–90; 1974.

    Article  PubMed  CAS  Google Scholar 

  25. Luzzati, D.; Loomis, W. F.; Drugeon, G.; Wahrmann, J. P. Synthèse des protéines caractéristiques de la cellule musculaire au cours de la différenciation d'une lignée établie de cellules myogénique et de quelques mutants conditionels de différenciation. In: Schapira, G., Dreyfus, J. C., Kruh, J., eds. Normal and pathological protein synthesis in higher organisms. Paris: INSERM; 1973: 475–485.

    Google Scholar 

  26. Wahrmann, J. P.; Sénéchal, H.; Etienne-Decerf, J., Winand, R. J. Changes in cell surface glycopeptides during myogenesis in wild type and nonfusing mutant L6 rat myoblasts. FEBS Lett. 115: 230–234; 1980.

    Article  PubMed  CAS  Google Scholar 

  27. Sénéchal, H.; Schapira, G.; Wahrmann, J. P. Changes in plasma membrane glycoproteins during differentiation of an established myoblast cell line and a nonfusing α-amanitin resistant mutant. Exp. Cell Res. 138: 355–365; 1982.

    Article  PubMed  Google Scholar 

  28. Sénéchal, H.; Delain, D.; Schapira, G.; Wahrmann, J. P. Alterations on glycosylation of plasma membrane proteins during myogenesis. Exp. Cell Res. 147: 341–350; 1983.

    Article  PubMed  Google Scholar 

  29. Macieira-Coelho, A.; Berumen, L.; Avrameas, S. Properties of protein polymers as substratum for cell growth in vitro. J. Cell. Physiol. 83: 379–388; 1974.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Institut de Pathologie Moléculaire, (INSERM U 15 & LA CNRS), 75014, Paris

    H. Sénéchal, J. P. Wahrmann & D. Delain

  2. Laboratoire de Pathologie Cellulaire, (I.C.I.G.), 94804, Villejuif, France

    A. Macieira-Coelho

Authors
  1. H. Sénéchal
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  2. J. P. Wahrmann
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  3. D. Delain
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  4. A. Macieira-Coelho
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Additional information

This work was supported by the American Dystrophy Association and by the Association pour la Recherche sur le Cancer (ARC) (Contract no 3050).

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Sénéchal, H., Wahrmann, J.P., Delain, D. et al. Modulation of differentiation in vitro. II. Influence of cell spreading and surface events on myogenesis. In Vitro 20, 692–698 (1984). https://doi.org/10.1007/BF02618874

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  • DOI: https://doi.org/10.1007/BF02618874

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