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. 1988 Aug 1;107(2):761–769. doi: 10.1083/jcb.107.2.761

Cell surface fibroblast growth factor and epidermal growth factor receptors are permanently lost during skeletal muscle terminal differentiation in culture

PMCID: PMC2115215  PMID: 2843547

Abstract

One characteristic of skeletal muscle differentiation is the conversion of proliferating cells to a population that is irreversibly postmitotic. This developmental change can be induced in vitro by depriving the cultures of specific mitogens such as fibroblast growth factor (FGF). Analysis of cell surface FGF receptor (FGFR) in several adult mouse muscle cell lines and epidermal growth factor receptor (EGFR) in mouse MM14 cells reveals a correlation between receptor loss and the acquisition of a postmitotic phenotype. Quiescent MM14 cells, mitogen-depleted, differentiation-defective MM14 cells, and differentiated BC3H1 muscle cells (a line that fails to become postmitotic upon differentiation) retained their cell surface FGFR. These results indicate that FGFR loss is not associated with either reversible cessation of muscle cell proliferation or biochemical differentiation and thus, further support a correlation between receptor loss and acquisition of a postmitotic phenotype. Comparison of the kinetics for growth factor receptor loss and for commitment of MM14 cells to a postmitotic phenotype reveals that FGFR rises transiently from approximately 700 receptors/cell to a maximum of approximately 2,000 receptors/cell 12 h after FGF removal, when at the same time, greater than 95% of the cells are postmitotic. FGFR levels then decline to undetectable levels by 24 h after FGF removal. During the interval in which FGFR increases and then disappears there is no change in its affinity for FGF. The transient increase in growth factor receptors appears to be due to a decrease in ligand-mediated internalization because EGFR, which undergoes an immediate decline when cultures are deprived of FGF (Lim, R. W., and S. D. Hauschka. 1984. J. Cell Biol. 98:739-747), exhibits a similar transient rise when cultures are grown in media containing both EGF and FGF before switching the cells to media without these added factors. These results indicate that the loss of certain growth factor receptors is a specific phenotype acquired during skeletal muscle differentiation, but they do not resolve whether regulation of FGFR number is causal for initiation of the postmitotic phenotype. A general model is presented in the discussion.

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Selected References

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  1. Aharonov A., Pruss R. M., Herschman H. R. Epidermal growth factor. Relationship between receptor regulation and mitogenesis in 3T3 cells. J Biol Chem. 1978 Jun 10;253(11):3970–3977. [PubMed] [Google Scholar]
  2. Bader D., Masaki T., Fischman D. A. Immunochemical analysis of myosin heavy chain during avian myogenesis in vivo and in vitro. J Cell Biol. 1982 Dec;95(3):763–770. doi: 10.1083/jcb.95.3.763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baird A., Ling N. Fibroblast growth factors are present in the extracellular matrix produced by endothelial cells in vitro: implications for a role of heparinase-like enzymes in the neovascular response. Biochem Biophys Res Commun. 1987 Jan 30;142(2):428–435. doi: 10.1016/0006-291x(87)90292-0. [DOI] [PubMed] [Google Scholar]
  4. Beguinot F., Kahn C. R., Moses A. C., Smith R. J. Distinct biologically active receptors for insulin, insulin-like growth factor I, and insulin-like growth factor II in cultured skeletal muscle cells. J Biol Chem. 1985 Dec 15;260(29):15892–15898. [PubMed] [Google Scholar]
  5. Bischoff R. A satellite cell mitogen from crushed adult muscle. Dev Biol. 1986 May;115(1):140–147. doi: 10.1016/0012-1606(86)90235-6. [DOI] [PubMed] [Google Scholar]
  6. Bischoff R. Proliferation of muscle satellite cells on intact myofibers in culture. Dev Biol. 1986 May;115(1):129–139. doi: 10.1016/0012-1606(86)90234-4. [DOI] [PubMed] [Google Scholar]
  7. Bischoff R. Regeneration of single skeletal muscle fibers in vitro. Anat Rec. 1975 Jun;182(2):215–235. doi: 10.1002/ar.1091820207. [DOI] [PubMed] [Google Scholar]
  8. Blau H. M., Pavlath G. K., Hardeman E. C., Chiu C. P., Silberstein L., Webster S. G., Miller S. C., Webster C. Plasticity of the differentiated state. Science. 1985 Nov 15;230(4727):758–766. doi: 10.1126/science.2414846. [DOI] [PubMed] [Google Scholar]
  9. Carpenter G., Cohen S. Human epidermal growth factor and the proliferation of human fibroblasts. J Cell Physiol. 1976 Jun;88(2):227–237. doi: 10.1002/jcp.1040880212. [DOI] [PubMed] [Google Scholar]
  10. Clegg C. H., Hauschka S. D. Heterokaryon analysis of muscle differentiation: regulation of the postmitotic state. J Cell Biol. 1987 Aug;105(2):937–947. doi: 10.1083/jcb.105.2.937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Clegg C. H., Linkhart T. A., Olwin B. B., Hauschka S. D. Growth factor control of skeletal muscle differentiation: commitment to terminal differentiation occurs in G1 phase and is repressed by fibroblast growth factor. J Cell Biol. 1987 Aug;105(2):949–956. doi: 10.1083/jcb.105.2.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Endo T., Nadal-Ginard B. Transcriptional and posttranscriptional control of c-myc during myogenesis: its mRNA remains inducible in differentiated cells and does not suppress the differentiated phenotype. Mol Cell Biol. 1986 May;6(5):1412–1421. doi: 10.1128/mcb.6.5.1412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ewton D. Z., Spizz G., Olson E. N., Florini J. R. Decrease in transforming growth factor-beta binding and action during differentiation in muscle cells. J Biol Chem. 1988 Mar 15;263(8):4029–4032. [PubMed] [Google Scholar]
  14. Florini J. R., Roberts A. B., Ewton D. Z., Falen S. L., Flanders K. C., Sporn M. B. Transforming growth factor-beta. A very potent inhibitor of myoblast differentiation, identical to the differentiation inhibitor secreted by Buffalo rat liver cells. J Biol Chem. 1986 Dec 15;261(35):16509–16513. [PubMed] [Google Scholar]
  15. Gospodarowicz D., Weseman J., Moran J. S., Lindstrom J. Effect of fibroblast growth factor on the division and fusion of bovine myoblasts. J Cell Biol. 1976 Aug;70(2 Pt 1):395–405. doi: 10.1083/jcb.70.2.395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kardami E., Spector D., Strohman R. C. Myogenic growth factor present in skeletal muscle is purified by heparin-affinity chromatography. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8044–8047. doi: 10.1073/pnas.82.23.8044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kardami E., Spector D., Strohman R. C. Selected muscle and nerve extracts contain an activity which stimulates myoblast proliferation and which is distinct from transferrin. Dev Biol. 1985 Dec;112(2):353–358. doi: 10.1016/0012-1606(85)90406-3. [DOI] [PubMed] [Google Scholar]
  18. Konigsberg I. R. Diffusion-mediated control of myoblast fusion. Dev Biol. 1971 Sep;26(1):133–152. doi: 10.1016/0012-1606(71)90113-8. [DOI] [PubMed] [Google Scholar]
  19. Konigsberg U. R., Lipton B. H., Konigsberg I. R. The regenerative response of single mature muscle fibers isolated in vitro. Dev Biol. 1975 Aug;45(2):260–275. doi: 10.1016/0012-1606(75)90065-2. [DOI] [PubMed] [Google Scholar]
  20. Lathrop B., Olson E., Glaser L. Control by fibroblast growth factor of differentiation in the BC3H1 muscle cell line. J Cell Biol. 1985 May;100(5):1540–1547. doi: 10.1083/jcb.100.5.1540. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lathrop B., Thomas K., Glaser L. Control of myogenic differentiation by fibroblast growth factor is mediated by position in the G1 phase of the cell cycle. J Cell Biol. 1985 Dec;101(6):2194–2198. doi: 10.1083/jcb.101.6.2194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lim R. W., Hauschka S. D. A rapid decrease in epidermal growth factor-binding capacity accompanies the terminal differentiation of mouse myoblasts in vitro. J Cell Biol. 1984 Feb;98(2):739–747. doi: 10.1083/jcb.98.2.739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lim R. W., Hauschka S. D. EGF responsiveness and receptor regulation in normal and differentiation-defective mouse myoblasts. Dev Biol. 1984 Sep;105(1):48–58. doi: 10.1016/0012-1606(84)90260-4. [DOI] [PubMed] [Google Scholar]
  24. Linkhart T. A., Clegg C. H., Hauschika S. D. Myogenic differentiation in permanent clonal mouse myoblast cell lines: regulation by macromolecular growth factors in the culture medium. Dev Biol. 1981 Aug;86(1):19–30. doi: 10.1016/0012-1606(81)90311-0. [DOI] [PubMed] [Google Scholar]
  25. Linkhart T. A., Clegg C. H., Hauschka S. D. Control of mouse myoblast commitment to terminal differentiation by mitogens. J Supramol Struct. 1980;14(4):483–498. doi: 10.1002/jss.400140407. [DOI] [PubMed] [Google Scholar]
  26. MAURO A. Satellite cell of skeletal muscle fibers. J Biophys Biochem Cytol. 1961 Feb;9:493–495. doi: 10.1083/jcb.9.2.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Massagué J., Cheifetz S., Endo T., Nadal-Ginard B. Type beta transforming growth factor is an inhibitor of myogenic differentiation. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8206–8210. doi: 10.1073/pnas.83.21.8206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nadal-Ginard B. Commitment, fusion and biochemical differentiation of a myogenic cell line in the absence of DNA synthesis. Cell. 1978 Nov;15(3):855–864. doi: 10.1016/0092-8674(78)90270-2. [DOI] [PubMed] [Google Scholar]
  29. Olson E. N., Sternberg E., Hu J. S., Spizz G., Wilcox C. Regulation of myogenic differentiation by type beta transforming growth factor. J Cell Biol. 1986 Nov;103(5):1799–1805. doi: 10.1083/jcb.103.5.1799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Olwin B. B., Hauschka S. D. Identification of the fibroblast growth factor receptor of Swiss 3T3 cells and mouse skeletal muscle myoblasts. Biochemistry. 1986 Jun 17;25(12):3487–3492. doi: 10.1021/bi00360a001. [DOI] [PubMed] [Google Scholar]
  31. Schubert D., Harris A. J., Devine C. E., Heinemann S. Characterization of a unique muscle cell line. J Cell Biol. 1974 May;61(2):398–413. doi: 10.1083/jcb.61.2.398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schultz E. Fine structure of satellite cells in growing skeletal muscle. Am J Anat. 1976 Sep;147(1):49–70. doi: 10.1002/aja.1001470105. [DOI] [PubMed] [Google Scholar]
  33. Shechter Y., Hernaez L., Cuatrecasas P. Epidermal growth factor: biological activity requires persistent occupation of high-affinity cell surface receptors. Proc Natl Acad Sci U S A. 1978 Dec;75(12):5788–5791. doi: 10.1073/pnas.75.12.5788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Snow M. H. An autoradiographic study of satellite cell differentiation into regenerating myotubes following transplantation of muscles in young rats. Cell Tissue Res. 1978 Jan 31;186(3):535–540. doi: 10.1007/BF00224941. [DOI] [PubMed] [Google Scholar]
  35. Vlodavsky I., Folkman J., Sullivan R., Fridman R., Ishai-Michaeli R., Sasse J., Klagsbrun M. Endothelial cell-derived basic fibroblast growth factor: synthesis and deposition into subendothelial extracellular matrix. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2292–2296. doi: 10.1073/pnas.84.8.2292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yaffe D. Developmental changes preceding cell fusion during muscle differentiation in vitro. Exp Cell Res. 1971 May;66(1):33–48. doi: 10.1016/s0014-4827(71)80008-3. [DOI] [PubMed] [Google Scholar]
  37. Yaffe D., Saxel O. Serial passaging and differentiation of myogenic cells isolated from dystrophic mouse muscle. Nature. 1977 Dec 22;270(5639):725–727. doi: 10.1038/270725a0. [DOI] [PubMed] [Google Scholar]

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