Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1984 Feb 1;98(2):739–747. doi: 10.1083/jcb.98.2.739

A rapid decrease in epidermal growth factor-binding capacity accompanies the terminal differentiation of mouse myoblasts in vitro

PMCID: PMC2113105  PMID: 6319433

Abstract

Specific mitogens stimulate the proliferation and repress the differentiation of mouse myoblasts (MM14). When mitogens are depleted, MM14 cells cease proliferation, commit to terminal differentiation, and become refractory to growth stimulation. The behavior of mitogen receptors during the transition from a proliferative to a permanently postmitotic state was examined using the epidermal growth factor receptor (EGFR) as a model system. Whereas proliferating myoblasts bound substantial amounts of EGF, their binding capacity declined rapidly upon exposure to low-mitogen medium. The decline became irreversible when a cell differentiated. Within 24 h, less than 5% of the original EGF binding capacity remained. Since the ability to internalize and degrade bound EGF was unaffected, the change presumably reflected a decrease in EGFR availability. Several observations indicated that loss of EGFR following mitogen removal is related to differentiation rather than the result of starvation or cell-cycle arrest. First, the decline is correlated with the absence of a single mitogen (fibroblast growth factor) and is independent of serum concentrations. Second, myoblasts that are either cycling through G1 or arrested at G0, but prevented from differentiating, all bind large amounts of EGF. These findings suggest that specific reduction in mitogen receptors could be part of a mechanism whereby terminally differentiating cells become refractory to mitogenic stimulation.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  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. Carpenter G., Cohen S. 125I-labeled human epidermal growth factor. Binding, internalization, and degradation in human fibroblasts. J Cell Biol. 1976 Oct;71(1):159–171. doi: 10.1083/jcb.71.1.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. De Larco J. E., Todaro G. J. Sarcoma growth factor (SGF): specific binding to epidermal growth factor (EGF) membrane receptors. J Cell Physiol. 1980 Feb;102(2):267–277. doi: 10.1002/jcp.1041020218. [DOI] [PubMed] [Google Scholar]
  4. Devlin B. H., Konigsberg I. R. Reentry into the cell cycle of differentiated skeletal myocytes. Dev Biol. 1983 Jan;95(1):175–192. doi: 10.1016/0012-1606(83)90016-7. [DOI] [PubMed] [Google Scholar]
  5. Ewton D. Z., Florini J. R. Relative effects of the somatomedins, multiplication-stimulating activity, and growth hormone on myoblasts and myotubes in culture. Endocrinology. 1980 Feb;106(2):577–583. doi: 10.1210/endo-106-2-577. [DOI] [PubMed] [Google Scholar]
  6. Grove B. K., Schwartz G., Stockdale F. E. Quantitation of changes in cell surface determinants during skeletal muscle cell differentiation using monospecific antibody. J Supramol Struct Cell Biochem. 1981;17(2):147–152. doi: 10.1002/jsscb.380170205. [DOI] [PubMed] [Google Scholar]
  7. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  8. Haigler H. T., Maxfield F. R., Willingham M. C., Pastan I. Dansylcadaverine inhibits internalization of 125I-epidermal growth factor in BALB 3T3 cells. J Biol Chem. 1980 Feb 25;255(4):1239–1241. [PubMed] [Google Scholar]
  9. Hollenberg M. D., Cuatrecasas P. Insulin and epidermal growth factor. Human fibroblast receptors related to deoxyribonucleic acid synthesis and amino acid uptake. J Biol Chem. 1975 May 25;250(10):3845–3853. [PubMed] [Google Scholar]
  10. Huff K., End D., Guroff G. Nerve growth factor-induced alteration in the response of PC12 pheochromocytoma cells to epidermal growth factor. J Cell Biol. 1981 Jan;88(1):189–198. doi: 10.1083/jcb.88.1.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Johnson L. K., Baxter J. D., Vlodavsky I., Gospodarowicz D. Epidermal growth factor and expression of specific genes: effects on cultured rat pituitary cells are dissociable from the mitogenic response. Proc Natl Acad Sci U S A. 1980 Jan;77(1):394–398. doi: 10.1073/pnas.77.1.394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. King A. C., Hernaez-Davis L., Cuatrecasas P. Lysomotropic amines cause intracellular accumulation of receptors for epidermal growth factor. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3283–3287. doi: 10.1073/pnas.77.6.3283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Konigsberg I. R., Sollmann P. A., Mixter L. O. The duration of the terminal G1 of fusing myoblasts. Dev Biol. 1978 Mar;63(1):11–26. doi: 10.1016/0012-1606(78)90109-4. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Lloyd C. E., Ascoli M. On the mechanisms involved in the regulation of the cell-surface receptors for human choriogonadotropin and mouse epidermal growth factor in cultured Leydig tumor cells. J Cell Biol. 1983 Feb;96(2):521–526. doi: 10.1083/jcb.96.2.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Moss M., Norris J. S., Peck E. J., Jr, Schwartz R. J. Alterations in iodinated cell surface proteins during myogenesis. Exp Cell Res. 1978 May;113(2):445–450. doi: 10.1016/0014-4827(78)90388-9. [DOI] [PubMed] [Google Scholar]
  19. Murdoch G. H., Potter E., Nicolaisen A. K., Evans R. M., Rosenfeld M. G. Epidermal growth factor rapidly stimulates prolactin gene transcription. Nature. 1982 Nov 11;300(5888):192–194. doi: 10.1038/300192a0. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. O'Neill M. C., Stockdale F. E. A kinetic analysis of myogenesis in vitro. J Cell Biol. 1972 Jan;52(1):52–65. doi: 10.1083/jcb.52.1.52. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pauw P. G., David J. D. Alterations in surface proteins during myogenesis of a rat myoblast cell line. Dev Biol. 1979 May;70(1):27–38. doi: 10.1016/0012-1606(79)90004-6. [DOI] [PubMed] [Google Scholar]
  23. Rees A. R., Adamson E. D., Graham C. F. Epidermal growth factor receptors increase during the differentiation of embryonal carcinoma cells. Nature. 1979 Sep 27;281(5729):309–311. doi: 10.1038/281309a0. [DOI] [PubMed] [Google Scholar]
  24. Robinson R. A., Volkenant M. E., Ryan R. J., Moses H. L. Decreased epidermal growth factor binding in cells growth arrested in G1 by nutrient deficiency. J Cell Physiol. 1981 Dec;109(3):517–524. doi: 10.1002/jcp.1041090318. [DOI] [PubMed] [Google Scholar]
  25. Rutz R., Haney C., Hauschka S. Spatial analysis of limb bud myogenesis: a proximodistal gradient of muscle colony-forming cells in chick embryo leg buds. Dev Biol. 1982 Apr;90(2):399–411. doi: 10.1016/0012-1606(82)90389-x. [DOI] [PubMed] [Google Scholar]
  26. Sandra A., Przybylski R. J. Ontogeny of insulin binding during chick skeletal myogenesis in vitro. Dev Biol. 1979 Feb;68(2):546–556. doi: 10.1016/0012-1606(79)90225-2. [DOI] [PubMed] [Google Scholar]
  27. Schonberg M., Bilezikian J. P., Apfelbaum M., Benn R. C. Beta-adrenergic receptors and myogenesis. J Cyclic Nucleotide Res. 1978 Feb;4(1):55–69. [PubMed] [Google Scholar]
  28. Walsh F. S., Phillips E. Specific changes in cellular glycoproteins and surface proteins during myogenesis in clonal muscle cells. Dev Biol. 1981 Jan 30;81(2):229–237. doi: 10.1016/0012-1606(81)90286-4. [DOI] [PubMed] [Google Scholar]
  29. White N. K., Bonner P. H., Nelson D. R., Hauschka S. D. Clonal analysis of vertebrate myogenesis. IV. Medium-dependent classification of colony-forming cells. Dev Biol. 1975 Jun;44(2):346–361. doi: 10.1016/0012-1606(75)90405-4. [DOI] [PubMed] [Google Scholar]
  30. Wiley H. S., Cunningham D. D. A steady state model for analyzing the cellular binding, internalization and degradation of polypeptide ligands. Cell. 1981 Aug;25(2):433–440. doi: 10.1016/0092-8674(81)90061-1. [DOI] [PubMed] [Google Scholar]
  31. 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]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES