Skip to main content
Log in

Changes in myosin distribution in dedifferentiating and redifferentiating smooth muscle cells in tissue culture

  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Summary

Isolated smooth muscle cells and fibroblasts from the newborn guinea-pig vas deferens were grown in culture. In the first 2 days, all cells characterized as smooth muscle by phase-contrast microscopy reacted intensely with fluoresceinated antibodies against smooth muscle myosin. The fluorescence was in the form of particles (termed here “myosin aggregates”), which were often aligned to give the cell a striated appearance. After 3–5 days, coarse fluorescent fibrils were also visible. These were termed “attachment fibrils” (“A-fibrils”) since they were thought to represent myosin in microfilament bundles. Between 6 and 7 days in culture, the smooth muscle cells began to dedifferentiate morphologically. At this time, the “myosin aggregates” became clumped and less intensely fluorescent. “A-fibrils” also decreased in fluorescence intensity. By 8 days in culture, the dedifferentiated cells had undergone intense proliferation and gave only a minimal reaction with myosin antibodies. However, when a confluent monolayer of cells formed on day 9 or 10, they immediately began to redifferentiate ultrastructurally and to regain immunofluorescence in both “myosin aggregates” and “A-fibrils”. Throughout the entire culture period, cells characterized as fibroblasts by phase contrast microscopy gave only a weak reaction with fluoresceinated antibodies to myosin showing “A-fibrils” but no “myosin aggregates”.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Campbell, G.R., Uehara, Y., Mark, G., Burnstock, G.: Fine structure of smooth muscle cells grown in tissue culture. J. Cell Biol. 49, 21–34 (1971)

    Google Scholar 

  • Chamley, J.H., Campbell, G.R.: Isolated ureteral smooth muscle cells in culture. Including their interaction with intrinsic and extrinsic nerves. Cytobiol. (in press) (1975)

  • Chamley, J.H., Campbell, G.R., Burnstock, G.: Dedifferentiation, redifferentiation and bundle formation of smooth muscle cells in tissue culture: the influence of cell number and nerve fibres. J. Embryol. exp. Morph. 32, 297–323 (1974)

    Google Scholar 

  • Fisher-Dzoga, K., Jones, R.M., Vesselinovitch, D., Wissler, R.W.: Ultrastructural and immunohistochemical studies of primary cultures of aortic medial cells. Exp. molec. Path. 18, 162–176 (1973)

    Google Scholar 

  • Goldman, R.D., Lazarides, E., Pollack, R., Weber, K.: The distribution of actin in non-muscle cells. Exp. Cell Res. 90, 333–344 (1975)

    Google Scholar 

  • Gröschel-Stewart, U., Chamley, J.H., McConnell, J.D., Burnstock, G.: Comparison of the reaction of cultured smooth and cardiac muscle cells and fibroblasts to specific antibodies to myosin. Histochemistry 43, 215–224 (1975a)

    Google Scholar 

  • Gröschel-Stewart, U., Gröschel, D.: Immunological evidence for the presence of smooth muscle-type contractile fibres in mouse macrophages. Experientia (Basel) 30, 1152–1153 (1974)

    Google Scholar 

  • Gröschel-Stewart, U., Schreiber, J., Mahlmeister, Chr., Weber, K.: Production of specific antibodies to contractile proteins, and their use in immunofluorescence microscopy. I. Antibodies to smooth and striated chicken muscle myosin. Histochemistry (in press) (1975b)

  • Hofmann, W., Goger, D.: Report on the differentiation of vascular wall smooth muscle cells with the aid of immunofluorescence. Virchows Arch. A. Pathol. Anat. and Histol. 363, 225–231 (1974)

    Google Scholar 

  • Lazarides, E.: Tropomyosin antibody: The specific localization of tropomyosin in nonmuscle cells. J. Cell Biol. 65, 549–561 (1975)

    Google Scholar 

  • Lazarides, E., Weber, K.: Actin antibody: The specific visualization of actin filaments in non-muscle cells. Proc. nat. Acad. Sci. (Wash.) 71, 2268–2272 (1974)

    Google Scholar 

  • Mark, G.E., Chamley, J.H., Burnstock, G.: Interactions between autonomic nerves and smooth and cardiac muscle cells in tissue culture. Develop. Biol. 32, 194–200 (1973)

    Google Scholar 

  • Painter, R.G., Sheetz, M., Singer, S.J.: Detection and ultrastructural localization of human smooth muscle myosin-like molecules in human non-muscle cells by specific antibodies. Proc. nat. Acad. Sci. (Wash.) 72, 1359–1363 (1975)

    Google Scholar 

  • Pollack, R., Osborn, M., Weber, K.: Patterns of organization of actin and myosin in normal and transformed cultured cells. Proc. nat. Acad. Sci. (Wash.) 72, 994–998 (1975)

    Google Scholar 

  • Small, J.V.: Contractile units in vertebrate smooth muscle cells. Nature (Lond.) 249, 324–327 (1974)

    Google Scholar 

  • Weber, K., Gröschel-Stewart, U.: Antibody to myosin. The specific visualization of myosin-containing filaments in non-muscle cells. Proc. nat. Acad. Sci. (Wash.) 71, 4561–4564 (1974)

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

The authors wish to thank Miss J.D. McConnell for excellent technical assistance.

Supported by the Deutsche Forschungsgemeinschaft.

Supported by The Life Insurance Medical Research Fund of Australia and New Zealand.

Supported by the National Heart Foundation of Australia.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gröschel-Stewart, U., Chamley, J.H., Campbell, G.R. et al. Changes in myosin distribution in dedifferentiating and redifferentiating smooth muscle cells in tissue culture. Cell Tissue Res. 165, 13–22 (1975). https://doi.org/10.1007/BF00222796

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00222796

Key words

Navigation