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FITC-labelled antibody staining of tropomyosin-containing fibrils in smooth, cardiac and skeletal muscle cells, prefusion myoblasts, fibroblasts, endothelial cells and 3T3 cells in culture

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Summary

FITC-labelled antibodies to purified chicken gizzard smooth muscle tropomyosin were prepared and used to stain muscle and non-muscle cells in culture.

Skeletal muscle myoblasts stained both diffusely throughout the cytoplasm and in fine filamentous structures. Once myotubes developed the staining was localized exclusively in the I-band region of the myofibrils. Similarly, cardiac muscle cells stained in the I-band alone.

Primary and subcultured smooth muscle cells, irrespective of their state of differentiation, stained exclusively in long, straight fibrils. The staining of the fibrils was interrupted with stained regions 1–2 μm long and unstained spacings 0.5 μm.

Interrupted fibrils were also observed in fibroblasts and endothelial cells, however their staining reaction was very weak (almost indistinguishable from that with pre-immune serum) and they were few in number.

3T3 cells demonstrated moderate staining in interrupted fibrils. Sheaths of very fine fibrils staining with a similar intensity were also found throughout the cytoplasm. Interruptions in these fine fibrils were often aligned to give the whole cell a striated appearance. Sheaths of fibrils were not found in the other cell types studied

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Authors and Affiliations

  1. Department of Anatomy and Embryology, University College London, London, UK

    Julie Chamley-Campbell, Gordon R. Campbell & Geoffrey Burnstock

  2. Zoologisches Institut, Darmstadt, Bundesrepublik Deutschland

    Ute Gröschel-Stewart

Authors
  1. Julie Chamley-Campbell
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  2. Gordon R. Campbell
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  3. Ute Gröschel-Stewart
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  4. Geoffrey Burnstock
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Additional information

J.C-C. holds a “John Halliday Travelling Fellowship” with the Life Insurance Medical Research Fund of Australia and New Zealand; G.R.C. holds and Overseas research Fellowship with the National Heart Foundation of Australia; U.G.-S. and G.B. are supported by the Deutsche Forschungsgemein-schaft and the Wellcome Trust (London) respectively. We wish to thank Janet D. McConnell and Christine Mahlmeister for excellent technical assistance

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Chamley-Campbell, J., Campbell, G.R., Gröschel-Stewart, U. et al. FITC-labelled antibody staining of tropomyosin-containing fibrils in smooth, cardiac and skeletal muscle cells, prefusion myoblasts, fibroblasts, endothelial cells and 3T3 cells in culture. Cell Tissue Res. 183, 153–166 (1977). https://doi.org/10.1007/BF00226616

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  • DOI: https://doi.org/10.1007/BF00226616

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