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Subcellular localization of newly incorporated myosin in rabbit fast skeletal muscle undergoing stimulation-induced type transformation

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Summary

Immunogold labelling was used to study the distribution of newly synthesized slow muscle myosin (SM) at the ultrastructural level as it replaced fast muscle myosin (FM) in rabbit muscles undergoing stimulation-induced type transformation. Control fast muscle was labelled strongly with antibody to FM and control slow muscle with antibody to SM; label was confined to the A-band. Well-defined differences in the distribution of label within the A-band suggested that the monoclonal antibodies used corresponded to epitopes on different parts of the myosin molecule; this was confirmed by Western blots of subfragments prepared from FM and SM. After 4 weeks of continuous stimulation at 10 Hz, fibres of the tibialis anterior muscle reacted with antibodies to both isoforms; after 6 weeks, labelling was obtained only with antibody to SM. After a 7-week period of stimulation and 3 further weeks of recovery, fibres again reacted with both antibodies. In all positively-labelled sections, the distribution of gold particles was characteristic of the antibody and independent of the origin or history of the fibres. This observation supports the conclusion that newly synthesized myosin is capable of being incorporated throughout the length and cross-section of the A-band.

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References

  • Bennett, P., Craig, R., Starr, R. &Offer, G. (1986) The ultrastructural location of C-protein, X-protein and H-protein in rabbit muscle.J. Musc. Res. Cell Motility 7, 550–67.

    Article  Google Scholar 

  • Bouché, M., Goldfine, S. M. &Fischman, D. A. (1988) Posttranslational incorporation of contractile proteins into myofibrils in a cell-free system.J. Cell Biol. 107, 587–96.

    Article  PubMed  Google Scholar 

  • Brown, J. M. C., Henriksson, J. &Salmons, S. (1989) Restoration of fast muscle characteristics following cessation of chronic stimulation: physiological, histochemical and metabolic changes during slow-to-fast transformation.Proc. Roy. Soc. B. 235, 321–46.

    Google Scholar 

  • Brown, J. &Salmons, S. (1981) Percutaneous control of an implantable muscle stimulator via an optical link.J. Biomed. Eng. 3, 206–8.

    PubMed  Google Scholar 

  • Brown, W. E., Salmons, S. &Whalen, R. G. (1983) The sequential replacement of myosin subunit isoforms during muscle type transformation induced by long-term electrical stimulation.J. Biol. Chem. 258, 14686–92.

    PubMed  Google Scholar 

  • Brown, W. E., Salmons, S. &Whalen, R. G. (1985) Mechanisms underlying the asynchronous replacement of myosin light chain isoforms during stimulation-induced fibre-type transformation of skeletal muscle.FEBS Lett. 192, 235–8.

    Article  PubMed  Google Scholar 

  • Brownson, C., Isenberg, H., Brown, W., Salmons, S. &Edwards, Y. (1988) Changes in skeletal muscle gene transcription induced by chronic stimulation.Musc. Nerve 11, 1183–9.

    Article  Google Scholar 

  • Carlemalm, E., Garavito, R. M. &Villiger, W. (1982) Resin development for electron microscopy and an analysis of embedding at low temperature.J. Microsc. 126, 123–43.

    Google Scholar 

  • Eisenberg, B. R., Brown, J. M. C. &Salmons, S. (1984) Restoration of fast muscle characteristics following cessation of chronic stimulation: the ultrastructure of slow-to-fast transformation.Cell & Tiss. Res. 238, 221–30.

    Google Scholar 

  • Eisenberg, B. R. &Salmons, S. (1981) The reorganisation of subcellular structure in muscle undergoing fast-to-slow type transformation: a stereological study.Cell & Tiss. Res. 220, 449–71.

    Google Scholar 

  • Etlinger, J. D., Zak, R., Fischman, D. A. &Rabinowitz, M. (1975) Isolation of newly synthesized myosin filaments from skeletal muscle homogenates and myofibrils.Nature 255, 259–61.

    Article  PubMed  Google Scholar 

  • Higuchi, H., Funatsu, T., Ishijima, A., Okamura, N. &Ishiwata, S. (1986) Accumulated strain mechanism for length determination of thick filaments in skeletal muscle. I. Experimental bases.J. Musc. Res. Cell Motility 7, 491–500.

    Article  Google Scholar 

  • Huxley, H. E. (1963) Electron microscope studies on the organisation of filaments in striated muscle.J. Molec. Biol. 7, 281–308.

    Google Scholar 

  • Margossian, S. S. &Lowey, S. (1982) Preparation of myosin and its subfragments from rabbit skeletal muscle.Meth. Enzym. 85, 55–70.

    PubMed  Google Scholar 

  • Martin, A. F., Rabinowitz, M., Bough, R., Prior, G. &Zak, R. (1977) Measurements of half-life of rat cardiac myosin heavy chain with leucyl-tRNA used as precursor pool.J. Biol Chem. 252, 3422–9.

    PubMed  Google Scholar 

  • Mittal, B., Sanger, J. M. &Sanger, J. W. (1987) Visualization of myosin in living cells.J. Cell Biol. 105, 1753–60.

    Article  PubMed  Google Scholar 

  • Morkin, E. (1970) Postnatal muscle fiber assembly: localisation of newly synthesized myofibrillar proteins.Science 167, 1499–501.

    PubMed  Google Scholar 

  • Obinata, T., Maruyama, K., Sugita, H., Kohama, K. &Ebashi, S. (1981) Dynamic aspects of structural proteins in vertebrate skeletal muscle.Musc. Nerve 4, 456–88.

    Article  Google Scholar 

  • Pierobon-Bormioli, S., Salviati, G. &Betto, R. (1988) Skeletal muscle endosarcomeric immunoultrastructural localization of titin/connectin, nebulin and ‘Band 4’. InSarcomeric and Non-Sarcomeric Muscles: Basic and Applied Research Prospects for the 90's (edited byCarraro, U.), pp. 511–16. Padua: Unipress.

    Google Scholar 

  • Polak, J. M. &Varndell, I. M. (1984)Immunolabelling for Electron Microscopy. Amsterdam: Elsevier Science Publishers.

    Google Scholar 

  • Roth, J. (1983) The colloidal gold marker system for light and electron microscopic cytochemistry. InTechniques in Immunocytochemistry, Vol. 2 (edited byBullock, G. R. &Petrusz, P.), pp. 218–84. London: Academic Press.

    Google Scholar 

  • Rubinstein, N., Mabuchi, K., Pepe, F., Salmons, S., Gergely, J. &Srėter, F. A. (1978) Use of type-specific antimyosins to demonstrate the transformation of individual fibres in chronically stimulated rabbit fast muscles.J. Cell Biol. 79, 252–61.

    Article  PubMed  Google Scholar 

  • Saad, A. D., Pardee, J. D. &Fischman, D. A. (1986) Dynamic exchange of myosin molecules between thick filaments.Proc. Nat. Acad. Sci. USA 83, 9483–7.

    PubMed  Google Scholar 

  • Salmons, S. (1987) Biochemical and morphological evidence of increased protein and nucleic acid synthesis in chronically stimulated rabbit skeletal muscle.J. Anal. 152, 229.

    Google Scholar 

  • Silver, G. &Etlinger, J. D. (1985) Regulation of myofibrillar accumulation in chick muscle cultures: evidence for the involvement of calcium and lysosomes in non-uniform turnover of contractile proteins.J. Cell Biol. 101, 2383–91.

    Article  PubMed  Google Scholar 

  • Taylor, L. D. &Bandman, E. (1989) Distribution of fast myosin heavy chain isoforms in thick filaments of developing chicken pectoral muscle.J. Cell Biol. (in press).

  • Weeds, A. G. &Taylor, R. S. (1975) Separation of subfragment-1 isoenzymes from rabbit skeletal muscle myosin.Nature 257, 54–6.

    Article  PubMed  Google Scholar 

  • Wenderoth, M. P. &Eisenberg, B. R. (1987) Incorporation of nascent myosin heavy chains into thick filaments of cardiac myocytes in thyroid-treated rabbits.J. Cell Biol. 105, 2771–80.

    Article  PubMed  Google Scholar 

  • Williams, P. E. &Goldspink, G. (1971) Longitudinal growth of striated muscle fibres.J. Cell Sci. 9, 751–67.

    PubMed  Google Scholar 

  • Zak, R., Martin, A. F. &Blough, R. (1979) Assessment of protein turnover by use of radioisotopic tracers.Physiol. Rev. 59, 407–47.

    PubMed  Google Scholar 

  • Zak, R., Martin, A. F., Prior, G. &Rabinowitz, M. (1977) Comparison of turnover of several myofibrillar proteins and critical evaluation of double isotope method.J. Biol. Chem. 252, 3430–5.

    PubMed  Google Scholar 

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Franchi, L.L., Murdoch, A., Brown, W.E. et al. Subcellular localization of newly incorporated myosin in rabbit fast skeletal muscle undergoing stimulation-induced type transformation. J Muscle Res Cell Motil 11, 227–239 (1990). https://doi.org/10.1007/BF01843576

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