Skip to main content
Log in

Three myosin heavy chain isoforms in type 2 skeletal muscle fibres

  • Published:
Journal of Muscle Research & Cell Motility Aims and scope Submit manuscript

Summary

Mammalian skeletal muscles consist of three main fibre types, type 1, 2A and 2B fibres, with different myosin heavy chain (MHC) composition. We have now identified another fibre type, called type 2X fibre, characterized by a specific MHC isoform. Type 2X fibres, which are widely distributed in rat skeletal muscles, can be distinguished from 2A and 2B fibres by histochemical ATPase activity and by their unique staining pattern with seven anti-MHC monoclonal antibodies. The existence of the 2X-MHC isoform was confirmed by immunoblotting analysis using muscles containing 2X fibres as a major component, such as the normal and hyperthyroid diaphragm, and the soleus muscle after high frequency chronic stimulation. 2X-MHC contains one determinant common to 2B-MHC and another common to all type 2-MHCs, but lacks epitopes specific for 2A- and 2B-MHCs, as well as an epitope present on all other MHCs. By SDS-polyacrylamide gel electrophoresis 2X-MHC shows a lower mobility compared to 2B-MHC and appears to comigrate with 2A-MHC. Muscles containing predominantly 2X-MHC display a velocity of shortening intermediate between that of slow muscles and that of fast muscles composed predominantly of 2B fibres.

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.

Similar content being viewed by others

References

  • BÄr, A. &Pette, D. (1988) Three fast myosin heavy chains in adult rat skeletal muscle.FEBS Lett. 235, 153–5.

    Google Scholar 

  • Barany, M. (1967) ATPase activity of myosin correlated with speed of muscle shortening.J. gen. Physiol. 50, 197–216.

    Google Scholar 

  • Billeter, R., Heizmann, C. W., Howard, H. &Jenny, E. (1981) Analysis of myosin light and heavy chain types in single human skeletal muscle fibres.Eur. J. Biochem. 116, 389–95.

    Google Scholar 

  • Brooke, M. H. &Kaiser, K. K. (1970) Muscle fiber types: how many and what kind?Arch. Neurol. 23, 369–79.

    Google Scholar 

  • Burke, R. E., Levine, D. N., Tsairis, P. &Zajac, F. E. (1973) Physiological types and histochemical profiles in motor units of the cat gastrocnemius.J. Physiol. (Lond.) 234, 723–48.

    Google Scholar 

  • Dalla Libera, L., Sartore, S., Pierobon-Bormioli, S. &Schiaffino, S. (1980) Fast-white and fast-red isomyosins in guinea pig muscles.Biochem. Biophys. Res. Commun. 96, 1662–70.

    Google Scholar 

  • Danielibetto, D., Zerbato, E. &Betto, R. (1986) Type 1, 2A and 2B myosin heavy chain electrophoretic analysis of rat muscle fibers.Biochem. Biophys. Res. Commun. 138, 981–7.

    Google Scholar 

  • Dhoot, G. K., Hales, M. C., Grail, B. M. &Perry, S. V. (1985) The isoforms of C protein and their distribution in mammalian skeletal muscle.J. Musc. Res. Cell Motility 6, 487–505.

    Google Scholar 

  • Eddinger, T. J. &Moss, R. L. (1987) Mechanical properties of skinned single fibers of identified types from rat diaphragm.Am. J. Physiol. 253, C210–8.

    Google Scholar 

  • Edstrom, L. &Kugelberg, E. (1968) Histochemical composition, distribution of fibres and fatiguability of single motor units.J. Neurol. Neurosurg. Psychiat. 31, 424–33.

    Google Scholar 

  • Gorza, L., Sartore, S., Thornell, L. E. &Schiaffino, S. (1986) Myosin types and fiber types in cardiac muscle. III. Nodal conduction tissue.J. cell. Biol. 102, 1758–66.

    Google Scholar 

  • Gorza, L., Gundersen, K., LØmo, T., &Schiaffino, S. (1988) Slow-to-fast transformation of denervated soleus muscles by chronic high-frequency stimulation in the rat.J. Physiol. (Lond.) 402, 627–49.

    Google Scholar 

  • Guth, L. &Samaha, F. J. (1969) Qualitative differences between actomyosin ATPase of slow and fast mammalian muscle.Exp. Neurol. 25, 138–52.

    Google Scholar 

  • Hintz, C., Coyle, E. F., Kaiser, K. K., Chi, M. M. Y. &Lowry, O. H. (1984) Comparison of muscle fiber typing by quantitative enzyme assays and by myosin ATPase staining.J. Histochem. Cytochem. 32, 655–60.

    Google Scholar 

  • Hoh, J. F. Y., McGrath, P. A. &Hale, P. T. (1978) Electrophoretic analysis of multiple forms of rat cardiac myosin: Effect of hypophysectomy and thyroxine replacementJ. molec. cell. Cardiol. 10, 1053–76.

    Google Scholar 

  • Hudson, L. &Hay, F. C. (1980)Practical Immunology, 2nd edn pp. 220–222. Oxford: Blackwell Scientific.

    Google Scholar 

  • Izumo, S., Nadal-Ginard, B. &Mahdavi, V. (1986) All members of the myosin heavy chain multi-gene family respond to thyroid hormone in a highly tissue specific manner.Science 231, 597–600.

    Google Scholar 

  • Julian, F. J., Moss, R. L., &Waller, G. S. (1981) Mechanical properties and myosin light chain composition of skinned muscle fibres from adult and newborn rats.J. Physiol. (Lond.) 311, 201–18.

    Google Scholar 

  • Kugelberg, E. &Lindegren, B. (1979) Transmission and contraction fatigue of rat motor units in relation to succinate dehydrogenase activity of motor unit fibres.J. Physiol. (Lond.) 288, 285–300.

    Google Scholar 

  • Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature 227, 680–5.

    Google Scholar 

  • Lannergren, J. (1987) Contractile properties and myosin isoenzymes of various kinds ofXenopus twitch muscle fibres.J. Musc. Res. Cell Motility 8, 260–73.

    Google Scholar 

  • Leung, B., Kula, R. W. &Shafiq, S. A. (1987) Fiber types in normal and neonatally denervated fast muscles of the rat: immunocytochemical study with an antimyosin monoclonal antibody.Exp. Neurol. 97, 429–40.

    Google Scholar 

  • Luff, A. R. (1981) Dynamic properties of the inferior recrus, extensor digitorum longus, diaphragm and soleus muscles of the mouse.J. Physiol. (Lond.) 313, 161–71.

    Google Scholar 

  • Mabuchi, K., Mabuchi, Y., Sreter, F. A. &Gergely, J. (1988) Subdivision of leg type 2B fibres in rabbit and rat skeletal muscles.Biophys. J. 53, 179a.

    Google Scholar 

  • Margossian, S. S. &Lowey, S. (1982) Preparation of myosin and its subfragments from rabbit skeletal muscle.Meth. Enzymol. 85, 55–71.

    Google Scholar 

  • Moore, G. E. &Schachat, F. H. (1985) Molecular heterogeneity of histochemical fibre types: a comparison of fast fibres.J. Musc. Res. Cell Motility 6, 513–24.

    Google Scholar 

  • Morrissey, J. H. (1981) Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity.Analyt. Biochem. 117, 307–10.

    Google Scholar 

  • Nachlas, M. M., Tsou, K. G., De Sousa, E., Cheng, C. S. &Seligman, A. M. (1957) Cytochemical demonstration of succinic dehydrogenase by the use of a newp-nitrophenyl substituted ditetrazolium.J. Histochem. Cytochem. 5, 420–36.

    Google Scholar 

  • Nemeth, P. &Pette, D. (1981) Succinate dehydrogenase activity in fibres classified by myosin ATPase in three hind limb muscles of rat.J. Physiol. (Lond.) 320, 73–80.

    Google Scholar 

  • Pagani, E. D. &Julian, F. J. (1984) Rabbit papillary muscle myosin isozymes and the velocity of muscle shortening.Circ. Res. 54, 586–94.

    Google Scholar 

  • Pierobon-Bormioli, S., Sartore, S., Dalla Libera, L., Vitadello, M. &Schiaffino, S. (1981) Fast isomyosins and fiber types in mammalian skeletal muscle.J. Histochem. Cytochem. 29, 1179–88.

    Google Scholar 

  • Reiser, P. J., Moss, R. L., Giulian, G. G. &Greaser, M. L. (1985) Shortening velocity in single fibers from adult rabbit soleus muscles is correlated with myosin heavy chain composition.J. biol. Chem. 260, 9077–80.

    Google Scholar 

  • Russo, C., Callegaro, L., Lanza, L. &Ferrone, S. (1983) Purification of IgG monoclonal antibodies by caprylic acid technique.J. Immun. Meth. 65, 269–71.

    Google Scholar 

  • Sartore, S., Gorza, L. &Schiaffino, S. (1982) Fetal myosin heavy chain in regenerating muscle.Nature 298, 294–6.

    Google Scholar 

  • Sartore, S., Mascarello, F., Rowlerson, A., Gorza, L., Ausoni, S., Vianello, M. &Schiaffino, S. (1987) Fibre types in extraocular muscles: A new myosin isoform in the fast fibres.J. Musc. Res. Cell Motility 8, 161–71.

    Google Scholar 

  • Schachat, F. H., Bronson, D. D. &McDonald, O. B. (1985a) Heterogeneity of contractile proteins. A continuum of troponin-tropomyosin expression in mammalian skeletal muscle.J. biol. Chem. 260, 1108–13.

    Google Scholar 

  • Schachat, F. H., Canine, A. C., Briggs, M. M. &Reedy, M. C. (1985b) The presence of two skeletal muscle α-actinins correlates with troponin-tropomyosin expression and Z-line width.J. cell Biol. 101, 1001–8.

    Google Scholar 

  • Schiaffino, S., Ausoni, S., Gorza, L., Saggin, L., Gundersen, K., &LØmo, T. (1988a) Myosin heavy chain isoforms and velocity of shortening of type 2 skeletal muscle fibres.Acta Physiol. Scand. 134, 565–6.

    Google Scholar 

  • Schiaffino, S., Gorza, L., Pitton, G., Saggin, L., Ausoni, S., Sartore, S. &LØmo, T. (1988b) Embryonic and neonatal myosin heavy chain in denervated and paralysed rat skeletal muscle.Devl. Biol. 127, 1–11.

    Google Scholar 

  • Schiaffino, S., Gorza, L., Sartore, S., Saggin, L. &Carli, M. (1986a) Embryonic myosin heavy chain as a differentiation marker of human developing muscle and rhabdomyosarcoma. A monoclonal antibody study.Exp. Cell Res. 163, 211–20.

    Google Scholar 

  • Schiaffino, S., Hanzlikova, V. &Pierobon, S. (1970) Relation between structure and function in rat skeletal muscle fibres.J. cell Biol. 47, 107–19.

    Google Scholar 

  • Schiaffino, S., Saggin, L., Viel, A., Ausoni, S., Sartore, S. &Gorza, L. (1986b) Muscle fibre types identified by monoclonal antibodies to myosin heavy chains. InBiochemical Aspects of Physical Exercise (edited byBenzi, G., Packer, L. &Siliprandi, N.), pp. 27–34, Amsterdam: Elsevier.

    Google Scholar 

  • Schiaffino, S., Saggin, L., Viel, A., &Gorza, L. (1985) Differentiation of fibre types in rat skeletal muscle visualized with monoclonal antimyosin antibodies.J. Musc. Res. Cell Motility 6, 60–1.

    Google Scholar 

  • Schwartz, K., Lecarpentier, Y., Martin, J. L., Lompré, A. M., Mercadier, J. J. &Swynghedauw, B. (1981) Myosin isoenzymic distribution correlates with speed of myocardial contraction.J. molec. cell. Cardiol. 13, 1071–5.

    Google Scholar 

  • Starr, R., Almond, R. &Offer, G. (1985) Location of C-protein, H-protein and X-protein in rabbit skeletal muscle fibre types.J. Musc. Res. Cell Motility 6, 227–56.

    Google Scholar 

  • Sweeney, H. L., Kushmerick, M. J., Mabuchi, K., Gergely, J. &Sreter, F. A. (1986) Velocity of shortening and myosin isozymes in two types of rabbit fast-twitch muscle fibres.Am. J. Physiol. 251, C431–4.

    Google Scholar 

  • Sweeney, H. L., Kushmerick, M. J., Mabuchi, K., Sreter, F. A. &Gergely, J. (1988) Myosin alkali light chain and heavy chain variations correlate with altered shortening velocity of isolated skeletal muscle fibres.J. biol. Chem. 263, 9034–9.

    Google Scholar 

  • Towbin, H., Staehelin, T. &Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proc. natn. Acad. Sci. U.S.A. 76, 4350–4.

    Google Scholar 

  • Trojanowski, J. Q., Obrocka, M. A. &Lee, V. M. Y. (1983) A comparison of eight different chromogen protocols for the demonstration of immunoreactive neurofilaments in rat cerebellum using the peroxidase-antiperoxidase method and monoclonal antibodies.J. Histochem. Cytochem. 31, 1217–23.

    Google Scholar 

  • Wagner, P. D. (1981) Formation and characterization of myosin hybrids containing essential light chains and heavy chains from different muscle myosins.J. biol. Chem. 256, 2493–8.

    Google Scholar 

  • Wieczorek, D. F., Periasamy, M., Butler-Browne, G. S., Whalen, R. G. &Nadal-Ginard, B. (1985) Co-expression of multiple myosin heavy chain genes, in addition to a tissue-specific one,in extraocular musculature.J. cell. Biol. 101, 618–29.

    Google Scholar 

  • Zweig, S. E. (1981) The muscle specificity and structure of two closely related fast-twitch white muscle myosin heavy chain isozymes.J. biol. Chem. 256, 11847–53.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schiaffino, S., Gorza, L., Sartore, S. et al. Three myosin heavy chain isoforms in type 2 skeletal muscle fibres. J Muscle Res Cell Motil 10, 197–205 (1989). https://doi.org/10.1007/BF01739810

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01739810

Keywords

Navigation