Skip to main content
Log in

Myosin filaments isolated from skinned amphibian smooth muscle cells are side-polar

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

Summary

The structure of myosin filaments isolated from skinned toad stomach smooth muscle cells has been examined by electron microscopy as a step toward identifying thein vivo structure. When negatively stained following exposure to relaxing conditions, the filaments exhibited a continuous 14-nm axial repeat of crossbridge projections with no central bare zone. The filaments thus differed from the bipolar filaments found in striated muscle and displayed instead features resembling side-polar and mixed-polarity filament models. By rotation of isolated filaments around their longitudinal axes it was found that cross bridges occurred only along two sides of the filament, an arrangement consistent with the side-polar but not the mixed-polarity model. The polarity is thus similar to that proposed for ribbons (Small & Squire,J. molec. Biol. 67, (1972) 17–149) and for synthetic smooth muscle myosin filaments (Craig and Megerman,J. Cell Biol. 75, (1977) 990–996); their appearance in cross-section, however, shows that these structures are filaments (i.e. with two axes of similar dimensions) and not broad ribbons. As the filaments were derived directly from skinned cells which contracted and relaxed in response to physiological levels of MgATP and Ca2+ at rates comparable to those of native, isolated cells, this unusual arrangement of cross bridges appears to be an effective, functional form of myosin in the contractile apparatus. Side-polar filaments therefore merit consideration as plausible candidates for the native organization of myosin in vertebrate smooth muscle cells.

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

  • Ashton, F. T., Somlyo, A. V. &Somlyo, A. P. (1975) The contractile apparatus of vascular smooth muscle: intermediate high voltage stereo electron microscopy.J. molec. Biol. 98, 17–29.

    Google Scholar 

  • Bagby, R. (1986) Towards a comprehensive three-dimensional model of the contractile system of vertebrate smooth muscle cells.Int. Rev. Cytol. 105, 67–128.

    Google Scholar 

  • Bagby, R. M., Young, A. M., Dotson, R. S., Fisher, B. A. &McKinnon, K. (1971) Contraction of single smooth muscle cells from Bufo marinus stomach.Nature 234, 351–2.

    Google Scholar 

  • Bagby, R. M. &Fisher, B. A. (1979) Effects of DMSO, pH, stretch and calcium on the thick filaments in an amphibian smooth muscle.Eur. J. Cell Biol. 19, 196–200.

    Google Scholar 

  • Bond, M. &Somlyo, A. v. (1982) Dense bodies and actin polarity in vertebrate smooth muscle.J. Cell Biol. 95, 403–13.

    Google Scholar 

  • Buhle, E. L., Somlyo, A. V. &Somlyo, A. P. (1986) A comparison of isolated andin situ thick filaments from smooth muscle.Proc. Ann. EMSA 44, 156–7.

    Google Scholar 

  • Cande, W. Z., Tooth, P. J. &Kendrick-Jones, J. (1983) Regulation of contraction and thick filament assembly-disassembly in glycerinated vertebrate smooth muscle cells.J. Cell Biol. 97, 1062–71.

    Google Scholar 

  • Cooke, P. (1982) A reversible change in the functional organization of thin filaments in smooth muscle fibres.Eur. J. Cell Biol. 27, 55–61.

    Google Scholar 

  • Cooke, P. (1983) Organization of contractile fibers in smooth muscle. InCell and Muscle Motility (edited byDowben, R. M. AndShay, J. W.), Vol. 3, pp. 57–77. New York: Plenum.

    Google Scholar 

  • Cooke, P. H. &Fay, F. S. (1975) Correlation between fibre length, ultrastructure and the length-tension relationship of mammalian smooth muscle.J. Cell Biol. 52, 105–16.

    Google Scholar 

  • Craig, R. &Megerman, J. (1977) Assembly of smooth muscle myosin into side-polar filaments.J. Cell Biol. 75, 990–6.

    Google Scholar 

  • Craig, R. &Megerman, J. (1979) Electron microscope studies on muscle thick filaments. InMotility in Cell Function (edited byPepe, F. A., Sanger, J. W. AndNachmias, V. T.) pp. 91–102. New York: Academic Press.

    Google Scholar 

  • Fay, F. S. &Singer, J. J. (1977) Characteristics of response of isolated smooth muscle cells to cholinergic drugs.Am. J. Physiol. 232, C144-C154.

    Google Scholar 

  • Fay, F. S., Fujiwara, K., Rees, D. D. &Fogarty, K. E. (1983) Distribution of α-actinin in single isolated smooth muscle cells.J. Cell Biol. 96, 783–95.

    Google Scholar 

  • Fay, F. S., Hoffman, R., Leclair, S. &Merriam, P. (1982) Preparation of individual smooth muscle cells from the stomach ofBufo marinus.Meth. Enzym. 85, 284–92.

    Google Scholar 

  • Gillis, J. M., Cao, M. Y. &Godfraind-De Becker, A. (1988) Density of myosin filaments in the rat anococcygeus muscle, at rest and in contraction. II.J. Musc. Res. Cell Motility 9, 18–28.

    Google Scholar 

  • Godfraind-De Becker, A. &Gillis, J. M. (1988) Analysis of the birefringence of the smooth muscle anococcygeus of the rat, at rest and in contraction. I.J. Musc. Res. Cell Motility 9, 9–17.

    Google Scholar 

  • Hinssen, H., D'Haese, J., Small, J. V. &Sobieszek, A. (1978) Mode of filament assembly of myosins from muscle and non-muscle cells.J. Ultrastruct. Res. 64, 282–302.

    Google Scholar 

  • Huxley, H. E. (1963) Electron microscope studies on the structure of natural and synthetic protein filaments from striated muscle.J. molec. Biol. 7, 281–308.

    Google Scholar 

  • Huxley, H. E. &Brown, W. (1967) The low-angle X-ray diagram of vertebrate striated muscle and its behaviour during contraction and rigor.J. molec. Biol. 30, 383–434.

    Google Scholar 

  • Kaminer, B. (1969) Synthetic myosin filaments from vertebrate smooth muscle.J. molec. Biol. 39, 257–64.

    Google Scholar 

  • Kargacin, G. J. &Fay, F. S. (1987) Physiological and structural properties of saponin-skinned single smooth muscle cells.J. gen. Physiol. 90, 49–73.

    Google Scholar 

  • Kelly, R. E. &Rice, R. V. (1968) Localization of myosin filaments in smooth muscle.J. Cell Biol. 37, 105–16.

    Google Scholar 

  • Kendrick-Jones, J., Smith, R. C., Craig, R. &Citi, S. (1987) Polymerization of vertebrate non-muscle and smooth muscle myosins.J. molec. Biol. 198, 241–52.

    Google Scholar 

  • Lowy, J. &Small, J. v. (1970) The organization of myosin and actin in vertebrate smooth muscle.Nature 227, 1–6.

    Google Scholar 

  • Reedy, M. K. (1968) Ultrastructure of insect flight muscle. I. Screw sense and structural grouping in the rigor crossbridge lattice.J. molec. Biol. 31, 155–76.

    Google Scholar 

  • Reedy, M. K., Holmes, K. C. &Tregear, R. T. (1965) Induced changes in orientation of the cross-bridges of glycerinated insect flight muscle.Nature 207, 1276–80.

    Google Scholar 

  • Salmon, E. D. &Derosier, D. (1981) A surveying optical diffractometer.J. Microsc. 123, 239–47.

    Google Scholar 

  • Scheid, C. R., Honeyman, T. W. &Fay, F. S. (1979) The mechanism of Β-adrenergic relaxation of smooth muscle.Nature 277, 32–6.

    Google Scholar 

  • Scholey, J. M., Taylor, K. A. &Kendrick-Jones, J. (1981) The role of myosin light chains in regulating actin-myosin interactions.Biochimie (Paris) 63, 255–71.

    Google Scholar 

  • Shoenberg, C. F. (1969) An electron miscroscope study of the influence of divalent ions on myosin filament formation in chicken gizzard extracts and homogenates.Tissue Cell 1, 83–96.

    Google Scholar 

  • Shoenberg, C. F. &Needham, D. M. (1976) A Study of the mechanism of contraction in vertebrate smooth muscle.Biol. Rev. 51, 53–104.

    Google Scholar 

  • Small, J. V. (1977) Studies on isolated smooth muscle cells: the contractile apparatus.J. Cell Sci. 24, 327–49.

    Google Scholar 

  • Small, J. V. &Squire, J. M. (1972) Structural basis of contraction in vertebrate smooth muscle.J. molec. Biol. 67, 117–49.

    Google Scholar 

  • Somlyo, A. P., Somlyo, A. V., Devine, C. E. &Rice, R. V. (1971) Aggregation of thick filaments into ribbons in mammalian smooth muscle.Nature New Biol. 231, 243–6.

    Google Scholar 

  • Somlyo, A. P., Devine, C. E., Somlyo, A. V. &Rice, R. V. (1973) Filament organization in vertebrate smooth muscle.Phil. Trans. R. Soc. Lond. B265, 223–9.

    Google Scholar 

  • Somlyo, A. V., Butler, T. M., Bond, M. &Somlyo, A. P. (1981) Myosin filaments have nonphosphorylated light chains in relaxed smooth muscle.Nature 294, 567–9.

    Google Scholar 

  • Somlyo, A. v. (1980) Ultrastructure of vascular smooth muscle. InHandbook of Physiology (edited byBohr, D., Somlyo, A. andSparks, H.), Section 2, Vol. 2, pp. 33–67. Bethesda: American Physiological Society.

    Google Scholar 

  • Squire, J. M. (1975) Muscle filament structure and muscle contraction.Ann. Rev. Biophys. Bioeng. 4, 137–63.

    Google Scholar 

  • Stafford, W. F. (1985) Effect of various anions on the stability of the coiled-coil of skeletal muscle myosin.Biochemistry 24, 3314–21.

    Google Scholar 

  • Suzuki, H., Onishi, H., Takahashi, K. &Watanabe, S. (1978) Structure and function of chicken gizzard myosin.J. Biochem (Tokyo) 84, 1529–42.

    Google Scholar 

  • Trybus, K. M. &Lowey, S. (1987a) Subunit exchange between smooth muscle myosin filaments.J. Cell Biol. 105, 3021–30.

    Google Scholar 

  • Trybus, K. M. &Lowey., S. (1987b) Assembly of smooth muscle myosin minifilaments: effects of phosphorylation and nucleotide binding.J. Cell. Biol. 105, 3007–19.

    Google Scholar 

  • Warshaw, D. M. &Fay, F. S. (1983) Cross-bridge elasticity in single smooth muscle cells.J. gen. Physiol. 82, 157–99.

    Google Scholar 

  • Wray, J. S., Vibert, P. J. &Cohen, C. (1975) Diversity of cross-bridge configurations in invertebrate muscles.Nature 257, 561–4.

    Google Scholar 

  • Wray, J., Vibert, P. &Cohen, C. (1978) Actin filaments in muscle: pattern of myosin and tropomyosin/troponin attachments.J. molec. Biol. 124, 501–22.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cooke, P.H., Fay, F.S. & Craig, R. Myosin filaments isolated from skinned amphibian smooth muscle cells are side-polar. J Muscle Res Cell Motil 10, 206–220 (1989). https://doi.org/10.1007/BF01739811

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01739811

Keywords

Navigation