Summary
The ultrastructural organization of the highly interconnected filamentous network underneath the sarcolemma as well as the role played by the muscle protein dystrophin within this cytoskeleton remain yet unclear. More accurate information has been obtained by using a method which provides three-dimensional en face views of large membrane areas applied to mouse cultured myotubes and isolated adult skeletal muscle fibres. Two levels have been distinguished in the cytoskeleton underlying the sarcolemma: the submembranous level, partly integrated into the membrane, and the cortical level, invading the proximal cytoplasmic space. Few differences have been found between the membrane cytoskeletons of myotubes issued from 14-day-old cultures and those of adult fibres. The comparison was done with cells where dystrophin is missing (mdx mouse muscle): surprisingly, the lack of dystrophin does not induce obvious or dramatic ultrastructural disorganization, either in the cortical cytoskeletal network or in the submembranous one. Immunogold labelling of either the central-rod or the C-terminal domain of dystrophin is not located among the cortical network. This study provides additional data on the spatial ordering of subsarcolemmal cytoskeletal elements: dystrophin does not appear as a filamentous structure entirely located among subsarcolemmal cytoskeleton but seems partly embedded in membranous material.
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AGGELER, J. & WERB, Z. (1982) Initial events during phagocytosis by macrophages viewed from outside and inside the cell: membrane-particle interactions and clathrin. J. Cell Biol. 94, 613–23.
BATTEN, B. E., AALBERG, J. J. & ANDERSON, E. (1980) The cytoplasmic filamentous network in cultured ovarian granulosa cells. Cell 21, 885–95.
BERTHIER, C., AMSELLEM, J. & BLAINEAU, S. (1992) Organization of the cytoskeleton of dystrophin deficient mdx mouse myocytes-electron microscopic study. Biol. Cell 75, 7a.
BERTHIER, C., AMSELLEM, J. & BLAINEAU, S. (1994) En face views of subsarcolemmal cytoskeleton network of mouse muscle fibres: immunoelectron localization of dystrophin. 9th European Cytoskeleton Forum, 7th–12th September 1994, Dundee, UK P1, Poster abstract 1.
BYERS, T. J., KUNKEL, L. M. & WATKINS, S. C. (1991) The subcellular distribution of dystrophin in mouse skeletal, cardiac, and smooth muscle. J. Cell Biol. 115, 411–21.
CARPENTER, S., KARPATI, G., ZUBRZYCKA-GAARN, E., BULMAN, D. E., RAY, P. N. & WORTON, R. G. (1990) Dystrophin is localized to the plasma membrane of human skeletal muscle fibres by electron microscopic cytochemical study. Muscle & Nerve 13, 376–80.
CULLEN, M. J., WALSH, J., NICHOLSON, L. V. B. & HARRIS, J. B. (1990) Ultrastructural localization of dystrophin in human muscle by using gold immunolabelling. Proc. R. Soc. Lond. B 240, 197–210.
CULLEN, M. J., WALSH, J., NICHOLSON, L. V. B., HARRIS, J. B., ZUBRZYCKA-GAARN, E. E., RAY, P. N. & WORTON, R. G. (1991) Immunogold labelling of dystrophin in human muscle using an antibody to the last 17 amino acids of the C-terminus. Neuromusc Disord 1, 113–19.
DePriester, W. (1991) Techniques for the visualisation of cytoskeletal components in Dyctyostelium discoideum. Electron Microsc. Rev. 4, 343–76.
ERVASTI, J. M. & CAMPBELL, K. P. (1991) Membrane organization of the dystrophin-glycoprotein complex. Cell 66, 1121–31.
ERVASTI, J. M. & CAMPBELL, K. P. (1993) Dystrophin and the membrane skeleton. Curr. Opin. Cell Biol. 5, 82–7.
ERVASTI, J. M., OLHENDIECK, K., KAHL, S. D., GAVER, M. G. & CAMPBELL, K. P. (1990) Deficiency of a glycoprotein component of the dystrophin complex in dystrophic muscle. Nature 345, 315–19.
FABBRIZIO, E., HARRICANE, M. C., PONS, F., LEGER, J. & MORNET, D. (1992) Properties of chicken cardiac dystrophin. Biol. Cell 76, 167–74.
FABBRIZIO, E., LEGER, J., ANOAL, M., LEGER, J. J. & MORNET, D. (1993) Monoclonal antibodies targeted against the C-terminal domain of dystrophin or utrophin. FEBS Lett. 322, 10–14.
GINJAAR, H. B., VanPaassen, H. B. M., DenDunnen, J. T., NGUYEN THI, Man, MORRIS, G. E., MOORMAN, A. F. M. & VanOmmen, G. J. B. (1992) Construction of dystrophin fusion proteins to raise targeted antibodies to different epitopes. FEBS Lett. 308, 293–7.
HOFFMAN, E. P., BROWN, R. H. J. & KUNKEL, L. M. (1987) Dystrophin: the protein product of the Duchenne muscular dystrophy locus. Cell 51, 919–28.
HOFFMAN, E. P., MORGAN, J. E., WATKINS, S. C. & PARTRIDGE, T. A. (1990) Somatic reversion/suppression of the mouse mdx phenotype in vivo. J. Neurol. Sci. 99, 9–25.
HUARD, J., LARBRECQUE, C., DANSEREAU, G., ROBITAILLE, L. & TREMBLAY, J. P. (1991) Dystrophin expression in myotubes formed by the fusion of normal and dystrophic myoblasts. Muscle & Nerve 14, 178–82.
IBRAGHIMOV-BESKROVNAYA, O., ERVASTI, J. M., LEVEILLE, C. J., SLAUGHTER, C. A., SERNETT, S. W. & CAMPBELL, K. P. (1992) Primary structure of dystrophin-associated glycoproteins linking dystrophin to the extracellular matrix. Nature 355, 696–702.
ISOBE, Y. & SHIMADA, Y. (1986) Organization of filaments underneath the plasma membrane of developing chicken skeletal muscle cells in vitro revealed by the freeze-dry and rotary replica method. Cell Tissue Res. 244, 47–56.
KOENIG, M., MONACO, A. P. & KUNKEL, L. M. (1988) The complete sequence of dystrophin predicts a rodshaped cytoskeletal protein. Cell 53, 219–28.
KÜHL, U., Öcalan, M., TIMPL, R. & Von DerMark, K. (1986) Role of laminin and fibronectin in selecting myogenic versus fibrogenic cells from skeletal muscle cells in vitro. Dev. Biol. 117, 628–35.
LAZARIDES, E. (1980) Intermediate filaments as mechanical integrators of cellular space. Nature 283, 249–56.
LIU, S. C., DERICK, L. H. & PALEK, J. (1987) Visualization of the hexagonal lattice in the erythrocyte membrane skeleton. J. Cell Biol. 104, 527–36.
LUNA, E. J. & HITT, A. L. (1992) Cytoskeleton-plasma membrane interactions. Science 258, 955–63.
MASSA, R., CASTELLANI, L., SILVESTRI, G., SANCESARIO, G. & BERNARDI, G. (1994) Dystrophin is not essential for the integrity of the cytoskeleton. Acta Neuropathol. 87, 377–84.
MASUDA, T., FUJIMAKI, N., OZAWA, E. & ISHIKAWA, H. (1992) Confocal laser microscopy of dystrophin localization in guinea pig skeletal muscle fibers. J. Cell Biol. 119, 543–8.
MATSUMURA, K., ERVASTI, J. M., OHLENDIECK, K., KAHL, S. D. & CAMPBELL, K. P. (1992) Association of dystrophin-related protein with dystrophin-associated proteins in mdx mouse muscle. Nature 360, 588–91.
MAZIA, D., SCHATTEN, G. & SALE, W. (1975) Adhesion of cells to surfaces coated with polylysine-applications to electron microscopy. J. Cell Biol. 66, 198–200.
NERMUT, M. V. (1989) Strategy and tactics in electron microscopy of cell surfaces. Electron. Microsc. Rev. 2, 171–96.
NERMUT, M. V. & NICOL, A. (1989) Colloidal gold immunoreplica method. In Colloidal gold: principles, methods and applications vol. 1 (edited by Hayat, M. A.) pp. 349–73, London: Academic Press.
PARDO, J. V., SILICIANO, J. D. & CRAIG, S. W. (1983) A vinculin-containing cortical lattice in skeletal muscle: transverse lattice elements (‘costameres’) mark sites of attachment between myofibrils and the sarcolemma. Proc. Natl. Acad. Sci. USA 80, 1008–12.
PIEROBON-BORNIOLI, S. (1981) Transverse sarcomere filamentous network of myofibril-sarcolemma attachment regions in cardiac muscle fibers. J. Cell Biol. 97, 1081–8.
PONS, J., AUGIER, N., HEILIG, R., LEGER, J., MORNET, D. & LEGER, J. J. (1990) Isolated dystrophin molecules as seen by electron microscopy. Proc. Natl. Acad. Sci. USA 87, 7851–5.
PORTER, G. A., DMYTRENKO, G. M., WINKELMANN, J. C. & BLOCH, R. J. (1992) Dystrophin colocalizes with betaspectrin in distinct subsarcolemmal domains in mammalian skeletal muscle. J. Cell Biol. 117, 997–1005.
SABBADINI, R. A. & DAHMS, A. S. (1989) Biochemical properties of isolated transverse tubular membranes. J. Bioenerg. Biomembr. 21, 163–213.
SATO, O., NONOMURA, Y., KIMURA, S. & MARUYAMA, K. (1992) Molecular shape of dystrophin. J. Biochem. 112, 631–6.
SHEAR, C. R. & BLOCH, R. J. (1985) Vinculin in subsarcolemmal densities in chicken skeletal muscle: localization and relationship to intracellular and extracellular structures. J. Cell Biol. 101, 240–56.
SMALL, J. V., FURST, D. O. & THORNELL, L. E. (1992) The cytoskeletal lattice of muscle cells. Eur. J. Biochem. 208, 559–72.
SQUARZONI, S., SABATELLI, P., MALTARELLO, M. C., CATALDI, A., DiPrimio, R. & MARLDI, N. M. (1992) Localization of dystrophin COOH-terminal domain by the fracture-label technique. J. Cell Biol. 118, 1401–9.
STRAUB, V., BITTNER, R. E., LEGER, J. J. & VOIT, T. (1992) Direct visualization of the dystrophin network on skeletal muscle fiber membrane. J. Cell Biol. 119, 1183–91.
THORNELL, L. E. & PRICE, M. G. (1991) The cytoskeleton in muscle cells in relation to function. Bioch. Soc. Transac. 19, 1116–20.
WAKAYAMA, Y. & SHIBUYA, S. (1990) Observations on the muscle plasma membrane-associated cytoskeletons of mdx mice by quick-freeze, deep-etch, rotary-shadow replica method. Acta Neuropathol. 80, 618–23.
WAKAYAMA, Y. & SHIBUYA, S. (1991a) Gold-labelled dystrophin molecule in muscle plasmalemma of mdx control mice as seen by electron microscopy of deep etching replica. Acta Neuropathol. 82, 178–84.
WAKAYAMA, Y. & SHIBUYA, S. (1991b) Antibody-decorated dystrophin molecule of murine skeletal myofiber as seen by freeze-etching electron microscopy. J. Electron. Microsc. 40, 143–5.
WAKAYAMA, Y., SHIBUYA, S., JIMI, T., TAKEDA, A. & ONIKI, H. (1993) Size and localization of dystrophin molecule-immunoelectron microscopic and freeze etching studies of muscle plasma membranes of murine skeletal myofibers. Acta Neuropathol. 86, 567–77.
WAKAYAMA, Y., SHIBUYA, S., TAKEDA, A., JIMI, T., NAKAMURA, Y. & ONIKI, H. (1995) Ultrastructural localization of the C-terminus of the 43-kd dystrophin-associated glycoprotein and its relation to dystrophin in normal murine skeletal myofiber. Am. J. Pathol. 146, 189–96.
WATKINS, S. C., HOFFMAN, E. P., SLAYTER, H. S. & KUNKEL, L. M. (1988) Immunoelectron microscopic localization of dystrophin in myofibres. Nature 333, 863–6.
YURCHENCO, P. D. & RUBEN, G. C. (1987) Basement membrane structure in situ: Evidence for lateral associations in the type IV collagen network. J. Cell Biol. 105, 2559–68.
ZUBRZYCKA-GAARN, E. E., HUTTER, O. F., KARPATI, G., KLAMUT, H. J., BULMAN, D. E., HODGES, R. S., WORTON, R. G. & RAY, P. N. (1991) Dystrophin is tightly associated with the sarcolemma of mammalian skeletal muscle fibers. Exp. Cell Res. 192, 278–88.
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Berthier, C., Amsellem, J. & Blaineau, S. Visualization of the subsarcolemmal cytoskeleton network of mouse skeletal muscle cells by en face views and application to immunoelectron localization of dystrophin. J Muscle Res Cell Motil 16, 553–566 (1995). https://doi.org/10.1007/BF00126439
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DOI: https://doi.org/10.1007/BF00126439