Summary
We have used monoclonal antibodies to desmin and titin, and a combination of immunofluoescence and immunogold labelling to study the disposition of these two proteins in normal human muscle fibres and in fibres at various stages of degeneration in dystrophic muscle. The normal pattern of desmin labelling, in particular the subsarcolemmal labelling, became disrupted at an early stage of fibre breakdown. There was a change from a transverse to a longitudinal orientation of the labelled intermediate filaments as the myofibrils sheared relative to one another. Thus, while it is probable that the desmin filaments are able to play a role in the mechanical integration of the myofibrils in healthy muscle, our results suggest that they cannot withstand the excessive forces generated by the hypercontraction and stretching of dystrophic muscle. However, small accumulations of desmin persisted between the damaged myofibrils until necrosis reached an advanced stage. In general, the degradation of titin appeared to occur before the degradation of desmin, and at the ultrastructural level, labelling with antibodies to epitopes from parts of the titin molecule close to the A-I-band junction was lost before labelling with an antibody to an epitope in the A-band. This suggests that different regions of the titin molecule break down at different stages in the breakdown of the fibre. We propose that lysis of titin in the I-band may underlie ‘slippage’, an abnormality often seen in dystrophic muscle, in which the A-band slips to one pole of the sarcomere such that it abuts onto the Z-line. Breakdown of the A-band section of titin may facilitate the disassembly of the A-filaments.
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References
Cullen MJ, Fulthorpe JJ (1975) Stages in fibre breakdown in Duchenne muscular Dystrophy. An electron-microscopy study. J Neurol Sci 24:179–200
Cullen MJ, Mastaglia FL (1980) Morphological changes in dystrophic muscle. Br Med Bull 36:145–152
Cullen MJ, Walsh J, Nicholson LVB, Harris JB (1990) Ultrastructural localization of dystrophin in human muscle using gold immunolabelling. Proc R Soc Lond [B] 240:197–210
Engel AG (1986) Duchenne dystrophy. In: Engel AG, Banker BQ (eds) Myology. McGraw-hill, New York, pp 1185–1240
Fürst DO, Osborn M, Nave R, Weber K (1988) The organization of titin filaments in the half sarcomere revealed by monoclonal antibodies in immunoelectron microscopy. J Cell Biol 106:1563–1572
Fürst DO, Osborn M, Weber K (1989) Myogenesis in the mouse embryo. Differential onset of expression of myogenic proteins and the involvement of titin in myofibril assembly. J Cell Biol 109:517–527
Hill CS, Duran S, Lin Z, Weber K, Holtzer H (1986) Titin and myosin, but not desmin, are linked during myofibrillogenesis in postmitotic mononucleated myoblasts. J Cell Biol 103:2185–2196
Horowitz R, Podolsky RJ (1987) The positional stability of thick filaments in activated skeletal muscle depends on sarcomere length. Evidence for the role of titin filaments. J Cell Biol 105:2217–2223
Horowitz R, Kempner ES, Bisher ME, Podolsky RJ (1986) A physiological role for titin and nebulin in skeletal muscle. Nature 323:160–164
Itoh Y, Suzuki T, Kimura S, Ohashi K, Higuchi H, Sawada H, Shimizu T, Shibata M, Maruyama K (1988) Extensible and less-extensible domains of connectin filaments in stretched vertebrate skeletal muscle sarcomeres as detected by immunofluorescence and immunofluorescence and immunoelectron microscopy using monoclonal antibodies. J Biochem 104:504–508
Komiyama M, Maruyama K, Shimada Y (1990) Assembly of connectin (titin) in relation to myosin and α-actinin in cultured cardiac myocytes. J Musc Res Cell Motil 11:419–428
Lazarides E (1980) Intermediate filaments as mechanical integrators of cellular space. Nature 282:249–256
Lidov HGW, Byers TJ, Watkins SC, Kunkel LM (199) Localization of dystrophin to postsynaptic regions of central nervous system cortical neurons. Nature 348:725–728
Maruyama K, Sawada H, Kimura S, Ohashik, Higuchi H, Umazume Y (1984) Connectin filaments in stretched skinned fibres of frog skeletal muscle. J Cell Biol 99:1391–1397
Matsumara K, Shimizu T, Nonaka I, Mannen T (1989) Immunochemical study of connectin (titin) in neuromuscular diseases using a monoclonal antibody. Connectin is degraded extensively in Duchenne muscular dystrophy. J Neurol Sci 93:147–156
Mokri B, Engel AG (1975) Duchenne muscular dystrophy. Electron microscopic findings pointing to a basic or early abnormality in the plasma membrane of the muscle fibre. Neurology 25:1111–1119
Ohlendieck K, Ervasti JM, Snook JB, Campbell KP (1991) Dystrophin-glycoprotein complex is highly enriched in isolated skeletal muscle sarcolemma. J Cell Biol 112:135–148
Pierobon-Bormioli S, Betto R, Salviati G (1989) The organization of titin (connectin) and nebulin in the sarcomeres: an immunocytolocalization study. J Musc Res Cell Motil 10:446–456
Schmalbruch H (1975) Segmental fibre breakdown and defects of the plasmalemma in diseased human muscle. Acta Neuropathol (Berl) 33:129–138
Schmalbruch H (1982) The muscular dystrophies. In: Mastaglia FL, Walton JN (eds) Skeletal muscle pathology. Churchill-Livingstone, Edinburgh, pp 235–265
Sewry CA, Dubowitz V (1988) Histochemical and immunocytochemical studies in neuromuscular diseases. In: Walton JN (ed) Disorders of voluntary muscle, 5th edn. Churchill-Livingstone, Edinburgh, pp 241–283
Thornell L-E, Edstrom L, Eriksson A, Henriksson K-G, Angqvist K-A (1980) The distribution of intermediate filament protein (skeletin) in normal and diseased human skeletal muscle. J Neurol Sci 47:153–170
Tokuyasu KT (1980) Immunochemistry on ultrathin frozen sections. Histochem J 12:381–403
Tokuyasu KT (1986) Application of cryoultramicrotomy to immunocytochemistry. J Microsc 143:139–149
Tokuyasu KJ, Maher PA, Singer SJ (1984) Distributions of vimentin and desmin in developing chick myotubes in vivo. I. Immunofluorescence study. J Cell Biol 98:1961–1972
Tokuyasu KJ, Maher PA, Singer SJ (1985) Distributions of vimentin and desmin in developing chick myotubes in vivo. II. Immunoelectron microscopic study. J Cell Biol 100:1157–1166
Watkins SC, Samuel JL, Marotte F, Bertier-Savalle B, Rappaport L (1987) Microtubules and desmin filaments during onset of heart hypertrophy in rat. A double immunoelectron microscope study. Circ Res 60:327–336
Watkins SC, Hoffman EP, Slayter HS, Kunkel LM (1988) Immunoelectron microscopic localization of dystrophin in myofibres. Nature 333:863–866
Whiting A, Wardale J, Trinick J (1989) Does titin regulate the length of muscle thick filaments? J Mol Biol 205:263–268
Yoshioka T, Higuchi H, Kimura S, Ohashi K, Umazume Y, Maruyama K (1986) Effects of mild trypsin treatment on the passive tension generation and connectin splitting in stretched skinned fibers from frog skeletal muscle. Biomed Res 7:181–186
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Supported by the Muscular Dystrophy Group of Great Britain and Northern Ireland
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Cullen, M.J., Fulthorpe, J.J. & Harris, J.B. The distribution of desmin and titin in normal and dystrophic human muscle. Acta Neuropathol 83, 158–169 (1992). https://doi.org/10.1007/BF00308475
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DOI: https://doi.org/10.1007/BF00308475