Summary
The present study describes an experimental approach whereby myofibrillogenesis and the three-dimensional arrangement of myofibrils present within cultured skeletal muscle cells can be examined using the scanning electron microscope. This procedure uses cells that have been cultured on gold-coated coverslips, and treated with Triton X-100 to extract the cell membrane and the soluble cytoplasm. Subsequent electroconductive staining by treatment with thiocarbohydrazide and osmium allows the myofibrils to be visualized. The images of myofibrils in various states of development observed by this method generally accords to those previously reported by transmission electron microscopy. Cell elongation and adhesion to the substrate causes mechanical stress from different directions which meet at branchings of the cultured myotubes. Many myofibrils are observed to run in the direction of the inferred stress lines.
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References
Auber J (1969) La myofibrillogenèse du muscle strié. II. Vertébrés. J Microsc 8:367–390
Bell PB (1981) The application of scanning electron microscopy to the study of the cytoskeleton of cells in culture. Scanning Electron Microscopy/1981/II, 139–157
Bell PB, Miller MM, Carraway KL, Revel J-P (1978) SEM-revealed changes in the distribution of the Triton-insoluble cytoskeleton of Chinese hamster ovary cells induced by dibutyl cyclic AMP. Scanning Electron Microscopy/1978/II, 899–907
Boyd JD (1960) Development of striated muscle. In: Bourne GH (ed) The structure and function of muscle. 1st ed, Academic Press, New York, Vol 1, p 63
Brown S, Levinson W, Spudich JA (1976) Cytoskeletal elements of chick embryo fibroblasts revealed by detergent extraction. J Supramol Struct 5:119–130
Cantini M, Sartore S, Schiaffino S (1980) Myosin types in cultured muscle cells. J Cell Biol 85:903–909
Carey EJ (1920) Studies in the dynamics of histogenesis. I. Tension of differential growth as a stimulus to myogenesis. J Gen Physiol 2:357–372
Fischman DA (1967) An electron microscope study of myofibril formation in embryonic chick skeletal muscle. J Cell Biol 32:557–575
Fischman DA (1970) The synthesis and assembly of myofibrils in embryonic muscle. Curr Top Dev Biol 5:235–280
Fischman DA (1972) Development of striated muscle. In Bourne GH (ed) The structure and function of muscle. 2nd ed, Academic Press, New York, Vol 1, p 75
Fulton AB, Prives J, Farmer SR, Penman S (1981) Developmental reorganization of the skeletal framework and its surface lamina in fusing muscle cells. J Cell Biol 91:103–112
Henderson D, Weber K (1979) Three dimensional organization of microfilaments and microtubules in the cytoskeleton. Exp Cell Res 124:301–316
Heuser JE, Kirschner MW (1980) Filament organization revealed in platinum replicas of freezedried cytoskeletons. J Cell Biol 86:212–234
Heuson-Stienon JA (1965) Morphogenèse de la cellule musculaire striée au microscope électronique. J Microsc 4:657–678
Ip W, Fischman DA (1979) High resolution scanning electron microscopy of isolated and in situ cytoskeletal elements. J Cell Biol 83:249–254
Kelley RO, Dekker RA, Bluemink JG (1973) Ligand-mediated osmium binding: its application in coating biological specimens for scanning electron microscopy. J Ultrastruct Res 45:254–258
Kelly DE (1969) Myofibrillogenesis and Z-band differentiation. Anat Rec 163:403–426
Kulikowski RR (1981) Myofibrillogenesis in vitro: implications for early cardiac morphogenesis. In: Pexieder T (ed) Perspectives in cardiovascular research. Vol 5, Mechanisms of cardiac morphogenesis and teratogenesis. Academic Press, New York, p 367
Laemmli UK (1970) Cleavage of structural proteins during the assembly of head of bacteriophage T4. Nature (London) 227:680–685
Osborn M, Weber K (1977) The detergent-resistant cytoskeleton of tissue culture cells includes the nucleus and the microfilament bundles. Exp Cell Res 106:339–349
Peng HB, Wolosewick JJ, Cheng P-C (1981) The development of myofibrils in cultured muscle cells: a whole-mount and thin-section electron microscopic study. Dev Biol 88:121–136
Porter KR, Anderson K (1980) The morphogenesis of myofibrils from the microtrabecular lattice as observed in cultured myoblasts. In: Ebashi S, Maruyama K, Endo M (eds) Muscle contraction: its regulatory mechanism. Japan Sci Soc Press/Springer Tokyo/Berlin Heidelberg New York, p 527
Pudney J, Singer RH (1979) Electron microscopic visualization of the filamentous reticulum in whole cultured presumptive chick myoblasts. Am J Anat 156:321–326
Richler C, Yaffe D (1970) The in vitro cultivation and differentiation capacities of myogenic cell lines. Dev Biol 23:1–22
Rubinstein NA, Holtzer H (1979) Fast and slow muscles in tissue culture synthesize only fast myosin. Nature (London) 280:323–325
Schliwa M, van Blerkom J (1981) Structural interaction of cytoskeletal components. J Cell Biol 90:222–235
Shimada Y, Fischman DA, Moscona AA (1967) The fine structure of embryonic chick skeletal muscle cells differentiated in vitro. J Cell Biol 35:445–453
Thornell L-E, Sjöström M, Andersson K-E (1976) The relationship between mechanical stress and myofibrillar organization in heart Purkinje fibers. J Mol Cell Card 8:689–695
Trotter JA, Beverly A, Foeder BA, Keller JM (1978) Intracellular fibers in cultured cells: analysis by scanning and transmission electron microscopy and by SDS-polyacrylamide gel electrophoresis. J Cell Sci 31:369–392
Warren RH (1973) Interaction of the sarcoplasmic reticulum with Z-lines during myogenesis in amphibian skeletal muscle. Anat Rec 177:225–242
Webster RE, Henderson D, Osborn M, Weber K (1978) Three-dimensional electron microscopical visualization of the cytoskeleton of animal cells: immunoferritin identification of actinand tubulin-containing structures. Proc Natl Acad Sci USA 75:5511–5515
Yaffe D (1968) Retention of differentiation potentialities during prolonged cultivation of myogenic cells. Proc Natl Acad Sci USA 61:477–483
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Isobe, Y., Shimada, Y. Myofibrillogenesis in vitro as seen with the scanning electron microscope. Cell Tissue Res. 231, 481–494 (1983). https://doi.org/10.1007/BF00218107
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DOI: https://doi.org/10.1007/BF00218107