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Cell and Tissue Research

, Volume 263, Issue 3, pp 419–430 | Cite as

Chicken cardiac myofibrillogenesis studied with antibodies specific for titin and the muscle and nonmuscle isoforms of actin and tropomyosin

  • Susan E. Handel
  • Marion L. Greaser
  • Edward Schultz
  • Seu-Mei Wang
  • Jeannette C. Bulinski
  • Jim J. -C. Lin
  • James L. Lessard
Article

Summary

Myofibrillogenesis was studied in cultured chick cardiomyocytes using indirect immunofluorescence microscopy and antibodies against α- and γ-actin, muscle and nonmuscle tropomyosin, muscle myosin, and titin. Initially, cardiomyocytes, devoid of myofibrils, developed variable numbers of stress fiber-like structures with uniform staining for anti-muscle and nonmuscle actin and tropomyosin, and diffuse, weak staining with anti-titin. Anti-myosin labeled bundles of filaments that exhibited variable degrees of association with the stress fiber-like structures. Myofibrillogenesis occurred with a progressive, and generally simultaneous, longitudinal reorganization of stress fiber-like structures to form primitive sarcomeric units. Titin appeared to attain its mature pattern before the other major contractile proteins. Changes in the staining patterns of actin, tropomyosin, and myosin as myofibrils matured were interpreted as due to longitudinal filament alignment occurring before ordering in the axial direction. Non-muscle actin and tropomyosin were found with sarcomeric periodicity in the initial stages of sarcomere myofibrillogenesis, although their staining patterns were not identical. The localization of the “sarcomeric” proteins α-actin and muscle tropomyosin in stress fiber-like structures and the incorporation of non-muscle proteins in the initial stages of sarcomere organization bring into question the meaning of “sarcomeric” proteins in regard to myofibrillogenesis.

Key words

Fibrillogenesis Muscle, cardiac Myosin Actin Immunofluorescence microscopy Myofibrils Domestic fowl 

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References

  1. Antin PB, Tokunaka S, Nachmias VT, Holtzer H (1986) Role of stress fiber-like structures in assembling nascent myofibrils in myosheets recovering from exposure to ethyl methanesulfonate. J Cell Biol 102:1464–1479Google Scholar
  2. Bader D, Masaki T, Fischman DA (1982) Immunochemical analysis of myosin heavy chain during avian myogenesis in vivo and in vitro. J Cell Biol 95:763–770Google Scholar
  3. Bulinski JC, Kumar S, Titani K, Hauschka SD (1983) Peptide antibody specific for the amino terminus of skeletal muscle α-actin. Proc Natl Acad Sci USA 80:1506–1510Google Scholar
  4. Dlugosz AA, Antin PB, Nachmias VT, Holtzer H (1984) The relationship between stress fiber-like structures and nascent myofibrils in cultured cardiac myocytes. J Cell Biol 99:2268–2278Google Scholar
  5. Fallon JR, Nachmias VT (1980) Localization of cytoplasmic and skeletal myosins in developing muscle cells by double-label immunofluorescence. J Cell Biol 87:237–247Google Scholar
  6. Furst 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 109:517–527Google Scholar
  7. Glacy SD (1982) Subcellular distribution of rhodamine-actin microinjected in living fibroblastic cells. J Cell Biol 97:1207–1213Google Scholar
  8. Greaser ML, Handel SE, Wang S-M, Schultz E, Bulinski JC, Lin JJ-C, Lessard JL (1989) Assembly of titin, myosin, actin, and tropomyosin into myofibrils in cultured chick cardiomyocytes. In: Stockdale F, Kedes L (eds) Cellular and molecular biology of muscle development. UCLA Symposia on Molecular and Cellular Biology; New Series; vol 93. Liss, New York, pp 247–257Google Scholar
  9. Handel SE, Greaser ML, Wang S-M, Schultz E, Bulinski JC, Lin JJ-C, Lessard JL (1986) Myofibrillogenesis in chick cardiac myocytes as studied with antibodies against muscle and nonmuscle proteins. J Cell Biol 103:125aGoogle Scholar
  10. 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 103:2185–2196Google Scholar
  11. Kaehn K, Bachmann P, Falkenberg FW (1985) Immunofluorescence staining of thin-filament sections not participating in actomyosin cross-bridges: studies by use of a monoclonal antibody specific to actin. Cell Tissue Res 239:417–422Google Scholar
  12. Lessard JL (1988) Two monoclonal antibodies to actin: one muscle selective and one general reactive. J Cell Motil Cytoskel 10:349–362Google Scholar
  13. Lessard JL, Scheffter S, Engel L, Tepperman K (1983) Immunofluorescent localization of actins in differentiating chick myoblasts. J Cell Biol 97:74aGoogle Scholar
  14. Lin JJ-C, Lin JL-C (1986) Assembly of different isoforms of actin and tropomyosin into the skeletal tropomyosin-enriched microfilaments during differentiation of muscle cells in vitro. J Cell Biol 103:2173–2183Google Scholar
  15. Lin JJ-C, Chou C-S, Lin JL-C (1985) Monoclonal antibodies against chicken tropomyosin isoforms: production, characterization and application. Hybridoma 4:223–242Google Scholar
  16. Lin Z, Eshelman JR, Forry-Schaudies S, Duran S, Lessard JL, Holtzer H (1987) Sequential disassembly of myofibrils induced by myristate acetate in cultured myotubes. J Cell Biol 105:1365–1376Google Scholar
  17. Lin Z, Holtzer S, Schultheiss T, Murray J, Masaki T, Fischman DA, Holtzer H (1989) Polygons and adhesion plaques and the disassembly and assembly of myofibrils in cardiac myocytes. J Cell Biol 108:2355–2367Google Scholar
  18. McKenna NM, Meigs JB, Wang Y-L (1985) Identical distribution of fluorescently labeled brain and muscle actins in living cardiac fibroblasts and myocytes. J Cell Biol 100:292–296Google Scholar
  19. Otey CA, Kalnoski MH, Lessard JL, Bulinski JC (1986) Immunolocalization of the gamma isoform of non-muscle actin in cultured cells. J Cell Biol 102:1726–1737Google Scholar
  20. Otey CA, Kalnoski MH, Bulinski JC (1988) Immunolocalization of muscle and non-muscle isoforms of actin in myogenic cells and adult skeletal muscle. J Cytoskel Cell Motil 9:337–348Google Scholar
  21. Peng HB, Wolosewick JJ, Cheng P-C (1981) The development of myofibrils in cultured muscle cells: a whole-mount and thinsection electron microscopic study. Dev Biol 88:121–136Google Scholar
  22. Sanger JW (1977) Mitosis in beating cardiac myoblasts treated with cytochalasin-B. J Exp Zool 201:403–409Google Scholar
  23. Sanger JM, Mittal B, Pochapin MB, Sanger JW (1986) Myofibrillogenesis in living cells microinjected with fluorescently labeled alpha-actinin. J Cell Biol 102:2053–2066Google Scholar
  24. Schultheiss T, Lin Z, Lu M-H, Murray J, Fischman D, Weber K, Masaki T, Imamura M, Holtzer H (1990) Differential distribution of subsets of myofibrillar proteins in cardiac nonstriated and striated myofibrils. J Cell Biol 110:1159–1172Google Scholar
  25. Terai M, Komiyama M, Shimada Y (1989) Myofibril assembly is linked with vinculin, α-actinin, and cell-substrate contacts in embryonic cardiac myocytes in vitro. Cell Motil Cytoskel 12:185–194Google Scholar
  26. Tokuyasu KT (1989) Immunocytochemical studies of cardiac myofibrillogenesis in early chick embryos. III. Generation of fasciae adherentes and costameres. J Cell Biol 108:43–53Google Scholar
  27. Tokuyasu KT, Maher PA (1987) Immunochemical studies of cardiac myofibrillogenesis in early chick embryos. I. Presence of immunofluorescent titin spots in premyofibril stages. J Cell Biol 105:2781–2793Google Scholar
  28. Wang S-M, Greaser ML (1985) Immunocytochemical studies using a monoclonal antibody to bovine cardiac titin on intact and extracted myofibrils. J Muscle Res Cell Motil 6:293–312Google Scholar
  29. Wang S-M, Greaser ML, Schultz E, Bulinski JC, Lin JJ-C, Lessard JL (1988) Studies on cardiac myofibrillogenesis with antibodies to titin, actin, tropomyosin and myosin. J Cell Biol 107:1075–1083Google Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • Susan E. Handel
    • 1
  • Marion L. Greaser
    • 1
    • 2
  • Edward Schultz
    • 2
  • Seu-Mei Wang
    • 1
    • 2
  • Jeannette C. Bulinski
    • 3
  • Jim J. -C. Lin
    • 4
  • James L. Lessard
    • 5
  1. 1.Muscle Biology LaboratoryUniversity of Wisconsin-MadisonMadisonUSA
  2. 2.Department of AnatomyUniversity of Wisconsin-MadisonMadisonUSA
  3. 3.Department of Anatomy and Cell BiologyColumbia UniversityNew YorkUSA
  4. 4.Department of BiologyUniversity of IowaIowa CityUSA
  5. 5.Division of Basic ResearchChildrens Hospital Research FoundationCincinnatiUSA

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