Pflügers Archiv

, Volume 417, Issue 3, pp 303–308 | Cite as

Calcium sensitivity and myofibrillar protein isoforms of rat skinned skeletal muscle fibres

  • D. Danieli -Betto
  • R. Betto
  • M. Midrio
Excitable Tissues and Central Nervous Physiology
Received:
Revised:
Accepted:

Abstract

We investigated the calcium sensitivity for tension generation of different fibre types and the possible correlation between calcium sensitivity and the presence of distinct regulatory protein and myosin light chain (MLC) isoforms in rat skinned skeletal muscle fibres. Fibre types 1, 2A and 2B were identified by electrophoretic analysis of myosin heavy chain (MHC) isoforms. Fibres showing more than one MHC isoform were discarded. Type 1 fibres from the soleus showed a higher pCa (−log10 [Ca], where [ ] denotes concentration) threshold and a lower slope of pCa/tension curve than type 2 extensor digitorum longus (EDL) fibres; between type 2 fibres, type 2B showed the higher slope of pCa/tension curve. Type 1 fibres from different muscles showed similar calcium sensitivities when containing only the slow set of regulatory proteins and MLC; when both slow and fast isoforms were present, calcium sensitivity shifted toward fast type fibre values. Type 2A fibres from different muscles showed a similar calcium sensitivity, independently of the set (purely fast or mixed) of regulatory proteins and MLC. It is suggested that when both fast and slow isoforms of regulatory proteins and of MLC are present in a muscle fibre, calcium sensitivity is dictated mainly by the fast isoforms.

Key words

Skinned fibres Calcium sensitivity Muscle fibre types 
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References

  1. Babu A, Scordilis SP, Sonnenblick EH, Gulati J (1987) The control of myocardial contraction with skeletal fast muscle troponin C. J Biol Chem 262:5815–5822Google Scholar
  2. Brandt PW, Diamond MS, Gluck B, Kaway M, Schachat F (1984) Molecular basis of cooperativity in vertebrate muscle thin filaments. Carlsberg Res Commun 49:155–167Google Scholar
  3. Danieli-Betto D, Zerbato E, Betto R (1986) Type 1, 2A and 2B myosin heavy chain electrophoretic analysis of rat muscle fibers. Biochem Biophys Res Commun 138:981–987Google Scholar
  4. Donaldson SKB (1984) Ca2+-activated force-generting properties of mammalian skeletal muscle fibres: histochemically identified single peeled rabbit fibres. J Mus Res Cell Motil 5:593–612Google Scholar
  5. Dulhunty AF, Gage PW (1983) Asymmetrical charge movement in slow- and fast-twitch mammalian muscle fibres in normal and paraplegic rats. J Physiol (Lond) 341:213–231Google Scholar
  6. Eddinger TJ, Moss RL (1987) Mechanical properties of skinned single fibers of identified types from rat diaphragm. Am J Physiol 253:C210-C218Google Scholar
  7. Gordon AM, Ridway EB, Yates LD, Allen T (1988) Muscle crossbridge attachment: effects on calcium binding and calcium activation. In: Sugi H, Pollack GH (eds) Molecular mechanism of muscle contraction. Plenum Press, New York, pp 89–99Google Scholar
  8. Grabarek Z, Grabarek J, Leavis PC, Gergely J (1983) Cooperative binding to the Ca2+-specific sites of troponin C in regulated actin and actomyosin. J Biol Chem 258:14098–14102Google Scholar
  9. Grabarek Z, Leavis PC, Gergely J (1986) Calcium binding to the low affinity sites in troponin C induces conformational changes in the high affinity domain. J Biol Chem 261:608–613Google Scholar
  10. Greaser ML, Moss RL, Reiser PJ (1988) Variations in contractile properties of rabbit single muscle fibres in relation to troponin T isoforms and myosin light chains. J Physiol (Lond) 406:85–98Google Scholar
  11. Gulati J, Scordilis SP, Babu A (1988) Effect of troponin C on the cooperativity in Ca2+ activation of cardiac muscle. FEBS Lett 236:441–444Google Scholar
  12. Hofmann PA, Metzger JM, Greaser ML, Moss RL (1990) Effects of partial extraction of light chain 2 on the Ca sensitivity of isometric tension, stiffness, and velocity of shortening in skinned skeletal muscle fibers. J Gen Physiol 95:477–498Google Scholar
  13. Kerrick WGL, Secrist D, Coby R, Lucas S (1976) Development of differences between red and white muscles in sensitivity to Ca2+ in the rabbit from embryo to adult. Nature 260:440–442Google Scholar
  14. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685Google Scholar
  15. Laszewski-Williams B, Ruff RL, Gordon AM (1989) Influence of fiber type and muscle source on Ca2+ sensitivity of rat fibers. Am J Physiol 256:C420-C427Google Scholar
  16. Metzger JM, Greaser ML, Moss RL (1989) Variations in chrossbridge attachment rate and tension with phosphorylation of myosin in mammalian skinned skeletal muscle fibers. Implication for twitch potentiation in intact muscle. J Gen Physiol 93:855–883Google Scholar
  17. Morano I, Arndt H, Gartner C, Ruegg JC (1988) Skinned fibers of human atrium and ventricle: Myosin isoenzymes and contractility. Circ Res 62:632–639Google Scholar
  18. Moss RL, Swinford AE, Greaser ML (1983) Alteration in the Ca2+ sensitivity of tension development by single skeletal muscle fibers at stretched lengths. Biophys J 43:115–119Google Scholar
  19. Moss RL, Lauer MR, Giulian GG, Greaser ML (1986) Altered Ca2+ dependence of tension development in skinned skeletal muscle fibers following modification of troponin by partial substitution with cardiac troponin C. J Biol Chem 261:6096–6099Google Scholar
  20. Mounier Y, Holy X, Stevens L (1989) Compared properties of the contractile system of skinned slow and fast rat muscle fibres. Pflügers Arch 415:136–141Google Scholar
  21. Orentlicher M, Brandt PW, Reuben JP (1977) Regulation of tension in skinned muscle fibers: effect of high concentration of Mg-ATP. Am J Physiol 233:C127-C134Google Scholar
  22. Reiser PJ, Greaser ML, Moss RL (1987) Tension/pCa characteristics and regulatory proteins of single fibers from chicken neonatal and adult fast and slow skeletal muscles. Biophys J 51:222aGoogle Scholar
  23. Ruegg JC (1986) Calcium in muscle activation. A comparative approach. Springer, Berlin Heidelberg New YorkGoogle Scholar
  24. Ruff RJ (1989) Calcium-sensitivity of fast- and slow-twitch human muscle fibers. Muscle Nerve 12:32–37Google Scholar
  25. Salviati G, Betto R, Danieli-Betto D (1982) Polymorphism of myofibrillar proteins of rabbit skeletal-muscle fibres. An electrophoretic study of single fibres. Biochem J 207:261–272Google Scholar
  26. Schachat FH, Diamond MS, Brandt PW (1987) Effect of different troponin T-tropomyosin combination on thin filament activation. J Mol Biol 198:551–554Google Scholar
  27. Staron RS, Pette D (1987a) The multiplicity of combinations of myosin light chains and heavy chains in histochemically typed single fibres. Rabbit soleus muscle. Biochem J 243:687–693Google Scholar
  28. Staron RS, Pette D (1987b) The multiplicity of combinations of myosin light chains and heavy chains in histochemically typed single fibres. Rabbit tibialis anterior muscle. Biochem J 243:695–699Google Scholar
  29. Stephenson DG, Forrest QG (1980) Different isometric force-[Ca2+] relationships in slow and fast twitch skinned muscle fibres of the rat. Biochim Biophys Acta 589:358–362Google Scholar
  30. Stephenson DG, Williams DA (1981) Calcium-activated force responses in fast- and slow-twitch skinned muscle fibres of the rat at different temperatures. J Physiol (Lond) 317:281–302Google Scholar
  31. Stephenson DG, Williams DA (1982) Effects of sarcomere length on the force-pCa relation in fast- and slow-twitch skinned fibres from the rat. J Physiol (Lond) 333:637–653Google Scholar
  32. Sweeney HL, Stull JT (1986) Phosphorylation of myosin in permeabilized mammalian cardiac and skeletal muscle cells. Am J Physiol 250:C657-C660Google Scholar
  33. Sweeney HL, Stull JT (1990) Alteration of cross-bridge kinetics by myosin light chain phosphorylation in rabbit skeletal muscle: implications for regulation of actin-myosin interaction. Proc Natl Acad Sci USA 87:414–418Google Scholar
  34. Takagi A, Endo M (1977) Guiena pig soleus and extensor digitorum longus: a study of single-skinned fibers. Exp Neurol 55:95–101Google Scholar
  35. Westwood SA, Hudlicka O, Perry SV (1984) Phosphorylation in vivo of the P light chain of myosin in rabbit fast and slow skeletal muscles. Biochem J 218:841–847Google Scholar
  36. Zeman RJ, Wood DS (1980) Correlative histochemical and physiological measurements in single skinned muscle fibers: heterogeneity of Ca sensitivity in type I fibers. J Histochem Cytochem 28:714–715Google Scholar

Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • D. Danieli -Betto
    • 1
  • R. Betto
    • 2
  • M. Midrio
    • 1
  1. 1.Istituto di Fisiologia UmanaUniversità di PadovaPaduaItaly
  2. 2.Consiglio Nazionale delle RicercheCentro di Studio per la Biologia e la Fisiopatologia MuscolarePaduaItaly

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