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Passive stiffness of Drosophila IFM myofibrils: a novel, high accuracy

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Abstract

As the smallest muscle-cell substructure that retains the intact contractile apparatus, the single myofibril is considered the optimal specimen for muscle mechanics, although its small size also poses some technical difficulties. Myofibrils from Drosophila indirect flight muscle (IFM) are particularly difficult to study because their high passive stiffness makes them hard to handle, and too resistant to stretch to produce enough elongation for the accurate measurement of sarcomere length change. In this study, we devised a novel method for accurate stiffness measurement of single relaxed myofibrils using microfabricated cantilevers and phase contrast microscopy. A special experimental protocol was developed to minimize errors, and some data analysis strategies were used to identify and exclude spurious data. Remarkably consistent results were obtained from Drosophila IFM myofibrils. This novel, high accuracy method is potentially an effective tool for detecting small passive stiffness change in muscle mutants.

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References

  • Bartoo ML, Linke WA and Pollack GH (1997) Basis of passive tension and stiffness in isolated rabbit myofibrils. Am J Physiol 273: C266–276.

    PubMed  CAS  Google Scholar 

  • Blyakhman F, Tourovskaya A and Pollack GH (2001) Quantal sarcomere-length changes in relaxed single myofibrils. Biophys J 81: 1093–1100.

    Google Scholar 

  • Colomo F, Nencini S, Piroddi N, Poggesi C and Tesi C (1998) Calcium dependence of the apparent rate of force generation in single striated muscle myofibrils activated by rapid solution changes. Adv Exp Med Biol 453: 373–381.

    PubMed  CAS  Google Scholar 

  • Colomo F, Piroddi N, Poggesi C, te Kronnie G and Tesi C (1997) Active and passive forces of isolated myofibrils from cardiac and fast skeletal muscle of the frog. J Physiol 500 (Pt 2): 535–548.

    PubMed  CAS  Google Scholar 

  • Dickinson MH, Hyatt CJ, Lehmann FO, Moore JR, Reedy MC, Simcox A, Tohtong R, Vigoreaux JO, Yamashita H and Maughan DW (1997) Phosphorylation-dependent power output of transgenic flies: an integrated study. Biophys J 73: 3122–3134.

    PubMed  CAS  Google Scholar 

  • Fauver ME, Dunaway DL, Lilienfeld DH, Craighead HG and Pollack GH (1998) Microfabricated cantilevers for measurement of subcellular and molecular forces. IEEE Trans Biomed Eng 45: 891–898.

    Article  PubMed  CAS  Google Scholar 

  • Fearn LA, Bartoo ML, Myers JA and Pollack GH (1993) An optical fiber transducer for single myofibril force measurement. IEEE Trans Biomed Eng 40: 1127–1132.

    Article  PubMed  CAS  Google Scholar 

  • Friedman AL and Goldman YE (1996) Mechanical characterization of skeletal muscle myofibrils. Biophys J 71: 2774–2785.

    PubMed  CAS  Google Scholar 

  • Horowitz A and Pollack GH (1993) Force-length relation of isometric sarcomeres in fixed-end tetani. Am J Physiol 264: C19–26.

    PubMed  CAS  Google Scholar 

  • Irving T, Bhattacharya S, Tesic I, Moore J, Farman G, Simcox A, Vigoreaux J and Maughan D (2001) Changes in myofibrillar structure and function produced by N-terminal deletion of the regulatory light chain in Drosophila. J Muscle Res Cell Motil 22: 675–683.

    Article  PubMed  CAS  Google Scholar 

  • Irving TC and Maughan DW (2000) In vivo X-ray diffraction of indirect flight muscle from Drosophila melanogaster. Biophys J 78: 2511–2515.

    PubMed  CAS  Google Scholar 

  • Iwazumi T (1987) High-speed ultrasensitive instrumentation for myofibril mechanics measurements. Am J Physiol 252: C253–262.

    Google Scholar 

  • Kulke M, Neagoe C, Kolmerer B, Minajeva A, Hinssen H, Bullard B and Linke WA (2001) Kettin, a major source of myofibrillar stiffness in Drosophila indirect flight muscle. J Cell Biol 154: 1045–1057.

    Article  PubMed  CAS  Google Scholar 

  • Linke WA, Bartoo ML, Ivemeyer M and Pollack GH (1996) Limits of titin extension in single cardiac myofibrils. J Muscle Res Cell Motil 17: 425–438.

    Article  PubMed  CAS  Google Scholar 

  • Maughan DW and Vigoreaux JO (1999) An integrated view of insect flight muscle: genes, motor molecules, and motion. News Physiol Sci 14: 87–92.

    PubMed  CAS  Google Scholar 

  • Peckham M, Molloy JE, Sparrow JC and White DC (1990) Physiological properties of the dorsal longitudinal flight muscle and the tergal depressor of the trochanter muscle of Drosophila melanogaster. J Muscle Res Cell Motil 11: 203–215.

    Article  PubMed  CAS  Google Scholar 

  • Saide JD, Chin-Bow S, Hogan-Sheldon J, Busquets-Turner L, Vigoreaux JO, Valgeirsdottir K and Pardue ML (1989) Characterization of components of Z-bands in the fibrillar flight muscle of Drosophila melanogaster. J Cell Biol 109: 2157–2167.

    Article  PubMed  CAS  Google Scholar 

  • Tesi C, Colomo F, Nencini S, Piroddi N and Poggesi C (1999) Modulation by substrate concentration of maximal shortening velocity and isometric force in single myofibrils from frog and rabbit fast skeletal muscle. J Physiol 516 (Pt 3): 847–853.

    Article  PubMed  CAS  Google Scholar 

  • Tesi C, Piroddi N, Colomo F and Poggesi C (2002) Relaxation kinetics following sudden Ca(2+) reduction in single myofibrils from skeletal muscle. Biophys J 83: 2142–2151.

    Article  PubMed  CAS  Google Scholar 

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Hao, Y., Bernstein, S.I. & Pollack, G.H. Passive stiffness of Drosophila IFM myofibrils: a novel, high accuracy. J Muscle Res Cell Motil 25, 359–366 (2004). https://doi.org/10.1007/s10974-004-0684-5

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  • DOI: https://doi.org/10.1007/s10974-004-0684-5

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