Whole-Cell Voltage Clamp on Skeletal Muscle Fibers With the Silicone-Clamp Technique

  • Sandrine Pouvreau
  • Claude Collet
  • Bruno Allard
  • Vincent Jacquemond
Part of the Methods in Molecular Biology™ book series (MIMB, volume 403)


Control of membrane voltage and membrane current measurements are of strong interest for the study of numerous aspects of skeletal muscle physiology and pathophysiology. The silicone-clamp technique makes use of a conventional patch-clamp apparatus to achieve whole-cell voltage clamp of a restricted portion of a fully differentiated adult skeletal muscle fiber. The major part of an isolated muscle fiber is insulated from the extracellular medium with silicone grease, and the tip of a single microelectrode connected to the amplifier is then inserted within the fiber through the silicone layer. This method represents an alternative to the traditional vaseline-gap isolation and two or three microelectrode voltage-clamp techniques. This chapter reviews the main benefits of the silicone-clamp technique and provides detailed insights into its practical implementation.

Key Words

Skeletal muscle voltage clamp silicone grease mammalian muscle ion channels excitation–contraction coupling 



This work was supported by the Centre National de la Recherche Scientifique, the Université Claude Bernard Lyon 1, and the Association Française contre les Myopathies.


  1. 1.
    Adrian, R. H., Chandler, W. K., and Hodgkin, A. L. (1966) Voltage clamp experiments in skeletal muscle fibres. J. Physiol. 186, 51P–52P.PubMedGoogle Scholar
  2. 2.
    Adrian, R. H., Chandler, W. K., and Hodgkin, A. L. (1970) Voltage clamp experiments in striated muscle fibres. J. Physiol. 208, 607–644.PubMedGoogle Scholar
  3. 3.
    Chandler, W. K., Rakowski, R. F., and Schneider, M. F. (1976) A non-linear voltage dependent charge movement in frog skeletal muscle. J. Physiol. 254, 245–283.PubMedGoogle Scholar
  4. 4.
    Heistracher, P. and Hunt, C. C. (1969) The relation of membrane changes to contraction in twitch muscle fibres. J. Physiol. 201, 589–611.PubMedGoogle Scholar
  5. 5.
    Adrian, R. H. and Marshall, M. W. (1977) Sodium currents in mammalian muscle. J. Physiol. 268, 223–250.PubMedGoogle Scholar
  6. 6.
    Ildefonse, M. and Rougier, O. (1972) Voltage-clamp analysis of the early current in frog skeletal muscle fibre using the double sucrose-gap method. J. Physiol. 222, 373–395.PubMedGoogle Scholar
  7. 7.
    Hille, B. and Campbell, D. T. (1976) An improved vaseline gap voltage clamp for skeletal muscle fibers. J. Gen. Physiol. 67, 265–293.PubMedCrossRefGoogle Scholar
  8. 8.
    Kovacs, L. and Schneider, M. F. (1978) Contractile activation by voltage clamp depolarization of cut skeletal muscle fibres. J. Physiol. 277, 483–506.PubMedGoogle Scholar
  9. 9.
    Kovacs, L., Rios, E., and Schneider, M. F. (1983) Measurement and modification of free calcium transients in frog skeletal muscle fibres by a metallochromic indicator dye. J. Physiol. 343, 161–196.PubMedGoogle Scholar
  10. 10.
    Bekoff, A. and Betz, W. J. (1977) Physiological properties of dissociated muscle fibres obtained from innervated and denervated adult rat muscle. J. Physiol. 271, 25–40.PubMedGoogle Scholar
  11. 11.
    Szentesi, P., Jacquemond, V., Kovacs, L., and Csernoch, L. (1997) Intramembrane charge movement and sarcoplasmic calcium release in enzymatically isolated mammalian skeletal muscle fibres. J. Physiol. 505, 371–384.PubMedCrossRefGoogle Scholar
  12. 12.
    Woods, C. E., Novo, D., DiFranco, M., and Vergara, J. L. (2004) The action potential-evoked sarcoplasmic reticulum calcium release is impaired in mdx mouse muscle fibres. J. Physiol. 557, 59–75.PubMedCrossRefGoogle Scholar
  13. 13.
    Jacquemond, V. (1997) Indo-1 fluorescence signals elicited by membrane depolarization in enzymatically isolated mouse skeletal muscle fibers. Biophys. J. 73, 920–928.PubMedCrossRefGoogle Scholar
  14. 14.
    Jacquemond, V. and Allard, B. (1998). Activation of Ca2+-activated K+ channels by an increase in intracellular Ca2+ induced by depolarization of mouse skeletal muscle fibres. J. Physiol. 509, 93–102.PubMedCrossRefGoogle Scholar
  15. 15.
    Collet, C., Pouvreau, S., Csernoch, L., Allard, B., and Jacquemond, V. (2004) Calcium signaling in isolated skeletal muscle fibers investigated under “silicone voltage-clamp” conditions. Cell Biochem. Biophys. 40, 225–236.PubMedCrossRefGoogle Scholar
  16. 16.
    Csernoch, L., Bernengo, J. C., Szentesi, P., and Jacquemond, V. (1998) Measurements of intracellular Mg2+ concentration in mouse skeletal muscle fibers with the fluorescent indicator mag-indo-1. Biophys. J. 75, 957–967.PubMedCrossRefGoogle Scholar
  17. 17.
    Bernengo, J. C., Collet, C., and Jacquemond, V. (2001) Intracellular Mg2+ diffusion within isolated rat skeletal muscle fibers. Biophys. Chem. 89, 35–51.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2007

Authors and Affiliations

  • Sandrine Pouvreau
  • Claude Collet
  • Bruno Allard
  • Vincent Jacquemond

There are no affiliations available

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