Summary
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.
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References
Adrian, R. H., Chandler, W. K., and Hodgkin, A. L. (1966) Voltage clamp experiments in skeletal muscle fibres. J. Physiol. 186, 51P–52P.
Adrian, R. H., Chandler, W. K., and Hodgkin, A. L. (1970) Voltage clamp experiments in striated muscle fibres. J. Physiol. 208, 607–644.
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.
Heistracher, P. and Hunt, C. C. (1969) The relation of membrane changes to contraction in twitch muscle fibres. J. Physiol. 201, 589–611.
Adrian, R. H. and Marshall, M. W. (1977) Sodium currents in mammalian muscle. J. Physiol. 268, 223–250.
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.
Hille, B. and Campbell, D. T. (1976) An improved vaseline gap voltage clamp for skeletal muscle fibers. J. Gen. Physiol. 67, 265–293.
Kovacs, L. and Schneider, M. F. (1978) Contractile activation by voltage clamp depolarization of cut skeletal muscle fibres. J. Physiol. 277, 483–506.
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.
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.
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.
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.
Jacquemond, V. (1997) Indo-1 fluorescence signals elicited by membrane depolarization in enzymatically isolated mouse skeletal muscle fibers. Biophys. J. 73, 920–928.
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.
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.
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.
Bernengo, J. C., Collet, C., and Jacquemond, V. (2001) Intracellular Mg2+ diffusion within isolated rat skeletal muscle fibers. Biophys. Chem. 89, 35–51.
Acknowledgments
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.
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Pouvreau, S., Collet, C., Allard, B., Jacquemond, V. (2007). Whole-Cell Voltage Clamp on Skeletal Muscle Fibers With the Silicone-Clamp Technique. In: Molnar, P., Hickman, J.J. (eds) Patch-Clamp Methods and Protocols. Methods in Molecular Biology™, vol 403. Humana Press. https://doi.org/10.1007/978-1-59745-529-9_12
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DOI: https://doi.org/10.1007/978-1-59745-529-9_12
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