Abstract
This chapter describes appropriate methods to investigate mammalian cardiac channels properties at the single channel level. Cell isolation is performed from new born or adult heart by enzymatic digestion on minced tissue or using the Langendorff apparatus. Isolation proceeding is suitable for rabbit, rat, and mouse hearts. In addition, isolation of human atrial cardiomyocytes is described. Such freshly isolated cells or cells maintained in primary culture are suitable for patch-clamp studies. Here we describe the single channel variants of the patch-clamp technique (cell-attached, inside-out, outside-out) used to investigate channel properties. Proceedings for the evaluation of biophysical properties such as conductance, ionic selectivity, regulations by extracellular and intracellular mechanisms are described. To illustrate the study, we provide an example by the characterization of a calcium-activated non-selective cation channel (TRPM4).
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Acknowledgments
The author is in indebted to Pr. Patrick Bois and Pr. Jacques Teulon for helpful comments on this manuscript and teaching in both tissue preparation and patch-clamp proceedings.
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Guinamard, R., Hof, T., Sallé, L. (2014). Current Recordings at the Single Channel Level in Adult Mammalian Isolated Cardiomyocytes. In: Martina, M., Taverna, S. (eds) Patch-Clamp Methods and Protocols. Methods in Molecular Biology, vol 1183. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1096-0_19
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DOI: https://doi.org/10.1007/978-1-4939-1096-0_19
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