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Current Recordings at the Single Channel Level in Adult Mammalian Isolated Cardiomyocytes

  • Romain GuinamardEmail author
  • Thomas Hof
  • Laurent Sallé
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1183)

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).

Key words

Single channel Patch clamp Cell isolation Mammalian cardiomyocytes Langendorff Calcium-activated non-selective cation channel TRPM4 

Notes

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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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Groupe Signalisation, Electrophysiologie et Imagerie des Lésions d’Ischémie/Reperfusion Myocardique, EA4650Normandie UniversitéCaen CedexFrance

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