title: Current recordings at the single channel level in adult mammalian isolated cardiomyocytes.
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).
authors.affiliation: Groupe Signalisation, Electrophysiologie et Imagerie des Lésions d'Ischémie/Reperfusion Myocardique, EA4650, Normandie Université, Esplanade de la Paix, 14032, Caen Cedex, France, romain.guinamard@unicaen.fr.
authors.forename: Romain
authors.initials: R
authors.lastname: Guinamard
authors.forename: Thomas
authors.initials: T
authors.lastname: Hof
authors.forename: Laurent
authors.initials: L
authors.lastname: Sallé
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D002478:Cells, Cultured; D004867:Equipment Design; D006325:Heart Atria / Q000166:cytology; D006352:Heart Ventricles / Q000166:cytology; D006801:Humans; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism*; D018408:Patch-Clamp Techniques / Q000379:methods*; D050053:TRPM Cation Channels / Q000378:metabolism*
chemical_list: D050053:TRPM Cation Channels