PubMed 25023317

Referenced in Channelpedia wiki pages of: none

Automatically associated channels: TRP , TRPM , TRPM4

Title: Current recordings at the single channel level in adult mammalian isolated cardiomyocytes.

Authors: Romain Guinamard, Thomas Hof, Laurent Sallé

Journal, date & volume: Methods Mol. Biol., 2014 , 1183, 291-307

PubMed link:

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).