Analysis of calcium transients in cardiac myocytes and assessment of the sarcoplasmic reticulum Ca2+-ATPase contribution.

Ca(2+) signaling plays an essential role in several functions of cardiac myocytes. Transient rises and reductions of cytosolic Ca(2+), permitted by the sarcoplasmic reticulum Ca(2+) ATPase (SERCA2) and other proteins, control each cycle of contraction and relaxation. Here we provide a practical method for isolation of neonatal rat cardiac myocytes and measurement of Ca(2+) transients in cultured cardiac myocytes, yielding information on kinetic resolution of the transients, variations of cytosolic Ca(2+) concentrations, and adequacy of intracellular Ca(2+) stores. We also provide examples of experimental perturbations that can be used to assess the contribution of SERCA2 to Ca(2+) signaling.