Analysis of Calcium Transients in Cardiac Myocytes and Assessment of the Sarcoplasmic Reticulum Ca2+-ATPase Contribution

Ca²⁺ signaling plays an essential role in several functions of cardiac myocytes. Transient rises and reductions of cytosolic Ca²⁺ , permitted by the sarcoplasmic reticulum Ca²⁺ 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 Ca²⁺ transients in cultured cardiac myocytes, yielding information on kinetic resolution of the transients, variations of cytosolic Ca²⁺ concentrations, and adequacy of intracellular Ca²⁺ stores. We also provide examples of experimental perturbations that can be used to assess the contribution of SERCA2 to Ca²⁺ signaling.