Intracellular Ca2+release underlies the development of phase 2 in mouse ventricular action potentials

Abstract

The ventricular action potential (AP) is characterized by a fast depolarizing phase followed by a repolarization that displays a second upstroke known as phase 2. This phase is generally not present in mouse ventricular myocytes. Thus we performed colocalized electrophysiological and optical recordings of APs in Langendorff-perfused mouse hearts founding a noticeable phase 2. Ryanodine as well as nifedipine reduced phase 2. Our hypothesis is that a depolarizing current activated by Ca2+released from the sarcoplasmic reticulum (SR) rather than the “electrogenicity” of the L-type Ca2+current is crucial in the generation of mouse ventricular phase 2. When Na+was partially replaced by Li+in the extracellular perfusate or the organ was cooled down, phase 2 was reduced. These results suggest that the Na+/Ca2+exchanger functioning in the forward mode is driving the depolarizing current that defines phase 2. Phase 2 appears to be an intrinsic characteristic of single isolated myocytes and not an emergent property of the tissue. As in whole heart experiments, ventricular myocytes impaled with microelectrodes displayed a large phase 2 that significantly increases when temperature was raised from 22 to 37°C. We conclude that mouse ventricular APs display a phase 2; however, changes in Ca2+dynamics and thermodynamic parameters also diminish phase 2, mostly by impairing the Na+/Ca2+exchanger. In summary, these results provide important insights about the role of Ca2+release in AP ventricular repolarization under physiological and pathological conditions.