Phase 1 repolarization rate defines Ca2+ dynamics and contractility on intact mouse hearts

Abstract

In the heart, Ca²⁺ influx through L-type Ca²⁺ channels triggers Ca²⁺ release from the sarcoplasmic reticulum. In most mammals, this influx occurs during the ventricular action potential (AP) plateau phase 2. However, in murine models, the influx through L-type Ca²⁺ channels happens in early repolarizing phase 1. The aim of this work is to assess if changes in the open probability of 4-aminopyridine (4-AP)-sensitive Kv channels defining the outward K⁺ current during phase 1 can modulate Ca²⁺ currents, Ca²⁺ transients, and systolic pressure during the cardiac cycle in intact perfused beating hearts. Pulsed local-field fluorescence microscopy and loose-patch photolysis were used to test the hypothesis that a decrease in a transient K⁺ current (I_to) will enhance Ca²⁺ influx and promote a larger Ca²⁺ transient. Simultaneous recordings of Ca²⁺ transients and APs by pulsed local-field fluorescence microscopy and loose-patch photolysis showed that a reduction in the phase 1 repolarization rate increases the amplitude of Ca²⁺ transients due to an increase in Ca²⁺ influx through L-type Ca²⁺ channels. Moreover, 4-AP induced an increase in the time required for AP to reach 30% repolarization, and the amplitude of Ca²⁺ transients was larger in epicardium than endocardium. On the other hand, the activation of I_to with NS5806 resulted in a reduction of Ca²⁺ current amplitude that led to a reduction of the amplitude of Ca²⁺ transients. Finally, the 4-AP effect on AP phase 1 was significantly smaller when the L-type Ca²⁺ current was partially blocked with nifedipine, indicating that the phase 1 rate of repolarization is defined by the competition between an outward K⁺ current and an inward Ca²⁺ current.