Impact of ISK Voltage and Ca2+/Mg2+-Dependent Rectification on Cardiac Repolarization

Abstract

Cardiac small conductance Ca²⁺-activated K⁺ (SK) channels are activated solely by Ca²⁺, but the SK current (I_SK) is inwardly rectified. However, the impact of inward rectification in shaping action potentials (APs) in ventricular cardiomyocytes under β-adrenergic stimulation or in disease states remains undefined. Two processes underlie this inward rectification: an intrinsic rectification caused by an electrostatic energy barrier from positively charged amino acids at the inner pore and a voltage-dependent Ca²⁺/Mg²⁺ block. Thus, Ca²⁺ has a biphasic effect on I_SK, activating at low [Ca²⁺] yet inhibiting I_SK at high [Ca²⁺]. We examined the effect of I_SK rectification on APs in rat cardiomyocytes by simultaneously recording whole-cell apamin-sensitive currents and Ca²⁺ transients during an AP waveform and developed a computer model of SK channels with rectification features. The typical profile of I_SK during AP clamp included an initial peak (mean 1.6 pA/pF) followed by decay to the point that submembrane [Ca²⁺] reached ∼10 μM. During the rest of the AP stimulus, I_SK either plateaued or gradually increased as the cell repolarized and submembrane [Ca²⁺] decreased further. We used a six-state gating model combined with intrinsic and Ca²⁺/Mg²⁺-dependent rectification to simulate I_SK and investigated the relative contributions of each type of rectification to AP shape. This SK channel model replicates key features of I_SK recording during AP clamp showing that intrinsic rectification limits I_SK at high V_m during the early and plateau phase of APs. Furthermore, the initial rise of Ca²⁺ transients activates, but higher [Ca²⁺] blocks SK channels, yielding a transient outward-like I_SK trajectory. During the decay phase of Ca²⁺, the Ca²⁺-dependent block is released, causing I_SK to rise again and contribute to repolarization. Therefore, I_SK is an important repolarizing current, and the rectification characteristics of an SK channel determine its impact on early, plateau, and repolarization phases of APs.