Quantitative analysis of the Ca²⁺ -dependent regulation of delayed rectifier K⁺ current I_Ks in rabbit ventricular myocytes

Key points: [Ca²⁺ ]_i enhanced rabbit ventricular slowly activating delayed rectifier K⁺ current (I_Ks ) by negatively shifting the voltage dependence of activation and slowing deactivation, similar to perfusion of isoproterenol. Rabbit ventricular rapidly activating delayed rectifier K⁺ current (I_Kr ) amplitude and voltage dependence were unaffected by high [Ca²⁺ ]_i . When measuring or simulating I_Ks during an action potential, I_Ks was not different during a physiological Ca²⁺ transient or when [Ca²⁺ ]_i was buffered to 500 nm.

Abstract: The slowly activating delayed rectifier K⁺ current (I_Ks ) contributes to repolarization of the cardiac action potential (AP). Intracellular Ca²⁺ ([Ca²⁺ ]_i ) and β-adrenergic receptor (β-AR) stimulation modulate I_Ks amplitude and kinetics, but details of these important I_Ks regulators and their interaction are limited. We assessed the [Ca²⁺ ]_i dependence of I_Ks in steady-state conditions and with dynamically changing membrane potential and [Ca²⁺ ]_i during an AP. I_Ks was recorded from freshly isolated rabbit ventricular myocytes using whole-cell patch clamp. With intracellular pipette solutions that controlled free [Ca²⁺ ]_i , we found that raising [Ca²⁺ ]_i from 100 to 600 nm produced similar increases in I_Ks as did β-AR activation, and the effects appeared additive. Both β-AR activation and high [Ca²⁺ ]_i increased maximally activated tail I_Ks , negatively shifted the voltage dependence of activation, and slowed deactivation kinetics. These data informed changes in our well-established mathematical model of the rabbit myocyte. In both AP-clamp experiments and simulations, I_Ks recorded during a normal physiological Ca²⁺ transient was similar to I_Ks measured with [Ca²⁺ ]_i clamped at 500-600 nm. Thus, our study provides novel quantitative data as to how physiological [Ca²⁺ ]_i regulates I_Ks amplitude and kinetics during the normal rabbit AP. Our results suggest that micromolar [Ca²⁺ ]_i , in the submembrane or junctional cleft space, is not required to maximize [Ca²⁺ ]_i -dependent I_Ks activation during normal Ca²⁺ transients.