Ion current profiles in canine ventricular myocytes obtained by the "onion peeling" technique

The profiles of ion currents during the cardiac action potential can be visualized by the action potential voltage clamp technique. To obtain multiple ion current data from the same cell, the "onion peeling" technique, based on sequential pharmacological dissection of ion currents, has to be applied. Combination of the two methods allows recording of several ion current profiles from the same myocyte under largely physiological conditions. Using this approach, we have studied the densities and integrals of the major cardiac inward (I_Ca, I_NCX, I_Na-late) and outward (I_Kr, I_Ks, I_K1) currents in canine ventricular cells and studied the correlation between them. For this purpose, canine ventricular cardiomyocytes were chosen because their electrophysiological properties are similar to those of human ones. Significant positive correlation was observed between the density and integral of I_Ca and I_Kr, and positive correlation was found also between the integral of I_Ca and I_NCX. No further correlations were detected. The Ca²⁺-sensitivity of K⁺ currents was studied by comparing their parameters in the case of normal calcium homeostasis and following blockade of I_Ca. Out of the three K⁺ currents studied, only I_Ks was Ca²⁺-sensitive. The density and integral of I_Ks was significantly greater, while its time-to-peak value was shorter at normal Ca²⁺ cycling than following I_Ca blockade. No differences were detected for I_Kr or I_K1 in this regard. Present results indicate that the positive correlation between I_Ca and I_Kr prominently contribute to the balance between inward and outward fluxes during the action potential plateau in canine myocytes. The results also suggest that the profiles of cardiac ion currents have to be studied under physiological conditions, since their behavior may strongly be influenced by the intracellular Ca²⁺ homeostasis and the applied membrane potential protocol.