Isoproterenol does not enhance Ca-dependent Na/Ca exchange current in intact rabbit ventricular myocytes

Cardiac Na/Ca exchange (NCX, NCX1.1) is critical in cardiac myocyte Ca regulation, and its altered function contributes to inotropic state, systolic dysfunction in heart failure and arrhythmogenesis. Regulation of NCX is multifaceted, but protein kinase A (PKA) effects on NCX function are controversial. Here, we use three different and complementary approaches to compare NCX function ± 1 μM isoproterenol (ISO) in intact rabbit cardiac myocytes (in paired comparisons). First, in field-stimulated intact cells we inferred the cytosolic [Ca] ([Ca]i) dependence of NCX function from the decay rate of caffeine-induced [Ca]i transients. Second, we measured caffeine-induced [Ca]i and inward INCX simultaneously (perforated patch voltage clamp), to measure directly the [Ca]i dependence of NCX rate. Third, using whole cell ruptured patch with [Ca]i heavily buffered to 100 nM, [Na]i = 10 mM, and ICa, SR Ca release and Na/K pump all blocked, we recorded INCX ramps at 37 °C. We find that NCX function is not altered by PKA activation under any of these three protocols, where intracellular conditions ranged from near-physiological to highly controlled. This does not rule out NCX modulation by PKA under all conditions, or in species other than rabbit. However, such effects are likely to be either minor (vs. other PKA actions on myocyte Ca handling) or indirect, such as secondary effects dependent on altered local [Ca]i and [Na]i.