Activation of reverse Na+-Ca2 + exchanger by skeletal Na+ channel isoform increases excitation-contraction coupling efficiency in rabbit cardiomyocytes

Our prior work has shown that Na⁺ current (I_Na) affects sarcoplasmic reticular (SR) CA²⁺ release by activating early reverse of the Na⁺-Ca²⁺ exchanger (NCX). The resulting CA²⁺ entry primes the dyadic cleft, which appears to increase CA²⁺ channel coupling fidelity. It has been shown that the skeletal isoform of the voltage-gated Na⁺ channel (Na_v1.4) is the main tetrodotoxin (TTX)-sensitive Na_v isoform expressed in adult rabbit ventricular cardiomyocytes. Here, I tested the hypothesis that it is also the principal isoform involved in the priming mechanism. Action potentials (APs) were evoked in isolated rabbit ventricular cells loaded with fluo-4, and simultaneously recorded CA²⁺ transients before and after the application of either relatively low doses of TTX (100 nM), the specific Na_v1.4 inhibitor μ-Conotoxin GIIIB or the specific Na_v1.1 inhibitor ICA 121430. Although APs changes after the application of each drug reflected the relative abundance of each isoform, the effects of TTX and GIIIB on SR CA²⁺ release (measured as the transient maximum upstroke velocity) were no different. Furthermore, this reduction in SR CA²⁺ release was comparable with the value that we obtained previously when total I_Na was inactivated with a ramp applied under voltage clamp. Finally, SR CA²⁺ release was unaltered by the same ramp in the presence of TTX or GIIB. In contrast, application of ICA had no effect of SR CA²⁺ release. These results suggest that Na_v1.4 is the main Na_v isoform involved in regulating the efficiency of excitation-contraction coupling in rabbit cardiomyocytes by priming the junction via activation of reverse-mode NCX.NEW & NOTEWORTHY A number of studies suggest that the Na⁺-Ca²⁺ exchanger (NCX) activated by Na⁺ currents is involved in the process of excitation-contraction (EC) coupling in cardiac ventricular myocytes. Although insufficient to trigger sarcoplasmic CA²⁺ release alone, the CA²⁺ entering through reverse NCX during an action potential can prime the dyadic cleft and increase the CA²⁺ current coupling fidelity. Using specific Na⁺ inhibitors in this study, we show that in rabbit ventricular cells the skeletal Na⁺ channel isoform (Na_v1.4) is the main isoform responsible for this priming. Our study provides insights into a mechanism that may have an increased relevance where EC coupling is remodeled.

Ca2+-induced Ca2+ release; Na+ channels; Na+-Ca2+ exchanger; cardiac excitation-contraction coupling.