UC Davis

Late Na⁺ current (I_NaL) significantly contributes to shaping cardiac action potentials (APs) and increased I_NaL is associated with cardiac arrhythmias. β-adrenergic receptor (βAR) stimulation and its downstream signaling via protein kinase A (PKA) and Ca²⁺/calmodulin-dependent protein kinase II (CaMKII) pathways are known to regulate I_NaL. However, it remains unclear how each of these pathways regulates I_NaL during the AP under physiological conditions. Here we performed AP-clamp experiments in rabbit ventricular myocytes to delineate the impact of each signaling pathway on I_NaL at different AP phases to understand the arrhythmogenic potential. During the physiological AP (2 Hz, 37 °C) we found that I_NaL had a basal level current independent of PKA, but partially dependent on CaMKII. βAR activation (10 nM isoproterenol, ISO) further enhanced I_NaL via both PKA and CaMKII pathways. However, PKA predominantly increased I_NaL early during the AP plateau, whereas CaMKII mainly increased I_NaL later in the plateau and during rapid repolarization. We also tested the role of key signaling pathways through exchange protein activated by cAMP (Epac), nitric oxide synthase (NOS) and reactive oxygen species (ROS). Direct Epac stimulation enhanced I_NaL similar to the βAR-induced CaMKII effect, while NOS inhibition prevented the βAR-induced CaMKII-dependent I_NaL enhancement. ROS generated by NADPH oxidase 2 (NOX2) also contributed to the ISO-induced I_NaL activation early in the AP. Taken together, our data reveal differential modulations of I_NaL by PKA and CaMKII signaling pathways at different AP phases. This nuanced and comprehensive view on the changes in I_NaL during AP deepens our understanding of the important role of I_NaL in reshaping the cardiac AP and arrhythmogenic potential under elevated sympathetic stimulation, which is relevant for designing therapeutic treatment of arrhythmias under pathological conditions.