Enhanced cardiac late Na current (late I_Na) and increased sarcoplasmic reticulum (SR)-Ca²⁺-leak are both highly arrhythmogenic. This study seeks to identify signalling pathways interconnecting late I_Na and SR-Ca²⁺-leak in atrial cardiomyocytes (CMs).

In murine atrial CMs, SR-Ca²⁺-leak was increased by the late I_Na enhancer Anemonia sulcata toxin II (ATX-II). An inhibition of Ca²⁺/calmodulin-dependent protein kinase II (Autocamide-2-related inhibitory peptide), protein kinase A (H89), or late I_Na (Ranolazine or Tetrodotoxin) all prevented ATX-II-dependent SR-Ca²⁺-leak. The SR-Ca²⁺-leak induction by ATX-II was not detected when either the Na⁺/Ca²⁺ exchanger was inhibited (KBR) or in CaMKIIδc-knockout mice. FRET measurements revealed increased cAMP levels upon ATX-II stimulation, which could be prevented by inhibition of adenylyl cyclases (ACs) 5 and 6 (NKY 80) but not by inhibition of phosphodiesterases (IBMX), suggesting PKA activation via an AC-dependent increase of cAMP levels. Western blots showed late I_Na-dependent hyperphosphorylation of CaMKII as well as PKA target sites at ryanodine receptor type-2 (-S2814 and -S2808) and phospholamban (-Thr17, -S16). Enhancement of late I_Na did not alter Ca²⁺-transient amplitude or SR-Ca²⁺-load. However, upon late I_Na activation and simultaneous CaMKII inhibition, Ca²⁺-transient amplitude and SR-Ca²⁺-load were increased, whereas PKA inhibition reduced Ca²⁺-transient amplitude and load and additionally slowed Ca²⁺ elimination. In atrial CMs from patients with atrial fibrillation, inhibition of late I_Na, CaMKII, or PKA reduced the SR-Ca²⁺-leak.

Late I_Na exerts distinct effects on Ca²⁺ homeostasis in atrial myocardium through activation of CaMKII and PKA. Inhibition of late I_Na represents a potential approach to attenuate CaMKII activation and decreases SR-Ca²⁺-leak in atrial rhythm disorders. The interconnection with the cAMP/PKA system further increases the antiarrhythmic potential of late I_Na inhibition.