K+ currents in ventricular cardiomyocytes of p.N98S-calmodulin mutant mice.

Missense mutations in calmodulin (CaM)-encoding genes are associated with life-threatening ventricular arrhythmia syndromes. Here, we investigated the role of cardiac K+ channel dysregulation in arrhythmogenic long QT syndrome (LQTS) using a knock-in mouse model heterozygous for a recurrent mutation (p.N98S) in the Calm1 gene (Calm1N98S/+). Single-cell patch-clamp technique and whole heart optical voltage mapping were used to assess action potentials and whole cell currents. Ventricular action potential duration (APD) at baseline was similar between genotypes. The β-adrenergic agonist isoproterenol prolonged APD in myocytes and isolated perfused hearts from Calm1N98S/+, but not wild-type (Calm1+/+), mice. Current density-voltage relationships for the small-conductance calcium-activated K+ (SK) current and the inward rectifier K+ current did not significantly differ between Calm1+/+ and Calm1N98S/+ ventricular cardiomyocytes ± isoproterenol. Peak densities of other voltage-gated K+ currents were significantly larger in Calm1N98S/+ versus Calm1+/+ cells at voltages ≥40 mV, both without and with isoproterenol. Isoproterenol reduced outward KATP currents more in Calm1N98S/+ versus Calm1+/+ myocytes. Dialysis of Calm1+/+ cardiomyocytes with exogenous wild-type or N98S-CaM protein (5 µmol/L) via the pipette, respectively, increased and eliminated SK currents. The specific SK channel inhibitor apamin did not significantly alter the APD of Calm1+/+ or Calm1N98S/+ hearts ± isoproterenol. Thus, dysregulation of SK or voltage-gated K+ channels does not contribute to the β-adrenergic-induced LQTS of Calm1N98S/+ mice, possibly because cardiomyocyte content of endogenous N98S-CaM and/or its affinity for CaM-binding domains may be too low to modulate channel properties. The larger KATP current inhibition by isoproterenol may delay Calm1N98S/+ myocyte repolarization at low intracellular [ATP].NEW & NOTEWORTHY Despite in vitro and in silico evidence implicating cardiac K+ channel dysregulation in LQTS associated with missense mutations in genes-encoding calmodulin, their effects on native cardiac K+ currents are unknown. Using a knock-in mouse model harboring the p.N98S mutation in the Calm1 gene, we found no evidence for dysregulation of major cardiac K+ channels. Although these data do not support mechanistic findings from heterologous systems, our finding impacts efforts to improve therapies for calmodulinopathies.