Calcium-Handling Abnormalities Underlying Atrial Arrhythmogenesis and Contractile Dysfunction in Dogs With Congestive Heart Failure

Abstract

Background— Congestive heart failure (CHF) is a common cause of atrial fibrillation. Focal sources of unknown mechanism have been described in CHF-related atrial fibrillation. The authors hypothesized that abnormal calcium (Ca 2+ ) handling contributes to the CHF-related atrial arrhythmogenic substrate. Methods and Results— CHF was induced in dogs by ventricular tachypacing (240 bpm �2 weeks). Cellular Ca 2+ -handling properties and expression/phosphorylation status of key Ca 2+ handling and myofilament proteins were assessed in control and CHF atria. CHF decreased cell shortening but increased left atrial diastolic intracellular Ca 2+ concentration ([Ca 2+ ] i ), [Ca 2+ ] i transient amplitude, and sarcoplasmic reticulum (SR) Ca 2+ load (caffeine-induced [Ca 2+ ] i release). SR Ca 2+ overload was associated with spontaneous Ca 2+ transient events and triggered ectopic activity, which was suppressed by the inhibition of SR Ca 2+ release (ryanodine) or Na + /Ca 2+ exchange. Mechanisms underlying abnormal SR Ca 2+ handling were then studied. CHF increased atrial action potential duration and action potential voltage clamp showed that CHF-like action potentials enhance Ca 2+ i loading. CHF increased calmodulin-dependent protein kinase II phosphorylation of phospholamban by 120%, potentially enhancing SR Ca 2+ uptake by reducing phospholamban inhibition of SR Ca 2+ ATPase, but it did not affect phosphorylation of SR Ca 2+ -release channels (RyR2). Total RyR2 and calsequestrin (main SR Ca 2+ -binding protein) expression were significantly reduced, by 65% and 15%, potentially contributing to SR dysfunction. CHF decreased expression of total and protein kinase A–phosphorylated myosin-binding protein C (a key contractile filament regulator) by 27% and 74%, potentially accounting for decreased contractility despite increased Ca 2+ transients. Complex phosphorylation changes were explained by enhanced calmodulin-dependent protein kinase IIδ expression and function and type-1 protein-phosphatase activity but downregulated regulatory protein kinase A subunits. Conclusions— CHF causes profound changes in Ca 2+ -handling and -regulatory proteins that produce atrial fibrillation–promoting atrial cardiomyocyte Ca 2+ -handling abnormalities, arrhythmogenic triggered activity, and contractile dysfunction.