Membrane potential determines calcium alternans through modulation of SR Ca²⁺ load and L-type Ca²⁺ current

Alternans is a risk factor for cardiac arrhythmia, including atrial fibrillation. At the cellular level alternans is observed as beat-to-beat alternations in contraction, action potential (AP) morphology and magnitude of the Ca²⁺ transient (CaT). It is widely accepted that the bi-directional interplay between membrane voltage and Ca²⁺ is crucial for the development of alternans, however recently the attention has shifted to instabilities in cellular Ca²⁺ handling, while the role of AP alternation remains poorly understood. This study provides new insights how beat- to-beat alternation in AP morphology affects occurrence of CaT alternans in atrial myocytes. Pacing-induced AP and CaT alternans were studied in rabbit atrial myocytes using combined Ca²⁺ imaging and electrophysiological measurements. To determine the role of AP morphology for the generation of CaT alternans, trains of two voltage commands in form of APs recorded during large and small alternans CaTs were applied to voltage-clamped cells. APs of longer duration (as observed during small amplitude alternans CaT) and especially beat-to-beat alternations in AP morphology (AP alternans) reduced the pacing frequency threshold and increased the degree of CaT alternans. AP morphology contributes to the development of CaT alternans by two mechanisms. First, the AP waveform observed during small alternans CaTs coincided with higher end-diastolic sarcoplasmic reticulum Ca²⁺ levels ([Ca²⁺]_SR), and AP alternans resulted in beat-to-beat alternations in end-diastolic [Ca²⁺]_SR. Second, L-type Ca²⁺ current was significantly affected by AP morphology, where the AP waveform observed during large CaT elicited L-type Ca²⁺ currents of higher magnitude and faster kinetics, resulting in more efficient triggering of SR Ca²⁺ release. In conclusion, alternation in AP morphology plays a significant role in the development and stabilization of atrial alternans. The demonstration that CaT alternans can be controlled or even prevented by modulating AP morphology has important ramifications for arrhythmia prevention and therapy strategies.