ERG potassium channels and T‐type calcium channels contribute to the pacemaker and atrioventricular conduction in zebrafish larvae

Abstract

AimBradyarrhythmias result from inhibition of automaticity, prolonged repolarization, or slow conduction in the heart. The ERG channels mediate the repolarizing current IKr in the cardiac action potential, whereas T‐type calcium channels (TTCC) are involved in the sinoatrial pacemaker and atrioventricular conduction in mammals. Zebrafish have become a valuable research model for human cardiac electrophysiology and disease. Here, we investigate the contribution of ERG channels and TTCCs to the pacemaker and atrioventricular conduction in zebrafish larvae and determine the mechanisms causing atrioventricular block.MethodsZebrafish larvae expressing ratiometric fluorescent Ca2+ biosensors in the heart were used to measure Ca2+ levels and rhythm in beating hearts in vivo, concurrently with contraction and hemodynamics. The atrioventricular delay (the time between the start of atrial and ventricular Ca2+ transients) was used to measure impulse conduction velocity and distinguished between slow conduction and prolonged refractoriness as the cause of the conduction block.ResultsERG blockers caused bradycardia and atrioventricular block by prolonging the refractory period in the atrioventricular canal and in working ventricular myocytes. In contrast, inhibition of TTCCs caused bradycardia and second‐degree block (Mobitz type I) by slowing atrioventricular conduction. TTCC block did not affect ventricular contractility, despite being highly expressed in cardiomyocytes. Concomitant measurement of Ca2+ levels and ventricular size showed mechano‐mechanical coupling: increased preload resulted in a stronger heart contraction in vivo.ConclusionERG channels and TTCCs influence the heart rate and atrioventricular conduction in zebrafish larvae. The zebrafish lines expressing Ca2+ biosensors in the heart allow us to investigate physiological feedback mechanisms and complex arrhythmias.

Suppression of Contraction Raises Calcium Ion Levels in the Heart of Zebrafish Larvae

“…These morphants also presented defects in trabeculation and valve formation [31,32]. Since trabeculae act as an early conduction system in the larval ventricle, the atrioventricular conduction blocks observed in some tnnt2a morphant hearts at 3 and 5 dpf (Figure 1F,G) could be due to a malfunction of ventricular electrical excitation or a block in the atrioventricular canal due to altered action potentials [33].…”

“…Thus, cardiac contraction appears to be an important regulator of systolic and diastolic Ca 2+ levels. In contrast to electromechanically uncoupled hearts, optical mapping studies in unconstrained contracting hearts allow the study of interactions between contraction, electrophysiology, and metabolism, and enhance the translational applicability of results [10,19,33,34].…”