Cellular mechanisms of ventricular arrhythmias in a mouse model of Timothy syndrome (long QT syndrome 8)

Ca2+ flux through L-type Ca(V)1.2 channels shapes the waveform of the ventricular action potential (AP) and is essential for excitation-contraction (EC) coupling. Timothy syndrome (TS) is a disease caused by a gain-of-function mutation in the Ca(V)1.2 channel (Ca(V)1.2-TS) that decreases inactivation of the channel, which increases Ca2+ influx, prolongs APs, and causes lethal arrhythmias. Although many details of the Ca(V)1.2-TS channels are known, the cellular mechanisms by which they induce arrhythmogenic changes in intracellular Ca2+ remain unclear. We found that expression of Ca(V)1.2-TS channels increased sarcolemmal Ca2+ "leak" in resting TS ventricular myocytes. This resulted in higher diastolic [Ca2+]; in TS ventricular myocytes compared to WT. Accordingly, TS myocytes had higher sarcoplasmic reticulum (SR) Ca2+ load and Ca2+ spark activity, larger amplitude [Ca2+]; transients, and augmented frequency of Ca2 waves. The large SR Ca2 release in TS myocytes had a profound effect on the kinetics of Ca(V)1.2 current in these cells, increasing the rate of inactivation to a high, persistent level. This limited the amount of influx during EC coupling in TS myocytes. The relationship between the level of expression of Ca(V)1.2-TS channels and the probability of Ca2 wave occurrence was non-linear, suggesting that even low levels of these channels were sufficient to induce maximal changes in [Ca2+](i). Depolarization of WT cardiomyocytes with a TS AP waveform increased, but did not equalize [Ca2+](i), compared to depolarization of TS myocytes with the same waveform. We propose that Ca(V)1.2-TS channels increase [Ca2+] in the cytosol and the SR, creating a Ca2+ overloaded state that increases the probability of arrhythmogenic spontaneous SR Ca2+ release. (C) 2013 Elsevier Ltd. All rights reserved.