Ionic mechanisms of electrophysiological properties and repolarization abnormalities in rabbit Purkinje fibers

摘要

Purkinje cells play an important role in drag-induced anhythmogenesis and are widely used in preclinical drag safety assessments. Repolarization abnormalities such as action potential (AP) prolongation and early afterdeploarizations (EAD) are often observed in vitro upon pharmacological interventions. However, because drugs do not act on only one defined target, it is often difficult to fully explain the mechanisms of action and their potential arrhythmogenicity. Computational models, when appropriately detailed and validated, can be used to gain mechanistic insights into the mechanisms of action of certain drags. Nevertheless, no model of Purkinje electrophysiology that is able to reproduce characteristic Purkinje responses to drag-induced changes in ionic current conductances such as AP prolongation and EAD generation currently exists. In this study, a novel biophysically detailed model of rabbit Purkinje electrophysiology was developed by integration of data from voltage-clamp and AP experimental recordings. Upon validation, we demonstrate that the model reproduces many key electrophysiological properties of rabbit Purkinje cells. These include: AP morphology and duration, both input resistance and rate dependence properties as well as response to hyperkalemia. Pharmacological interventions such as inward rectifier K~+ current and rapid delayed rectifier K~+ current block as well as late Na~+ current increase result in significant AP changes. However, enhanced L-type Ca~(2+) current i_(caL) dominates in EAD genesis in Purkinje fibers. In addition, i_(caL) inactivation dynamics and intercellular coupling in tissue strongly modulate EAD formation. We conclude that EAD generation in Purkinje cells is mediated by an increase in i_(caL) and modulated by its inactivation kinetics.