Ablation of sites displaying Purkinje activity is highly effective against idiopathic ventricular fibrillation which often originates in the right ventricular outflow tract. However, during endocardial mapping Purkinje potentials are rarely, if never, detected in the right ventricular outflow tract. In the present study, we aimed to determine whether the Purkinje system extends into the right ventricular outflow tract. Hearts of five female sheep were blood-perfused in a Langendorff setup in which we performed epicardial and endocardial voltage mapping. During atrial pacing, the right ventricular outflow tract epicardium activated later than the epicardium of the left and right ventricular free walls. Endocardial mapping revealed Purkinje spikes at several sites in the free wall of the right ventricular outflow tract. In one heart, Purkinje spikes preceded ventricular premature beats during mapping, but were not visible during sinus rhythm. Subsequent immuno-histological examination showed a network of Connexin 40-positive Purkinje fibers across and within the wall of the right ventricular outflow tract. Quantitative analysis revealed that the transmural Purkinje fiber network was more abundant near the endocardium than epicardium. In conclusion, the Purkinje system extends into the right ventricular outflow tract of the sheep heart. These findings demonstrate that the sheep could be a valuable model for studying Purkinje-related arrhythmias in the right ventricular outflow tract.

To unlock new research possibilities by acquiring control of action potential (AP) morphologies in excitable cells, we developed an opto-electronic feedback loop-based system integrating cellular electrophysiology, real-time computing, and optogenetic approaches and applied it to monolayers of heart muscle cells. This allowed accurate restoration and preservation of cardiac AP morphologies in the presence of electrical perturbations of different origin in an unsupervised, self-regulatory manner, without any prior knowledge of the disturbance. Moreover, arbitrary AP waveforms could be enforced onto these cells. Collectively, these results set the stage for the refinement and application of opto-electronic control systems to enable in-depth investigation into the regulatory role of membrane potential in health and disease.