The development of L-type Ca2+ current mediated alternans does not depend on the restitution slope in canine ventricular myocardium

Cardiac alternans have crucial importance in the onset of ventricular fibrillation. The early explanation for alternans development was the voltage-driven mechanism, where the action potential (AP) restitution steepness was considered as crucial determining factor. Recent results suggest that restitution slope is an inadequate predictor for alternans development, but several studies still claim the role of membrane potential as underlying mechanism of alternans. These controversial data indicate that the relationship of restitution and alternans development is not completely understood. APs were measured by conventional microelectrode technique from canine right ventricular papillary muscles. Ionic currents combined with fluorescent measurements were recorded by patch-clamp technique. APs combined with fluorescent measurements were monitored by sharp microelectrodes. Rapid pacing evoked restitution-independent AP duration (APD) alternans. When non-alternating AP voltage command was used, Ca2+i-transient (CaT) alternans were not observed. When alternating rectangular voltage pulses were applied, CaT alternans were proportional to ICaL amplitude alternans. Selective ICaL inhibition did not influence the fast phase of APD restitution. In this study we found that ICaL has minor contribution in shaping the fast phase of restitution curve suggesting that ICaL—if it plays important role in the alternans mechanism—could be an additional factor that attenuates the reliability of APD restitution slope to predict alternans.

Bayesian Calibration of Electrophysiology Models Using Restitution Curve Emulators

Calibration of cardiac electrophysiology models is a fundamental aspect of model personalization for predicting the outcomes of cardiac therapies, simulation testing of device performance for a range of phenotypes, and for fundamental research into cardiac function. Restitution curves provide information on tissue function and can be measured using clinically feasible measurement protocols. We introduce novel “restitution curve emulators” as probabilistic models for performing model exploration, sensitivity analysis, and Bayesian calibration to noisy data. These emulators are built by decomposing restitution curves using principal component analysis and modeling the resulting coordinates with respect to model parameters using Gaussian processes. Restitution curve emulators can be used to study parameter identifiability via sensitivity analysis of restitution curve components and rapid inference of the posterior distribution of model parameters given noisy measurements. Posterior uncertainty about parameters is critical for making predictions from calibrated models, since many parameter settings can be consistent with measured data and yet produce very different model behaviors under conditions not effectively probed by the measurement protocols. Restitution curve emulators are therefore promising probabilistic tools for calibrating electrophysiology models.

Overdrive pacing of spiral waves in a model of human ventricular tissue

High-voltage electrical defibrillation remains the only reliable method of quickly controlling life-threatening cardiac arrhythmias. This paper is devoted to studying an alternative approach, low-voltage cardioversion (LVC), which is based on ideas from non-linear dynamics and aims to remove sources of cardiac arrhythmias by applying high-frequency stimulation to cardiac tissue. We perform a detailed in-silico study of the elimination of arrhythmias caused by rotating spiral waves in a TP06 model of human cardiac tissue. We consider three parameter sets with slopes of the APD restitution curve of 0.7, 1.1 and 1.4, and we study LVC at the baseline and under the blocking of INa and ICaL and under the application of the drugs verapamil and amiodarone. We show that pacing can remove spiral waves; however, its efficiency can be substantially reduced by dynamic instabilities. We classify these instabilities and show that the blocking of INa and the application of amiodarone increase the efficiency of the method, while the blocking of ICaL and the application of verapamil decrease the efficiency. We discuss the mechanisms and the possible clinical applications resulting from our study.

Simultaneous conduction mapping and intracellular membrane potential recording in isolated atria

We describe a novel approach for simultaneously determining regional differences in action potential (AP) morphology and tissue electrophysiological properties in isolated atria. The epicardial surface of rat atrial preparations was placed in contact with a multi-electrode array (9 × 10 silver chloride electrodes, 0.1 mm diameter and 0.1 mm pitch). A glass microelectrode (100 MΩ) was simultaneously inserted into the endocardial surface to record intracellular AP from either of 2 regions (A, B) during pacing from 2 opposite corners of the tissue. AP duration at 80% of repolarisation and its restitution curve was significantly different only in region A (p < 0.01) when AP was initiated at different stimulation sites. Alternans in AP duration and AP amplitude, and in conduction velocity were observed during 2 separate arrhythmic episodes. This approach of combining microelectrode array and intracellular membrane potential recording may provide new insights into arrhythmogenic mechanisms in animal models of cardiovascular disease.

Modelo paramétrico de la actividad eléctrica celular cardiaca estimado a partir de registros electrocardiográficos estándares (Parametrical model of cardiac cell electrical activity estimated from standard electrocardiogram recordings)

El presente trabajo tiene como objetivo principal relacionar la actividad eléctrica cardiaca celular con la actividad eléctrica cardiaca medida en una sola derivación del electrocardiograma (ECG), mediante un modelo paramétrico de potencial de acción (PA) celular, lo cual se llevó a cabo relacionando dinámicas conocidas, matemáticamente modelables, que existen a nivel de una célula cardiaca, a dinámicas que pueden ser encontradas en un registro ECG estándar. La principal dinámica celular a relacionar con el ECG es la conocida como curva de restitución celular en tres dimensiones, la cual relaciona la duración del potencial de acción celular (APD) con el intervalo diastólico que lo precede y con el mismo APD pero del ciclo cardiaco precedente. Curvas de restitución similares se encontraron en señales ECG registradas durante el test isométrico handgrip, relacionando el intervalo QT con el intervalo TQ que lo precede y con el intervalo QT del ciclo cardiaco precedente. Siguiendo esta similitud, un modelo paramétrico de curva de restitución, extraído de un modelo de PA a tres corrientes iónicas, es ajustado a la curva de restitución del ECG con el fin de estimar los parámetros del modelo de PA. Este modelo es finalmente simulado estimulándolo con un tren de impulsos de frecuencia igual a la frecuencia cardiaca del sujeto experimentado. Los resultados muestran que la curva de restitución obtenida experimentalmente a partir del ECG es similar a la obtenida a partir de la simulación del modelo de PA. Más aún, el APD simulado del modelo sigue de forma satisfactoria la variación en el tiempo del intervalo QT del sujeto experimentado. Esto abre nuevas perspectivas en el análisis de la actividad celular a partir de registros ECG estándar.Abstract: The main purpose of this paper is to relate cellular cardiac electrical activity with the cardiac electrical activity measured in only one electrocardiogram (ECG) lead, through a cellular action potential (AP) parametrical model. This is performed by relating known dynamics, which can be mathematically modeled, existing at a cardiac cell level, to dynamics which can be obtained from a standard ECG recording. The main cellular dynamic used for relating with the ECG is the one known as three dimensional cellular restitution curve, which relates the action potential duration (APD) with its preceding diastolic interval and with the APD of the preceding cardiac cycle. Similar restitution curves were found in ECG signals recorded under the isometric handgrip test by relating the QT interval with its preceding TQ interval and with the QT interval of the preceding cardiac cycle. Following this similarity, a parametrical restitution curve, derived from a three ionic current cellular AP model was fitted to the ECG restitution relation for AP model parameter estimation. This model is finally simulated by stimulating it with an impulse train of frequency similar to the heart rate of the tested subject. The results show that the restitution curve experimentally obtained from the ECG is similar to the one obtained from de AP model simulation. Moreover the simulated APD follows satisfactorily the QT interval time variation of the tested subject. This opens new perspectives for the analysis of cellular cardiac electrical activity from standard ECG recordings.

Refractoriness of sarcoplasmic reticulum Ca2+ release determines Ca2+ alternans in atrial myocytes

Cardiac alternans is a recognized risk factor for cardiac arrhythmia and sudden cardiac death. At the cellular level, Ca2+ alternans appears as cytosolic Ca2+ transients of alternating amplitude at regular beating frequency. Cardiac alternans is a multifactorial process but has been linked to disturbances in intracellular Ca2+ regulation. In atrial myocytes, we tested the role of voltage-gated Ca2+ current, sarcoplasmic reticulum (SR) Ca2+ load, and restitution properties of SR Ca2+ release for the occurrence of pacing-induced Ca2+ alternans. Voltage-clamp experiments revealed that peak Ca2+ current was not affected during alternans, and alternans of end-diastolic SR Ca2+ load, evaluated by application of caffeine or measured directly with an intra-SR fluorescent Ca2+ indicator (fluo-5N), were not a requirement for cytosolic Ca2+ alternans. Restitution properties and kinetics of refractoriness of Ca2+ release after activation during alternans were evaluated by four different approaches: measurements of 1) the delay (latency) of occurrence of spontaneous global Ca2+ releases and 2) Ca2+ spark frequency, both during rest after a large and small alternans Ca2+ transient; 3) the magnitude of premature action potential-induced Ca2+ transients after a large and small beat; and 4) the efficacy of a photolytically induced Ca2+ signal (Ca2+ uncaging from DM-nitrophen) to trigger additional Ca2+ release during alternans. The results showed that the latency of global spontaneous Ca2+ release was prolonged and Ca2+ spark frequency was decreased after the large Ca2+ transient during alternans. Furthermore, the restitution curve of the Ca2+ transient elicited by premature action potentials or by photolysis-induced Ca2+ release from the SR lagged behind after a large-amplitude transient during alternans compared with the small-amplitude transient. The data demonstrate that beat-to-beat alternation of the time-dependent restitution properties and refractory kinetics of the SR Ca2+ release mechanism represents a key mechanism underlying cardiac alternans.

The role of dynamic instability and wavelength in arrhythmia maintenance as revealed by panoramic imaging with blebbistatin vs. 2,3-butanedione monoxime

Unlike other excitation-contraction uncouplers, blebbistatin has few electrophysiological side effects and has gained increasing acceptance as an excitation-contraction uncoupler in optical mapping experiments. However, the possible role of blebbistatin in ventricular arrhythmia has hitherto been unknown. Furthermore, experiments with blebbistatin and 2,3-butanedione monoxime (BDM) offer an opportunity to assess the contribution of dynamic instability and wavelength of impulse propagation to the induction and maintenance of ventricular arrhythmia. Recordings of monophasic action potentials were used to assess effects of blebbistatin in Langendorff-perfused rabbit hearts ( n = 5). Additionally, panoramic optical mapping experiments were conducted in rabbit hearts ( n = 7) that were sequentially perfused with BDM, then washed out, and subsequently perfused with blebbistatin. The susceptibility to arrhythmia was investigated using a shock-on-T protocol. We found that 1) application of blebbistatin did not change action potential duration (APD) restitution; 2) in contrast to blebbistatin, BDM flattened APD restitution curve and reduced the wavelength; and 3) incidence of sustained arrhythmia was much lower under blebbistatin than under BDM (2/123 vs. 23/99). While arrhythmias under BDM were able to stabilize, the arrhythmias under blebbistatin were unstable and terminated spontaneously. In conclusion, the lower susceptibility to arrhythmia under blebbistatin than under BDM indicates that blebbistatin has less effects on arrhythmia dynamics. A steep restitution slope under blebbistatin is associated with higher dynamic instability, manifested by the higher incidence of not only wave breaks but also wave extinctions. This relatively high dynamic instability leads to the self-termination of arrhythmia because of the sufficiently long wavelength under blebbistatin.

The effect of cardiac sympathetic denervation through bilateral stellate ganglionectomy on electrical properties of the heart

The role of the cardiac sympathetic nerve activity in various cardiac diseases is typically evaluated using β-adrenergic receptor antagonists. However, these antagonists induce global denervation effects not only in the cardiovascular system, but also in the brain and kidney. The objective of this study was to detect the electrophysiological property changes due to 8 days of cardiac sympathetic denervation and investigate the possible mechanisms underlying these changes using a more cardiac-specific bilateral stellate ganglionectomy (SGX) rat model. High-resolution optical mapping using a voltage-sensitive dye was performed in isolated Langendorff-perfused sham and SGX hearts, which were paced at progressively reduced basic cycle lengths under several different conditions: control, pretreatment with isoproterenol, and administration of atenolol and esmolol. Several electrophysiological parameters were recorded during periodic pacing and ventricular fibrillation (VF). Our results demonstrate that cardiac sympathetic denervation by bilateral SGX shortens action potential duration (APD) and flattens the APD restitution curve, but does not significantly affect spatial dispersion of APD. We found that, although the vulnerability of sham and SGX hearts to VF is similar, the dynamics of VF are different. The maximum dominant frequency is higher, and the spatial distribution of VF is more complex in the SGX heart, resulting in different mechanisms of VF. We demonstrated that β1-adrenergic receptors are downregulated in the SGX compared with sham hearts. In addition, our data suggest that the mechanism of cardiac sympathetic denervation by SGX surgery is more similar to the administration of β-blocker esmolol than atenolol.