Voltage Clocks Research Articles

RyR2R420Q catecholaminergic polymorphic ventricular tachycardia mutation induces bradycardia by disturbing the coupled clock pacemaker mechanism

Catecholaminergic polymorphic ventricular tachycardia (CPVT) is a lethal genetic arrhythmia that manifests syncope or sudden death in children and young adults under stress conditions. CPVT patients often present bradycardia and sino-atrial node (SAN) dysfunction. However, the mechanism remains unclear. We analyzed SAN function in two CPVT families and in a novel knock-in (KI) mouse model carrying the RyR2R420Q mutation. Humans and KI mice presented slower resting heart rate. Accordingly, the rate of spontaneous intracellular Ca2+ ([Ca2+]i) transients was slower in KI mouse SAN preparations than in WT, without any significant alteration in the "funny" current (If ). The L-type Ca2+ current was reduced in KI SAN cells in a [Ca2+]i-dependent way, suggesting that bradycardia was due to disrupted crosstalk between the "voltage" and "Ca2+" clock, and the mechanisms of pacemaking was induced by aberrant spontaneous RyR2- dependent Ca2+ release. This finding was consistent with a higher Ca2+ leak during diastolic periods produced by long-lasting Ca2+ sparks in KI SAN cells. Our results uncover a mechanism for the CPVT-causing RyR2 N-terminal mutation R420Q, and they highlight the fact that enhancing the Ca2+ clock may slow the heart rhythm by disturbing the coupling between Ca2+ and voltage clocks.

Improved Model of the Coupled Ca2+ and Sarcolemmal Voltage Clocks for Sinoatrial Node Cell

Improved Model of the Coupled Ca2+ and Sarcolemmal Voltage Clocks for Sinoatrial Node Cell

Mathematical Modelling of the Autonomous Activity of Cultured Neonatal Rat Ventricular Myocytes

Problematic. The biological pacemaker is a new therapeutic approach that could lead to optimized treatment of bradycardia. A possibility is the development of a thin sheet of cardiomyocytes, cultured to obtain a target activation rate. Fundamental research, often conducted with neonatal rat ventricular myocytes (NRVMs), partially revealed two basic coupled mechanisms of automaticity termed Voltage Clock (synergy of membrane currents) and Calcium Clock (internal oscillations of calcium concentration). To date, no ionic model is able to reproduce in silico the autonomous activity found in cultured NRVMs. The present project aims to fill this gap.Methods. A non-automatic NRVM ionic model (Korhonen-Tavi, 2009) is modified according to documented Voltage and Calcium Clocks mathematical formulations. The myocytes are cultured for 48 hours at low density, allowing cells to remain single on dishes. Autonomous action potentials (APs) are measured with patch clamp method, and calcium transients (CTs) with Fluo-4 AM fluorescence imaging. Experimental variables (spontaneous rate, AP amplitude and duration, CT decay time course) are extracted and compared to the output of the model. Results. Insertion of Voltage Clock (upregulation of If and ICaL, and downregulation of IK1 and Ito) into the model showed the presence of automaticity with the characteristic slow late diastole depolarization. The maximum activation frequencies (0,7Hz) reached only the lower frequency range documented experimentally and found in the litterature (0,4 to 6Hz). Conclusion. Limitations in the rate of automaticity found emphasise the importance of exploring the Calcium Clock mechanism whose insertion may lead to higher spontaneous rates. In the future, a bifurcation analysis will give some insights on the dynamical mechanisms behind pacemaker generation in cultured NRVMs.

Low Energy Magnetic Domain Wall Logic in Short, Narrow, Ferromagnetic Wires

We present circuit simulation results of an implementation of universal logic that operates at low switching energy. Information is stored in the position of a single domain wall in a thin, short ferromagnetic wire. The gate is switched by current-driven domain wall motion, and information is read out using a magnetic tunnel junction. The inputs and outputs of the device are currents controlled by voltage clocks, making it compatible with CMOS. Using devices that operate at 100-1 mV, we simulate a shift register circuit and a full-adder circuit. The simulations show that the magnetic logic gates can operate at lower switching energy than CMOS electronics.

Robust Level Converter for Sub-Threshold/Super-Threshold Operation:100 mV to 2.5 V

For ultra low power application, digital sub-threshold logic design has been explored. Extremely low power supply (VDD) of sub-threshold logic results in significant power reduction. However, it is difficult to convert signals from core logic to input/output (I/O) circuits since core VDD is vastly different from high I/O supply voltage. In this work, we propose a level converter based on dynamic logic style for sub-threshold I/O part, having a large dynamic range of conversion. For the level converter, high voltage clock signal needs to be delivered through separate clock path from core logic, leading to clock synchronization problem between high voltage and low voltage clocks. To overcome this issue, we employed a Clock Synchronizer. A test chip is fabricated in 130-nm CMOS technology in order to verify the proposed technique. Hardware measurement results show that the level converter successfully converts 0.3 V 8 MHz pulse to 2.5 V signal.

The Role of the Calcium and the Voltage Clocks in Sinoatrial Node Dysfunction

Recent evidence indicates that the voltage clock (cyclic activation and deactivation of membrane ion channels) and Ca2+ clocks (rhythmic spontaneous sarcoplasmic reticulum Ca2+ release) jointly regulate sinoatrial node (SAN) automaticity. However, the relative importance of the voltage clock and Ca2+ clock for pacemaking was not revealed in sick sinus syndrome. Previously, we mapped the intracellular calcium (Cai) and membrane potentials of the normal intact SAN simultaneously using optical mapping in Langendorff-perfused canine right atrium. We demonstrated that the sinus rate increased and the leading pacemaker shifted to the superior SAN with robust late diastolic Cai elevation (LDCAE) during β-adrenergic stimulation. We also showed that the LDCAE was caused by spontaneous diastolic sarcoplasmic reticulum (SR) Ca2+ release and was closely related to heart rate changes. In contrast, in pacing induced canine atrial fibrillation and SAN dysfunction models, Ca2+ clock of SAN was unresponsiveness to β-adrenergic stimulation and caffeine. Ryanodine receptor 2 (RyR2) in SAN was down-regulated. Using the prolonged low dose isoproterenol together with funny current block, we produced a tachybradycardia model. In this model, chronically elevated sympathetic tone results in abnormal pacemaking hierarchy in the right atrium, including suppression of the superior SAN and enhanced pacemaking from ectopic sites. Finally, if the LDCAE was too small to trigger an action potential, then it induced only delayed afterdepolarization (DAD)-like diastolic depolarization (DD). The failure of DAD-like DD to consistently trigger a sinus beat is a novel mechanism of atrial arrhythmogenesis. We conclude that dysfunction of both the Ca2+ clock and the voltage clock are important in sick sinus syndrome.

A coupled SYSTEM of intracellular Ca2+ clocks and surface membrane voltage clocks controls the timekeeping mechanism of the heart's pacemaker.

Ion channels on the surface membrane of sinoatrial nodal pacemaker cells (SANCs) are the proximal cause of an action potential. Each individual channel type has been thoroughly characterized under voltage clamp, and the ensemble of the ion channel currents reconstructed in silico generates rhythmic action potentials. Thus, this ensemble can be envisioned as a surface "membrane clock" (M clock). Localized subsarcolemmal Ca(2+) releases are generated by the sarcoplasmic reticulum via ryanodine receptors during late diastolic depolarization and are referred to as an intracellular "Ca(2+) clock," because their spontaneous occurrence is periodic during voltage clamp or in detergent-permeabilized SANCs, and in silico as well. In spontaneously firing SANCs, the M and Ca(2+) clocks do not operate in isolation but work together via numerous interactions modulated by membrane voltage, subsarcolemmal Ca(2+), and protein kinase A and CaMKII-dependent protein phosphorylation. Through these interactions, the 2 subsystem clocks become mutually entrained to form a robust, stable, coupled-clock system that drives normal cardiac pacemaker cell automaticity. G protein-coupled receptors signaling creates pacemaker flexibility, ie, effects changes in the rhythmic action potential firing rate, by impacting on these very same factors that regulate robust basal coupled-clock system function. This review examines evidence that forms the basis of this coupled-clock system concept in cardiac SANCs.

Beating to time: calcium clocks, voltage clocks, and cardiac pacemaker activity

the heartbeat arises in the sinoatrial node (SAN) and then spreads throughout the heart. SAN function is therefore essential for normal cardiac physiology. The function of the SAN decreases with age, and this has been correlated with changes of ion channel expression ([12][1]). It is therefore of

Synergism of coupled subsarcolemmal Ca2+clocks and sarcolemmal voltage clocks confers robust and flexible pacemaker function in a novel pacemaker cell model

Recent experimental studies have demonstrated that sinoatrial node cells (SANC) generate spontaneous, rhythmic, local subsarcolemmal Ca(2+) releases (Ca(2+) clock), which occur during late diastolic depolarization (DD) and interact with the classic sarcolemmal voltage oscillator (membrane clock) by activating Na(+)-Ca(2+) exchanger current (I(NCX)). This and other interactions between clocks, however, are not captured by existing essentially membrane-delimited cardiac pacemaker cell numerical models. Using wide-scale parametric analysis of classic formulations of membrane clock and Ca(2+) cycling, we have constructed and initially explored a prototype rabbit SANC model featuring both clocks. Our coupled oscillator system exhibits greater robustness and flexibility than membrane clock operating alone. Rhythmic spontaneous Ca(2+) releases of sarcoplasmic reticulum (SR)-based Ca(2+) clock ignite rhythmic action potentials via late DD I(NCX) over much broader ranges of membrane clock parameters [e.g., L-type Ca(2+) current (I(CaL)) and/or hyperpolarization-activated ("funny") current (I(f)) conductances]. The system Ca(2+) clock includes SR and sarcolemmal Ca(2+) fluxes, which optimize cell Ca(2+) balance to increase amplitudes of both SR Ca(2+) release and late DD I(NCX) as SR Ca(2+) pumping rate increases, resulting in a broad pacemaker rate modulation (1.8-4.6 Hz). In contrast, the rate modulation range via membrane clock parameters is substantially smaller when Ca(2+) clock is unchanged or lacking. When Ca(2+) clock is disabled, the system parametric space for fail-safe SANC operation considerably shrinks: without rhythmic late DD I(NCX) ignition signals membrane clock substantially slows, becomes dysrhythmic, or halts. In conclusion, the Ca(2+) clock is a new critical dimension in SANC function. A synergism of the coupled function of Ca(2+) and membrane clocks confers fail-safe SANC operation at greatly varying rates.

The calcium and voltage clocks in sinoatrial node automaticity.

Recent evidence indicates that the voltage (cyclic activation and deactivation of membrane ion channels) and Ca2+ clocks (rhythmic spontaneous sarcoplasmic reticulum Ca2+ release) jointly regulate sinoatrial node (SAN) automaticity. Since the intact SAN is a heterogeneous structure that includes multiple different cell types interacting with each other, the relative importance of the voltage and Ca2+ clocks for pacemaking may be variable in different regions of the SAN. Recently, we performed optical mapping in isolated and Langendorff-perfused canine right atria. We mapped the intracellular calcium (Cai) and membrane potentials of the intact SAN simultaneously. Using previously described criteria of the timing of the late diastolic Cai elevation (LDCAE) relative to the action potential upstroke to detect Ca2+ clock activity, we demonstrated that the sinus rate increased and the leading pacemaker shifted to the superior SAN with the robust LDCAE during β-adrenergic stimulation. We also showed that the LDCAE was caused by spontaneous diastolic SR Ca2+ release and was closely related with heart rate changes. We conclude that the Ca2+ and voltage clocks work synergistically to generate SAN automaticity.

TONUS CHANGES IN CARDIAC MUSCLE AND THEIR SIGNIFICANCE FOR THE INITIATION OF IMPULSES

ArticleTONUS CHANGES IN CARDIAC MUSCLE AND THEIR SIGNIFICANCE FOR THE INITIATION OF IMPULSESEmil BozlerEmil BozlerFrom the Department of Physiology, Ohio State UniversityPublished Online:01 Jul 1943https://doi.org/10.1152/ajplegacy.1943.139.3.477MoreSectionsPDF (674 KB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInWeChat Previous Back to Top Download PDF FiguresReferencesRelatedInformation Cited BySynergism of coupled subsarcolemmal Ca2+ clocks and sarcolemmal voltage clocks confers robust and flexible pacemaker function in a novel pacemaker cell modelVictor A. Maltsev and Edward G. Lakatta1 March 2009 | American Journal of Physiology-Heart and Circulatory Physiology, Vol. 296, No. 3 More from this issue > Volume 139Issue 3July 1943Pages 477-480 Copyright & PermissionsCopyright © 1943 by American Physiological Societyhttps://doi.org/10.1152/ajplegacy.1943.139.3.477History Received 25 March 1943 Published online 1 July 1943 Published in print 1 July 1943 Metrics