Mechanisms of Make and Break Excitation Revisited: Paradoxical Break Excitation During Diastolic Stimulation

American Journal of Physiology. Heart and Circulatory Physiology. Feb, 2002  |  Pubmed ID: 11788404

Onset and termination of electric stimulation may result in "make" and "break" excitation of the heart tissue. Wikswo et al. (30) explained both types of stimulations by virtual electrode polarization. Make excitation propagates from depolarized regions (virtual cathodes). Break excitation propagates from hyperpolarized regions (virtual anodes). However, these studies were limited to strong stimulus intensities. We examined excitation during weak near-threshold diastolic stimulation. We optically mapped electrical activity from a 4 x 4-mm area of epicardium of Langendorff-perfused rabbit hearts (n = 12) around the pacing electrode in the presence (n = 12) and absence (n = 2) of 15 mM 2,3-butanedione monoxime. Anodal and cathodal 2-ms stimuli of various intensities were applied. We imaged an excitation wavefront with 528-micros resolution. We found that strong stimuli (x5 threshold) result in make excitation, starting from the virtual cathodes. In contrast, near-threshold stimulation resulted in break excitation, originating from the virtual anodes. Characteristic biphasic upstrokes in the virtual cathode area were observed. Break and make excitation represent two extreme cases of near-threshold and far-above-threshold stimulations, respectively. Both mechanisms are likely to contribute during intermediate clinically relevant strengths.

The Pinwheel Experiment Re-revisited

Journal of Theoretical Biology. Jan, 2002  |  Pubmed ID: 11812169

Virtual electrode induced phase singularity hypothesis explains the origin of cardiac arrhythmias caused by artificial electrical induction of rotors, i.e. vortex-like self-sustained sources of activity. This mechanism is thought to underlie both stimulus-induced arrhythmias and shock defibrillation therapy. In this paper, we extend this hypothesis to three dimensions using the bidomain model of cardiac tissue. We predict that virtual electrode polarization can produce three topologically distinct types of rotors anchored to: (1) transmural I-shaped scroll wave filaments; (2) near-surface U-shaped scroll wave filaments; and (3) intramural O-shaped scroll wave filaments.

Imaging of the Atrioventricular Node Using Optical Coherence Tomography

Journal of Cardiovascular Electrophysiology. Jan, 2002  |  Pubmed ID: 11843494

Phase I and Phase II of Short-term Mechanical Restitution in Perfused Rat Left Ventricles

American Journal of Physiology. Heart and Circulatory Physiology. Apr, 2002  |  Pubmed ID: 11893566

We examined the contributions of the Ca(2+) channels of the sarcolemma and of the sarcoplasmic reticulum to electromechanical restitution. Extrasystoles (F(1)) were interpolated 40-600 ms following a steady-state beat (F(0)) in perfused rat ventricles paced at 2 or 3 Hz. Plots of F(1)/F(0) versus the extrasystolic interval consisted of phase I, which occurred before relaxation of the steady-state beat, and phase II, which occurred later. Phase I exhibited a period of enhanced left ventricular pressure development that coincided with action potential prolongation. Phase I was eliminated by -BAY K 8644 (100 nM) and FPL 64176 (150 nM), augmented by 3 microM thapsigargin plus 200 nM ryanodine and unaffected by KN-93 and KB-R7943. Phase II was accelerated by the Ca(2+) channel agonists and by isoproterenol but was eliminated by thapsigargin plus ryanodine. The results suggest that phase I of electromechanical restitution is caused by a transient L-type Ca(2+) current facilitation, whereas phase II represents the recovery of the ability of the sarcoplasmic reticulum to release Ca(2+).

Mechanical Alternans and Restitution in Failing SHHF Rat Left Ventricles

American Journal of Physiology. Heart and Circulatory Physiology. Apr, 2002  |  Pubmed ID: 11893567

We examined mechanical alternans and electromechanical restitution in normal and failing rat hearts. Alternans occurred at 5 Hz in failing versus 9 Hz in control hearts and was reversed by 300 nM isoproterenol, 6 mM extracellular Ca(2+), 300 nM -BAY K 8644, or 50 nM ryanodine. Restitution curves comprised phase I, which was completed before relaxation of the steady-state beat, and phase II, which occurred later. Phase I action potential area and developed pressure ratios were significantly reduced in the failing versus control hearts. Phase II was a monoexponential increase in relative developed pressure as the extrasystolic interval was increased. The plateau of phase II was significantly elevated in failing hearts. Thapsigargin (3 microM) plus ryanodine (200 nM) potentiated phase I to a significantly greater extent in control versus failing hearts and abolished phase II in both groups. The results suggest that both regulation of Ca(2+) influx across the sarcolemma and Ca(2+) release by the sarcoplasmic reticulum may contribute to altered excitation-contraction coupling in the failing spontaneously hypertensive heart failure prone rat heart.

Mechanisms of Shock-induced Arrhythmogenesis During Acute Global Ischemia

American Journal of Physiology. Heart and Circulatory Physiology. Jun, 2002  |  Pubmed ID: 12003822

Little is known about the mechanisms of vulnerability and defibrillation under ischemic conditions. We investigated these mechanisms in 18 Langendorff-perfused rabbit hearts during 75% reduced-flow ischemia. Electrical activity was optically mapped from the anterior epicardium during right ventricular shocks applied at various phases of the cardiac cycle while the excitation-contraction decoupler 2,3-butanedione monoxime (BDM; 15 mM) was used to suppress motion artifacts caused by contraction of the heart. During ischemia, vulnerable window width increased [from 30-90% of the action potential duration (APD) in the control to -10 to 100% of the APD in ischemia]. Moreover, arrhythmia severity increased along with the reduction of APD (176 +/- 9 ms in control and 129 +/- 26 ms in ischemia, P < 0.01) and increased dispersion of repolarization (45 +/- 17 ms in control and 73 +/- 28 ms in ischemia, P < 0.01). Shock-induced virtual electrode polarization was preserved. Depolarizing (contrary to hyperpolarizing) response time constants increased. Virtual electrode-induced wavefronts of excitation had much more tortuous pathways leading to wavefront fractionation. Defibrillation failure at all shock strengths was observed in four hearts. Optical mapping revealed that the shock extinguished the arrhythmia; however, the arrhythmia self-originated after an isoelectric window of 339 +/- 189 ms. In conclusion, in most cases, virtual electrode-induced phase singularity (VEIPS) was responsible for shock-induced arrhythmogenesis during acute global ischemia. Enhancement of arrhythmogenesis was associated with an increased dispersion of repolarization and altered deexcitation. In four hearts, arrhythmogenesis could not be explained by VEIPS.

Virtual Electrodes in Virtual Reality of Defibrillation

Journal of Cardiovascular Electrophysiology. Jul, 2002  |  Pubmed ID: 12139291

Anode-break Excitation During End-diastolic Stimulation is Explained by Half-cell Double Layer Discharge

IEEE Transactions on Bio-medical Engineering. Oct, 2002  |  Pubmed ID: 12374349

The phenomenon of anodal-break excitation during end-diastolic stimulation of the heart was discovered many years ago by B. Hoffman. Yet, the existence and mechanistic explanation of this effect remain controversial. We sought to confirm its existence and to determine a possible role of half-cell potential. We used isolated Langendorff-perfused rabbit hearts (n = 6) which were stained with di-4-ANEPPS and perfused with 15-mM butanedione monoxime (BDM). Transmembrane potentials were optically recorded at the left ventricular epicardium with a high spatial and temporal resolution (200 microm/343 micros) near the tip of a 120-microm platinum-iridium Teflon-coated unipolar pacing electrode to detect virtual electrode polarization and to reconstruct an activation pattern. Hearts were paced at a cycle length of 300 ms by anodal square pulses with an amplitude of 0.1-10 mA and a duration of 5-60 ms. Data revealed that the anodal-break excitation does exists and is accompanied by an overshoot in the recordings of the pacing current. Addition of a diode in the stimulation circuit eliminated both the overshoot and the break excitation. The findings suggest that a half-cell surface potential at the pacing electrode metal-saline interface may influence the pacing currents during unipolar anodal cardiac stimulation providing "break"-like activation. We also confirmed that the threshold of "break"-like excitation is lower than make-excitation. We suggest that further exploration of this effect is needed in order to design improved multiphasic pacing waveforms.

Cx43 and Dual-pathway Electrophysiology of the Atrioventricular Node and Atrioventricular Nodal Reentry

Circulation Research. Mar, 2003  |  Pubmed ID: 12600895

Fluorescent imaging has revealed that posterior nodal extensions provide the anatomical substrate for the dual-pathway electrophysiology of the atrioventricular (AV) node during normal conduction and reentry. The reentry can be intranodal, or as well as the posterior nodal extensions, it can involve an endocardial layer of atrial/atrial-nodal (A/AN) cells as part of the AV nodal reentry (AVNR) circuit. Using fluorescent imaging with a voltage-sensitive dye and immunolabeling of Cx43, we mapped the electrical activity and structural substrate in 3 types of AVNR induced by premature atrial stimulation in 8 rabbit hearts. In 6 cases, the AVNR pathway involved (1) a fast pathway (FP), (2) the A/AN layer, and (3) a slow pathway (SP). In 4 cases, reentry took the path (1) SP, (2) A/AN layer, and (3) FP. In 2 cases, reentry was intranodal, propagating between the 2 posterior nodal extensions. Immunolabeling revealed that the FP and SP are formed by Cx43-expressing bundles surrounded by tissue without Cx43. Cx43-expressing posterior nodal extensions are the substrate of AVNR during both intranodal and extranodal reentry.

Optical Mapping Technique Applied to Biventricular Pacing: Potential Mechanisms of Ventricular Arrhythmias Occurrence

Pacing and Clinical Electrophysiology : PACE. Jan, 2003  |  Pubmed ID: 12687812

Although it has been suggested that multisite ventricular pacing alleviates heart failure by restoring ventricular electrical synchronization, the respective roles of voltage output, interventricular delay, and pacing sites in the development of ventricular arrhythmias occurrence have not been studied during biventricular pacing or LV pacing. Voltage-sensitive dye was used in eight ischemic Langerdorff-perfused guinea pig hearts to measure ventricular activation times and examine conduction patterns during multisite pacing from three RV and four LV sites. The hearts were stained with di-4-ANEPPS and mapped with a 16 x 16 photodiode array at a resolution of 625 microns per diode. Isochronal maps of RV and LV activation were plotted. Ischemia was produced by gradually halving the perfusion output over 5 minutes. Pacing the RV apex and the base of the LV anterior wall was associated with the most homogeneous and rapid activation pattern (28 +/- 9 vs 41 +/- 12 ms with the other configurations, P < 0.01), and no inducible arrhythmia. In six hearts, ventricular tachycardia could be induced when pacing from the right and left free walls with 20 ms of interventricular delay, at six times the pacing threshold output. In four hearts, simultaneous RV and LV pacing at high voltage output induced ventricular fibrillation with complex three-dimensional propagation patterns, independently of the pacing sites. During biventricular pacing with ischemia, pacing at high voltage output with a long interventricular delay is likely to induce ventricular arrhythmias, particularly when left and right pacing results in a conduction pattern orthogonal to the ventricular myocardial fibers orientation.

Nonlinear Effects in Subthreshold Virtual Electrode Polarization

American Journal of Physiology. Heart and Circulatory Physiology. Jun, 2003  |  Pubmed ID: 12742834

Introduction of the virtual electrode polarization (VEP) theory suggested solutions to several century-old puzzles of heart electrophysiology including explanation of the mechanisms of stimulation and defibrillation. Bidomain theory predicts that VEPs should exist at any stimulus strength. Although the presence of VEPs for strong suprathreshold pulses has been well documented, their existence at subthreshold strengths during diastole remains controversial. We studied cardiac membrane polarization produced by subthreshold stimuli in 1) rabbit ventricular muscle using high-resolution fluorescent imaging with the voltage-sensitive dye pyridinium 4-[2-[6-(dibutylamino)-2-naphthalenyl]-ethenyl]-1-(3-sulfopropyl)hydroxide (di-4-ANEPPS) and 2) an active bidomain model with Luo-Rudy ion channel kinetics. Both in vitro and in numero models show that the common dog-bone-shaped VEP is present at any stimulus strength during both systole and diastole. Diastolic subthreshold VEPs exhibited nonlinear properties that were expressed in time-dependent asymmetric reversal of membrane polarization with respect to stimulus polarity. The bidomain model reveals that this asymmetry is due to nonlinear properties of the inward rectifier potassium current. Our results suggest that active ion channel kinetics modulate the transmembrane polarization pattern that is predicted by the linear bidomain model of cardiac syncytium.

Fibrillation or Neurillation: Back to the Future in Our Concepts of Sudden Cardiac Death?

Circulation Research. May, 2003  |  Pubmed ID: 12775654

Effects of Lidocaine on Shock-induced Vulnerability

Journal of Cardiovascular Electrophysiology. Oct, 2003  |  Pubmed ID: 14760929

Lidocaine is known to increase the defibrillation threshold (DFT) of monophasic shocks (MS) and have no effect on DFT of biphasic shocks (BS). The aim of this study was to enhance our understanding of the mechanisms of vulnerability and defibrillation through the investigation of this difference.

Site of Origin and Molecular Substrate of Atrioventricular Junctional Rhythm in the Rabbit Heart

Circulation Research. Nov, 2003  |  Pubmed ID: 14563715

During failure of the sinoatrial node, the heart can be driven by an atrioventricular (AV) junctional pacemaker. The position of the leading pacemaker site during AV junctional rhythm is debated. In this study, we present evidence from high-resolution fluorescent imaging of electrical activity in rabbit isolated atrioventricular node (AVN) preparations that, in the majority of cases (11 out of 14), the AV junctional rhythm originates in the region extending from the AVN toward the coronary sinus along the tricuspid valve (posterior nodal extension, PNE). Histological and immunohistochemical investigation showed that the PNE has the same morphology and unique pattern of expression of neurofilament160 (NF160) and connexins (Cx40, Cx43, and Cx45) as the AVN itself. Block of the pacemaker current, If, by 2 mmol/L Cs+ increased the AV junctional rhythm cycle length from 611+/-84 to 949+/-120 ms (mean+/-SD, n=6, P<0.001). Immunohistochemical investigation showed that the principal If channel protein, HCN4, is abundant in the PNE. As well as the AV junctional rhythm, the PNE described in this study may also be involved in the slow pathway of conduction into the AVN as well as AVN reentry, and the predominant lack of expression of Cx43 as well as the presence of Cx45 in the PNE shown could help explain its slow conduction.

Diastolic Shocking Experience: Do Virtual Electrodes Exist Only During Systole?

Journal of Cardiovascular Electrophysiology. Nov, 2003  |  Pubmed ID: 14678139

Virtual Electrode Theory Explains Pacing Threshold Increase Caused by Cardiac Tissue Damage

American Journal of Physiology. Heart and Circulatory Physiology. Jun, 2004  |  Pubmed ID: 14726298

The virtual electrode polarization (VEP) effect is believed to play a key role in electrical stimulation of heart muscle. However, under certain conditions, including clinically, its existence and importance remain unknown. We investigated the influence of acute tissue damage produced by continuous pacing with strong current (40-mA, 4-ms biphasic pulses with 4-Hz frequency for 5 min) on stimulus-generated VEPs and pacing thresholds. A fluorescent optical mapping technique was used to obtain stimulus-induced transmembrane potential distribution around a pacing electrode applied to the ventricular surface of a Langendorff-perfused rabbit heart (n = 5). Maps and pacing thresholds were recorded before and after tissue damage. Spatial extents of electroporation and cell uncoupling were assessed by propidium iodide (n = 2) and connexin43 (n = 3) antibody staining, respectively. On the basis of these data, passive and active three-dimensional bidomain models were built to determine VEP patterns and thresholds for different-sized areas of the damaged region. Electrophysiological results showed that acute tissue damage led to disappearance of the VEP with an associated significant increase in pacing thresholds. Damage was expressed in electroporation and cell uncoupling within an approximately 1.0-mm-diameter area around the tip of the electrode. According to computer simulations, cell uncoupling, rather than electroporation, might be the direct cause of VEP elimination and threshold increase, which was nonlinearly dependent on the size of the damaged region. Fiber rotation with depth did not substantially affect the numerical results. The study explains failure to stimulate damaged tissue within the concepts of the VEP theory.

Optical Imaging of the Heart

Circulation Research. Jul, 2004  |  Pubmed ID: 15242982

Optical techniques have revolutionized the investigation of cardiac cellular physiology and advanced our understanding of basic mechanisms of electrical activity, calcium homeostasis, and metabolism. Although optical methods are widely accepted and have been at the forefront of scientific discoveries, they have been primarily applied at cellular and subcellular levels and considerably less to whole heart organ physiology. Numerous technical difficulties had to be overcome to dynamically map physiological processes in intact hearts by optical methods. Problems of contraction artifacts, cellular heterogeneities, spatial and temporal resolution, limitations of surface images, depth-of-field, and need for large fields of view (ranging from 2x2 mm2 to 3x3 cm2) have all led to the development of new devices and optical probes to monitor physiological parameters in intact hearts. This review aims to provide a critical overview of current approaches, their contributions to the field of cardiac electrophysiology, and future directions of various optical imaging modalities as applied to cardiac physiology at organ and tissue levels.

Structure-function Relationship in the AV Junction

The Anatomical Record. Part A, Discoveries in Molecular, Cellular, and Evolutionary Biology. Oct, 2004  |  Pubmed ID: 15368340

In the normal heart, the atrioventricular node (AVN) is part of the sole pathway between the atria and ventricles. Under normal physiological conditions, the AVN controls appropriate frequency-dependent delay of contractions. The AVN also plays an important role in pathology: it protects ventricles during atrial tachyarrhythmia, and during sinoatrial node failure an AV junctional pacemaker can drive the heart. Finally, the AV junction provides an anatomical substrate for reentry. Using fluorescent imaging with voltage-sensitive dyes and immunohistochemistry, we have investigated the structure-function relationship of the AV junction during normal conduction, reentry, and junctional rhythm. We identified molecular and structural heterogeneity that provides a substrate for the dual-pathway AVN conduction. We observed heterogeneity of expression of three isoforms of connexins: Cx43, Cx45, and Cx40. We identified the site of origin of junctional rhythm at the posterior extension of the AV node in 79% (n = 14) of the studied hearts. This structure was similar to the compact AV node as determined by morphologic and molecular investigations. In particular, both the posterior extension and the compact node express the pacemaking channel HCN4 (responsible for the I(F) current) and neurofilament 160. In the rabbit heart, AV junction conduction, reentrant arrhythmia, and spontaneous rhythm are governed by heterogeneity of expression of several isoforms of gap junctions and ion channels. Uniform neurofilament expression suggests that AV nodal posterior extensions are an integral part of the cardiac pacemaking and conduction system. On the other hand, differential expression of Cx isoforms in this region provides an explanation of longitudinal dissociation, dual-pathway electrophysiology, and AV nodal reentrant arrhythmogenesis.

Dynamics of Virtual Electrode-induced Scroll-wave Reentry in a 3D Bidomain Model

American Journal of Physiology. Heart and Circulatory Physiology. Oct, 2004  |  Pubmed ID: 15371264

Functional reentry in the heart can be caused by a wave front of excitation rotating around its edge. Previous simulations on the basis of monodomain cable equations predicted the existence of self-sustained, vortex-like wave fronts (scroll waves) rotating around a filament in three dimensions. In our simulations, we used the more accurate bidomain model with modified Beeler-Reuter ionic kinetics to study the dynamics of scroll-wave filaments in a 16 x 8 x 1.5-mm slab of ventricular tissue with straight fibers. Wave fronts were identified as the areas with inward current. Their edges represented the filaments. Both transmural and intramural reentries with I- and U-shaped filaments, respectively, were obtained by the S1-S2 point stimulation protocol through the virtual electrode-induced phase singularity mechanism. The filaments meandered along elongated trajectories and tended to attach to the tissue boundaries exposed to air (no current flow) rather than to the bath (zero extracellular potential). They completely detached from electroporated (zero transmembrane potential) boundaries. In our simulations, the presence of the bath led to generation of only U-shaped filaments, which survived for the 1.5-mm-thick slab but not for the slabs of 0.5- or 3-mm thicknesses. Thus boundary conditions may be another determinant of the type and dynamics of reentry.

Functional Imaging of the Embryonic Pacemaking and Cardiac Conduction System over the Past 150 Years: Technologies to Overcome the Challenges

The Anatomical Record. Part A, Discoveries in Molecular, Cellular, and Evolutionary Biology. Oct, 2004  |  Pubmed ID: 15372434

Early analyses of cardiac pacemaking and conduction system (CPCS) development relied on classic histology and visual inspection of the beating heart. Current techniques that facilitate delineation of the CPCS include the use of specific antibody markers and transgenic mouse lines specifically expressing reporter genes. Assaying the function of tiny embryonic hearts required an increase in the level of spatial and temporal resolution. Current methods for such analyses include the use of intracellular and extracellular microelectrodes, echocardiography, rapid optical imaging using fluorescent dyes, and most recently optical coherence tomography. This review will focus on methods developed to investigate the functional emergence of the embryonic cardiac conduction system. Where appropriate, the methods used to delineate the anatomic pathways will also be discussed. The combination of techniques to capture both morphological and functional data from the CPCS will further improve with continued interdisciplinary collaboration. The Supplementary Material referred to in this article can be found at the Anatomical Record website (http://www.interscience.wiley.com/jpages/0003-276X/suppmat).

Shock-induced Arrhythmogenesis is Enhanced by 2,3-butanedione Monoxime Compared with Cytochalasin D

American Journal of Physiology. Heart and Circulatory Physiology. Jan, 2004  |  Pubmed ID: 12958029

Investigation of the mechanisms of arrhythmia genesis and maintenance has benefited from the use of optical mapping techniques that employ excitation-contraction uncouplers. We investigated the effects of the excitation-contraction uncouplers 2,3-butanedione monoxime (BDM) and cytochalasin D (Cyto D) on the induction and maintenance of arrhythmia by electric shocks. Electrical activity was optically mapped from anterior epicardium of rabbit hearts (n = 9) during shocks (-100 V, 8 ms) applied from a ventricular lead at various phases of action potential duration (APD). Restitution curves were obtained using S1-S2 protocol and measurement of APD values at 70% of repolarization. Compared with Cyto D, BDM significantly shortened APD at 90% of repolarization, although no significant difference in dispersion of repolarization was observed. Wavelength was also shortened with BDM. In general, shock-induced arrhythmias with BDM and Cyto D were ventricular tachycardic in nature. With respect to shock-induced sustained arrhythmias, the vulnerable window was wider and the incidence was higher with BDM than with Cyto D. There was also a difference in the morphology of ventricular tachycardia (VT) between the two agents. The arrhythmias with BDM usually resembled monomorphic VT, especially those that lasted >30 s. In contrast, arrhythmias with Cyto D more resembled polymorphic VT. However, the average number of phase singularities increased under Cyto D vs. BDM, whereas no significant difference in the dominant frequency of shock-induced sustained arrhythmia was observed. BDM reduced the slope of the restitution curve compared with Cyto D, but duration of arrhythmia under BDM was significantly increased compared with Cyto D. In conclusion, BDM increased arrhythmia genesis and maintenance relative to Cyto D.

The Gurvich Waveform Has Lower Defibrillation Threshold Than the Rectilinear Waveform and the Truncated Exponential Waveform in the Rabbit Heart

Canadian Journal of Physiology and Pharmacology. Feb, 2005  |  Pubmed ID: 15791288

Implantable cardioverter defibrillator studies have established the superiority of biphasic waveforms over monophasic waveforms. However, external defibrillator studies of biphasic waveforms are not as widespread. Our objective was to compare the defibrillation efficacy of clinically used biphasic waveforms, i.e., truncated exponential, rectilinear, and quasi-sinusoidal (Gurvich) waveforms in a fibrillating heart model. Langendorff-perfused rabbit hearts (n = 10) were stained with a voltage-sensitive fluorescent dye, Di-4-ANEPPS. Transmembrane action potentials were optically mapped from the anterior epicardium. We found that the Gurvich waveform was significantly superior (p < 0.05) to the rectilinear and truncated exponential waveforms. The defibrillation thresholds (mean +/- SE) were as follows: Gurvich, 0.25 +/- 0.01 J; rectilinear-1, 0.34 +/- 0.01 J; rectilinear-2, 0.33 +/- 0.01 J; and truncated exponential, 0.32 +/- 0.02 J. Using optically recorded transmembrane responses, we determined the shock-response transfer function, which allowed us to predict the cellular response to waveforms at high accuracy. The passive parallel resistor-capacitor model (RC-model) predicted polarization superiority of the Gurvich waveform in the myocardium with a membrane time constant (taum) of less than 2 ms. The finding of a lower defibrillation threshold with the Gurvich waveform in an in vitro model of external defibrillation suggests that the Gurvich waveform may be important for future external defibrillator designs.

Mechanisms of Superiority of Ascending Ramp Waveforms: New Insights into Mechanisms of Shock-induced Vulnerability and Defibrillation

American Journal of Physiology. Heart and Circulatory Physiology. Aug, 2005  |  Pubmed ID: 15792989

Monophasic ascending ramp (AR) and descending ramp (DR) waveforms are known to have significantly different defibrillation thresholds. We hypothesized that this difference arises due to differences in mechanisms of arrhythmia induction for the two waveforms. Rabbit hearts (n = 10) were Langendorff perfused, and AR and DR waveforms (7, 20, and 40 ms) were randomly delivered from two line electrodes placed 10 mm apart on the anterior ventricular epicardium. We optically mapped cellular responses to shocks of various strengths (5, 10, and 20 V/cm) and coupling intervals (CIs; 120, 180, and 300 ms). Optical mapping revealed that maximum virtual electrode polarization (VEP) was reached at significantly different times for AR and DR of the same duration (P < 0.05) for all tested CIs. As a result, VEP for AR were stronger than for DR at the end of the shock. Postshock break excitation resulting from AR generated faster propagation and typically could not form reentry. In contrast, partially dissipated VEP resulting from DR generated slower propagation; the wavefront was able to propagate into deexcited tissue and thus formed a shock-induced reentry circuit. Therefore, for the same delivered energy, AR was less proarrhythmic compared with DR. An active bidomain model was used to confirm the electrophysiological results. The VEP hypothesis explains differences in vulnerability associated with monophasic AR and DR waveforms and, by extension, the superior defibrillation efficacy of the AR waveform compared with the DR waveform.

Optical Coherence Tomography Imaging of the Purkinje Network

Journal of Cardiovascular Electrophysiology. May, 2005  |  Pubmed ID: 15877631

Mechanisms of Enhanced Shock-induced Arrhythmogenesis in the Rabbit Heart with Healed Myocardial Infarction

American Journal of Physiology. Heart and Circulatory Physiology. Sep, 2005  |  Pubmed ID: 15879480

Shock-induced vulnerability and defibrillation have been mostly studied in structurally normal hearts. However, defibrillation therapy is normally applied to patients with diseased hearts, frequently those with prior myocardial infarction (MI). Shock-induced vulnerability and defibrillation have not been well studied under this condition. We sought to examine the mechanisms of shock-induced arrhythmogenesis and arrhythmia maintenance in a rabbit model of healed MI (4 wk or more postinfarction). Ligation of the lateral division or posterolateral division of the left coronary artery at a level of 40-70% from the apex was performed 53 +/- 21 days before acute experiments. Shock-induced vulnerability was assessed in infarcted (n = 8) and structurally normal (n = 8) hearts by delivering internal monophasic shocks at different shock strengths and delivery phases. Electrical activities from the anterior epicardium during shock application and during shock-induced arrhythmias were optically recorded and quantitatively analyzed. Ligation resulted in a transmural left ventricular free wall infarction mainly located at the apical region with a consistent endocardial border zone (BZ) as confirmed by histological studies. There were significant increases in the incidence, severity, and duration of shock-induced arrhythmias in the infarcted hearts versus controls due to 1) postshock break-excitation wavefronts that frequently originated near the infarction BZ and 2) the existence of an infarction BZ that created an anatomic reentry pathway and facilitated arrhythmia maintenance. In conclusion, the infarction BZ contributes to both increased shock-induced arrhythmogenesis and arrhythmia maintenance in the rabbit model of healed MI.

Differences Between Left and Right Ventricular Chamber Geometry Affect Cardiac Vulnerability to Electric Shocks

Circulation Research. Jul, 2005  |  Pubmed ID: 15976315

Although effects of shock strength and waveform on cardiac vulnerability to electric shocks have been extensively documented, the contribution of ventricular anatomy to shock-induced polarization and postshock propagation and thus, to shock outcome, has never been quantified; this is caused by lack of experimental methodology capable of mapping 3-D electrical activity. The goal of this study was to use optical imaging experiments and 3-D bidomain simulations to investigate the role of structural differences between left and right ventricles in vulnerability to electric shocks in rabbit hearts. The ventricles were paced apically, and uniform-field, truncated-exponential, monophasic shocks of reversed polarity were applied over a range of coupling intervals (CIs) in experiment and model. Experiments and simulations revealed that reversing the direction of externally-applied field (RV- or LV- shocks) alters the shape of the vulnerability area (VA), the 2-D grid encompassing episodes of arrhythmia induction. For RV- shocks, VA was nearly rectangular indicating little dependence of postshock arrhythmogenesis on CI. For LV- shocks, the probability of arrhythmia induction was higher for longer than for shorter CIs. The 3-D simulations demonstrated that these effects stem from the fact that reversal of field direction results in relocation of the main postshock excitable area from LV wall (RV- shocks) to septum (LV- shocks). Furthermore, the effect of septal (but not LV) excitable area in postshock propagation was found to strongly depend on preshock state. Knowledge regarding the location of the main postshock excitable area within the 3-D ventricular volume could be important for improving defibrillation efficacy.

Chessboard of Atrial Fibrillation: Reentry or Focus? Single or Multiple Source(s)? Neurogenic or Myogenic?

American Journal of Physiology. Heart and Circulatory Physiology. Sep, 2005  |  Pubmed ID: 16100254

Electroporation of the Heart

Europace : European Pacing, Arrhythmias, and Cardiac Electrophysiology : Journal of the Working Groups on Cardiac Pacing, Arrhythmias, and Cardiac Cellular Electrophysiology of the European Society of Cardiology. Sep, 2005  |  Pubmed ID: 16102512

Defibrillation shocks are commonly used to terminate life-threatening arrhythmias. According to the excitation theory of defibrillation, such shocks are aimed at depolarizing the membranes of most cardiac cells resulting in resynchronization of electrical activity in the heart. If shock-induced changes in transmembrane potential are large enough, they can cause transient tissue damage due to electroporation. In this review evidence is presented that (a) electroporation of the heart tissue can occur during clinically relevant intensities of the external electrical field, and (b) electroporation can affect the outcome of defibrillation therapy; being both pro- and anti-arrhythmic.

Hibernator Citellus Undulatus Maintains Safe Cardiac Conduction and is Protected Against Tachyarrhythmias During Extreme Hypothermia: Possible Role of Cx43 and Cx45 Up-regulation

Heart Rhythm : the Official Journal of the Heart Rhythm Society. Sep, 2005  |  Pubmed ID: 16171752

Most mammals experience cardiac arrest during hypothermia. In contrast, hibernators remain in sinus rhythm even at body temperatures of 0 degrees C.

Optical Mapping of the Atrioventricular Junction

Journal of Electrocardiology. Oct, 2005  |  Pubmed ID: 16226086

In the normal heart, the atrioventricular node (AVN) is part of the sole pathway between the atria and ventricles, and is responsible for the appropriate atrial-ventricular delay. Under normal physiological conditions, the AVN controls appropriate frequency-dependent delay of contractions. The AVN also plays an important role in pathology: it protects ventricles during atrial tachyarrhythmia, and during sinoatrial node failure the atrioventricular (AV) junction assumes the role of pacemaker. Finally, the AV junction provides an anatomic substrate for AV nodal reentrant tachycardia, which is the most prevalent supraventricular tachycardia in humans. Using fluorescent imaging with voltage-sensitive dye and immunohistochemistry, we have investigated the structure-function relationship of the atrioventricular (AV) junction during normal conduction, reentry, and junctional rhythm. We identified the site of origin of junctional rhythm at the posterior extension of the AV node (AVN) in 78% (n=23) of the studied hearts and we found that this pacemaker is sensitive to autonomic control. For instance, when the autonomic nervous system was activated using subthreshold stimulation, a transient accelerated junctional rhythm was observed when subthreshold stimulation was terminated. A very similar phenomenon is observed clinically during slow pathway ablations treating AV nodal reentrant tachycardia (AVNRT). The autonomic control of the AV junction was investigated using immunohistochemistry, showing that the AV junction of the rabbit is very densely innervated with both cholinergic and adrenergic neurons. The posterior AV nodal extension was similar to the compact AVN as determined by morphologic and molecular investigations. In particular, both the posterior extension and the compact node express the pacemaking channel HCN4 (responsible for the IF current) and neurofilament 160. In the rabbit heart, AV junction conduction, reentrant arrhythmia, and spontaneous rhythm are governed by heterogeneity of expression of several isoforms of gap junctions and ion channels, and these properties are regulated by the autonomic nervous system. Uniform neurofilament expression suggests that AV nodal posterior extensions are an integral part of the cardiac pacemaking and conduction system.

Fluorescence Imaging for Real-time Monitoring of High-intensity Focused Ultrasound Cardiac Ablation

Annals of Biomedical Engineering. Oct, 2005  |  Pubmed ID: 16240084

Side effects and limitations of radio-frequency ablation of cardiac arrhythmias prompted search for alternative energy sources and means of their application. High-intensity focused ultrasound (HIFU) is becoming an increasingly attractive modality for ablation because of its unique ability for non-invasive or minimally invasive, non-contact focal ablation in 3D volume without affecting intervening and surrounding cells. The purpose of this study is to develop a real-time monitoring technique to elucidate HIFU-induced modifications of electrical conduction in cardiac tissues and to investigate the HIFU cardiac ablation process to help to achieve optimal HIFU ablation outcome. We conducted experimental studies applying HIFU at 4.23 MHz to ablate the atrio-ventricular (AV) node and ventricular tissue of Langendorff-perfused rabbit hearts. We employed fluorescent voltage-sensitive dye imaging and surface electrodes to monitor the electrical conduction activity induced by HIFU application in real time. In ventricular epicardium HIFU ablation, fluorescent imaging revealed gradual reduction of the plateau phase and amplitude of the action potential. Subsequently, conduction block and cell death were observed at the site of ablation. When HIFU was applied to the AV node, fluorescent imaging and electrograms revealed the development of the AV block. The study establishes that real-time fluorescent imaging provides novel monitoring and assessment to study HIFU cardiac ablation, which may be able to provide improved understanding of HIFU cardiac ablation process and mechanism useful for development of successful clinical applications.

Three-dimensional Anatomy of the Conduction System of the Early Embryonic Rabbit Heart

The Anatomical Record. Part A, Discoveries in Molecular, Cellular, and Evolutionary Biology. Jan, 2006  |  Pubmed ID: 16287158

The complete embryonic cardiac conduction system is difficult to view in three dimensions, primarily because there has not been a marker of all segments of the normal system throughout all stages of development. Imaging of the conduction system components within the atria has been particularly controversial because different markers reveal different pathways that may or may not represent conduction system components. The conduction system of the adult and embryonic rabbit, however, can be labeled in its entirety with the neurofilament marker, NF-160. The conduction system of rabbit embryos at several stages of development spanning cardiac septation was therefore investigated. Optical mapping of the electrical signature of the conduction system previously revealed a close correlation between the cardiac activation patterns and the anatomy as shown by serial sections. The 3D relationship between the components of the conduction system could only be inferred from the 2D sections. The sections were consequently reconstructed using a commercial software program (AutoQuant). This is the first demonstration of the three-dimensional complete normal rabbit embryonic cardiac conduction system at several stages of development.

Tornado in a Dish: Revealing the Mechanisms of Ventricular Arrhythmias in Engineered Cardiac Tissues

Cardiovascular Research. Feb, 2006  |  Pubmed ID: 16325163

Precordial Thump and Commotio Cordis: the Yin and Yang of Mechanoelectric Feedback in the Heart

Heart Rhythm : the Official Journal of the Heart Rhythm Society. Feb, 2006  |  Pubmed ID: 16443534

Mechanisms of Unpinning and Termination of Ventricular Tachycardia

American Journal of Physiology. Heart and Circulatory Physiology. Jul, 2006  |  Pubmed ID: 16501014

High-energy defibrillation shock is the only therapy for ventricular tachyarrhythmias. However, because of adverse side effects, lowering defibrillation energy is desirable. We investigated mechanisms of unpinning, destabilization, and termination of ventricular tachycardia (VT) by low-energy shocks in isolated rabbit right ventricular preparations (n = 22). Stable VT was initiated with burst pacing and was optically mapped. Monophasic "unpinning" shocks (10 ms) of different strengths were applied at various phases throughout the reentry cycle. In 8 of 22 preparations, antitachycardia pacing (ATP: 8-20 pulses, 50-105% of period, 0.8-10 mA) was also applied. Termination of reentry by ATP was achieved in only 5 of 8 preparations. Termination by unpinning occurred in all 22 preparations. Rayleigh's test showed a statistically significant unpinning phase window, during which reentry could be unpinned and subsequently terminated with E80 (magnitude at which 80% of reentries were unpinned) = 1.2 V/cm. All reentries were unpinned with field strengths < or = 2.4 V/cm. Unpinning was achieved by inducing virtual electrode polarization and secondary sources of excitation at the core of reentry. Optical mapping revealed the mechanisms of phase-dependent unpinning of reentry. These results suggest that a 20-fold reduction in energy could be achieved compared with conventional high-energy defibrillation and that the unpinning method may be more effective than ATP for terminating stable, pinned reentry in this experimental model.

Postganglionic Nerve Stimulation Induces Temporal Inhibition of Excitability in Rabbit Sinoatrial Node

American Journal of Physiology. Heart and Circulatory Physiology. Aug, 2006  |  Pubmed ID: 16565321

Vagal stimulation results in complex changes of pacemaker excitability in the sinoatrial node (SAN). To investigate the vagal effects in the rabbit SAN, we used optical mapping, which is the only technology that allows resolving simultaneous changes in the activation pattern and action potentials morphologies. With the use of immunolabeling, we identified the SAN as a neurofilament 160-positive but connexin 43-negative region (n = 5). Normal excitation originated in the SAN center with a cycle length (CL) of 405 +/- 14 ms (n = 14), spread anisotropically along the crista terminalis (CT), and failed to conduct toward the septum. Postganglionic nerve stimulation (PNS, 400-800 ms) reduced CL by 74 +/- 7% transiently and shifted the leading pacemaker inferiorly (78%) or superiorly (22%) from the SAN center by 2-10 mm. In the intercaval region between the SAN center and the septal block zone, PNS produced an 8 +/- 1-mm(2) region of transient hyperpolarization and inexcitability. The first spontaneous or paced excitation following PNS could not enter this region for 500-1,500 ms. Immunolabeling revealed that the PNS-induced inexcitable region is located between the SAN center and the block zone and has a 2.5-fold higher density of choline acetyltransferase than CT but is threefold lower than the SAN center. The fact that the inexcitability region does not coincide with the most innervated area indicates that the properties of the myocytes themselves, as well as intercellular coupling, must play a role in the inexcitability induction. Optically mapping revealed that PNS resulted in transient loss of pacemaker cell excitability and unidirectional entrance conduction block in the periphery of SAN.

Gene Printer: Laser-scanning Targeted Transfection of Cultured Cardiac Neonatal Rat Cells

Cell Communication & Adhesion. Jul-Aug, 2006  |  Pubmed ID: 16916749

Heterogeneous gene expression in cardiac cells and tissues which requires targeted delivery of foreign DNA into selected cells or regions is needed for the development of novel therapies. Several techniques have been employed for targeted transfection, such as direct microinjection into cells or targeted electroporation. However, these techniques have limited bandwidth or spatial resolution of transfection. We aimed to develop a method for transfection of cardiac cells by means of laser-assisted optoporation using a standard confocal microscope. This technique allows for the transfection of selected cell types in the presence of other cell types as long as they are distinguishable with a microscope. This technique can work as a "gene printer" creating arbitrarily shaped areas of transfected cells.

Present Understanding of Shock Polarity for Internal Defibrillation: the Obvious and Non-obvious Clinical Implications

Pacing and Clinical Electrophysiology : PACE. Aug, 2006  |  Pubmed ID: 16923006

Uncertainty about the best electrode configuration has combined with the programming flexibility in modern implantable cardioverter-defibrillators (ICDs) to result in routine polarity reversal during an implant to deal with a high defibrillation threshold (DFT). We feel that this practice is not always supported by the clinical data and the present scientific understanding of defibrillation.

Localization of Na+ Channel Isoforms at the Atrioventricular Junction and Atrioventricular Node in the Rat

Circulation. Sep, 2006  |  Pubmed ID: 16966585

The electrical activity of the atrioventricular node (AVN) is functionally heterogeneous, but how this relates to distinct cell types and the 3-dimensional structure of the AVN is unknown. To address this, we have studied the expression of Na(V)1.5 and other Na+ channel isoforms in the AVN.

Connections, Connections, Connexins: Towards Systems Biology Paradigm of Cardiac Arrhythmia

Journal of Molecular and Cellular Cardiology. Dec, 2006  |  Pubmed ID: 17030039

Finite Element Modeling of Electric Field Effects of TASER Devices on Nerve and Muscle

Conference Proceedings : ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference. 2006  |  Pubmed ID: 17946454

TASERs deliver electrical pulses that can temporarily incapacitate subjects. The goal of this paper is to analyze the distribution of currents in muscle layers and understand the electro-muscular incapacitation safety and efficacy of TASERs. The analyses describe skeletal muscle and motor nerve activation, cell electroporation and current and electric field distributions through skin, fat and muscle layers, under worst-case assumptions for TASER electrode penetration and separation. For the muscle layer, the analysis predicts worst-case current-density and field-strength values of 94 mA/cm(2) and 47 V/cm. Both values are higher than thresholds required for neuromuscular activation but significantly lower than levels needed for permanent cellular electroporation or tissue damage. The results indicate that TASERs are safe and effective in producing temporary subject incapacitation.

The Role of Photon Scattering in Optical Signal Distortion During Arrhythmia and Defibrillation

Biophysical Journal. Nov, 2007  |  Pubmed ID: 17978166

Optical mapping of arrhythmias and defibrillation provides important insights; however, a limitation of the technique is signal distortion due to photon scattering. The goal of this experimental/simulation study is to investigate the role of three-dimensional photon scattering in optical signal distortion during ventricular tachycardia (VT) and defibrillation. A three-dimensional realistic bidomain rabbit ventricular model was combined with a model of photon transport. Shocks were applied via external electrodes to induce sustained VT, and transmembrane potentials (V(m)) were compared with synthesized optical signals (V(opt)). Fluorescent recordings were conducted in isolated rabbit hearts to validate simulation results. Results demonstrate that shock-induced membrane polarization magnitude is smaller in V(opt) and in experimental signals as compared to V(m). This is due to transduction of potentials from weakly polarized midmyocardium to the epicardium. During shock-induced reentry and in sustained VT, photon scattering, combined with complex wavefront dynamics, results in optical action potentials near a filament exhibiting i), elevated resting potential, ii), reduced amplitude relative to pacing, and iii), dual-humped morphologies. A shift of up to 4 mm in the phase singularity location was observed in V(opt) maps when compared to V(m). This combined experimental/simulation study provides an interpretation of optical recordings during VT and defibrillation.

Direct Measurements of Membrane Time Constant During Defibrillation Strength Shocks

Heart Rhythm : the Official Journal of the Heart Rhythm Society. Apr, 2007  |  Pubmed ID: 17399638

Defibrillation shocks impose significant energy demand on implantable cardioverter-defibrillators (ICDs). Several modeling studies have been devoted to optimizing shock parameters, and a large number of these studies treat the heart as a simplified lumped network. The time constant of membrane polarization (tau(m)) is a key variable for such modeling efforts.

Effects of Sterile Pericarditis on Connexins 40 and 43 in the Atria: Correlation with Abnormal Conduction and Atrial Arrhythmias

American Journal of Physiology. Heart and Circulatory Physiology. Aug, 2007  |  Pubmed ID: 17434983

The canine sterile pericarditis model is characterized by impaired conduction and atrial arrhythmia vulnerability. Electrical and structural remodeling processes caused by the inflammatory response likely promote these abnormalities. In the present study, we tested the hypothesis that altered distribution of atrial connexins is associated with markedly abnormal atrial conduction, thereby contributing to vulnerability to atrial flutter (AFL) and atrial fibrillation (AF) induction and maintenance. During rapid pacing and induced, sustained AFL or AF in five sterile pericarditis (SP) and five normal (NL) dogs, epicardial atrial electrograms were recorded simultaneously from both atria (380 electrodes) or from the right atrium (RA) and Bachmann's bundle (212 electrodes). Tissues from RA sites were subjected to immunostaining and immunoblotting to assess connexin (Cx) 40 and Cx43 distribution and expression. Transmural myocyte (alpha-actinin) and fibroblast (vimentin) volume were also assessed by immunostaining. RA pacing maps showed markedly abnormal conduction in SP, with uniform conduction in NL. Total RA activation time was significantly prolonged in SP vs. NL at 300-ms and 200-ms pacing-cycle lengths. Sustained arrhythmias were only inducible in SP [total: 4/5 (AFL: 3/5; AF: 1/5)]. In NL, Cx40, Cx43, alpha-actinin, and vimentin were homogeneously distributed transmurally. In SP, Cx40, Cx43, and alpha-actinin were absent epicardially, decreased midmyocardially, and normal endocardially. SP increased epicardial vimentin expression, suggesting fibroblast proliferation. Immunoblot analysis confirmed reduced expression of Cx40 and Cx43 in SP. The transmural gradient in the volume fraction of Cx40 and Cx43 in SP is associated with markedly abnormal atrial conduction and is likely an important factor in the vulnerability to induction and maintenance of AFL/AF in SP.

Application of Blebbistatin As an Excitation-contraction Uncoupler for Electrophysiologic Study of Rat and Rabbit Hearts

Heart Rhythm : the Official Journal of the Heart Rhythm Society. May, 2007  |  Pubmed ID: 17467631

Application of fluorescence imaging of cardiac electrical activity is limited by motion artifacts and/or side effects of currently available pharmacologic excitation-contraction uncoupling agents.

Atrioventricular Conduction with and Without AV Nodal Delay: Two Pathways to the Bundle of His in the Rabbit Heart

American Journal of Physiology. Heart and Circulatory Physiology. Aug, 2007  |  Pubmed ID: 17496219

The electrophysiological properties of atrioventricular (AV) nodal dual pathways have traditionally been investigated with premature stimuli delivered with right atrial pacing. However, little is known about the functional characteristics of AV nodal inputs outside of this context. Superfused rabbit triangle of Koch preparations (n = 8) and Langendorff-perfused hearts (n = 10) were paced throughout the triangle of Koch and mapped electrically and optically for activation pattern, electrogram and optical action potential morphologies, stimulation thresholds, and stimulus-His (S-H) intervals. Optical mapping and changes in His electrogram morphology were used to confirm the activation pathway. Pacing stimuli >or=2 mm above the tricuspid valve caused fast-pathway activation of the AV node and His with a threshold of 2.4 +/- 1.6 mA. An area directly below the coronary sinus had high thresholds (8.6 +/- 1.4 mA) that also resulted in fast-pathway excitation (P < 0.001). S-H intervals (81 +/- 19 ms) for fast-pathway activation remained constant throughout the triangle of Koch, reflecting the AV delay. Stimuli applied <2 mm from the tricuspid valve resulted in slow pathway (SP) excitation or direct His excitation (4.4 +/- 2.2 mA threshold; P < 0.001 compared with fast pathway). For SP/His pacing, S-H intervals showed a strong dependence on the distance from the His electrode and were significantly lower than S-H intervals for fast-pathway activation. SP/His pacing also displayed characteristic changes in His electrogram morphology. In conclusion, optical maps and S-H intervals for SP/His activation suggest that AV conduction via SP bypasses the compact AV node via the lower nodal bundle, which may be utilized to achieve long-term ventricular synchronization.

Innovation in Optical Imaging: Looking Inside the Heart

Heart Rhythm : the Official Journal of the Heart Rhythm Society. Jul, 2007  |  Pubmed ID: 17599679

Cloning, Sequence Analysis and Phylogeny of Connexin43 Isolated from American Black Bear Heart

DNA Sequence : the Journal of DNA Sequencing and Mapping. Oct, 2007  |  Pubmed ID: 17654014

Conduction in the heart requires gap junctions. In mammalian ventricular myocytes these consist of connexin43 (Cx43). Hearts of non-hibernating species display conduction disturbances at reduced temperatures. These may exacerbate into lethal arrhythmias. Hibernating species are protected against these arrhythmias by a non-resolved mechanism. To analyze whether the amino acid composition of Cx43 from the hibernating American black bear displays specific features, we cloned the full coding sequence of Ursus americanus Cx43 and compared with that of other (non)hibernating species. UaCx43 displays 99.7% identity to rabbit Cx43 at the amino acid level. No specific features were observed in UaCx43 when compared to previously cloned Cx43 from hibernating and non-hibernating mammals. Phylogenetic tree reconstruction of this and other published full-length Cx43 sequences reveals a very high level of conservation from fish to men. Finally, one of the previously identified six mammalian characteristic amino acids, is not conserved in the black bear.

Optical Coherence Tomography As a Tool for Measuring Morphogenetic Deformation of the Looping Heart

Anatomical Record (Hoboken, N.J. : 2007). Sep, 2007  |  Pubmed ID: 17721979

Optical coherence tomography (OCT) was used to investigate morphogenesis of the embryonic chick heart during the first phase of looping (c-looping), as the heart bends and twists into a c-shaped tube. The present study focuses on the morphomechanical effects of the splanchnopleure (SPL), a membrane that has been shown to play a major role in cardiac torsion by pressing against the ventral surface of the heart. Without the SPL, rightward torsion (rotation) is delayed. The images show that compressive forces exerted by the SPL alter the shapes of the heart tube and primitive atria, as well as their spatial relationships. The SPL normally holds the heart in the plane of the embryo and forces cardiac jelly (CJ) out of adjacent regions in the atria. When the SPL is removed, cross-sections become more circular, CJ is more uniformly distributed, and the heart displaces ventrally. In addition, OCT-based morphogenetic strain maps were measured during looping by tracking the three-dimensional motions of microspheres placed on the myocardium. The spatial-temporal patterns of the strains correlated well with the observed behavior of the heart, including delayed torsion that occurs in SPL-lacking embryos. These results illustrate the potential of OCT as a tool in studies of morphogenesis, as well as provide a better understanding of the mechanical forces that drive cardiac looping.

Three-dimensional Panoramic Imaging of Cardiac Arrhythmias in Rabbit Heart

Journal of Biomedical Optics. Jul-Aug, 2007  |  Pubmed ID: 17867823

Cardiac fluorescent optical imaging provides the unique opportunity to investigate the dynamics of propagating electrical waves during ventricular arrhythmias and the termination of arrhythmias by strong electric shocks. Panoramic imaging systems using charge-coupled device (CCD) cameras as the photodetector have been developed to overcome the inability to monitor electrical activity from the entire cardiac surface. Photodiode arrays (PDAs) are known to have higher temporal resolution and signal quality, but lower spatial resolution compared to CCD cameras. We construct a panoramic imaging system with three PDAs and image Langendorff perfused rabbit hearts (n=18) during normal sinus rhythm, epicardial pacing, and arrhythmias. The recorded spatiotemporal dynamics of electrical activity is texture mapped onto a reconstructed 3-D geometrical heart model specific to each heart studied. The PDA-based system provides sufficient spatial resolution (1.72 mm without interpolation) for the study of wavefront propagation in the rabbit heart. The reconstructed 3-D electrical activity provides us with a powerful tool to investigate the fundamental mechanisms of arrhythmia maintenance and termination.

Enhanced Transmural Fiber Rotation and Connexin 43 Heterogeneity Are Associated with an Increased Upper Limit of Vulnerability in a Transgenic Rabbit Model of Human Hypertrophic Cardiomyopathy

Circulation Research. Nov, 2007  |  Pubmed ID: 17885214

Human hypertrophic cardiomyopathy, characterized by cardiac hypertrophy and myocyte disarray, is the most common cause of sudden cardiac death in the young. Hypertrophic cardiomyopathy is often caused by mutations in sarcomeric genes. We sought to determine arrhythmia propensity and underlying mechanisms contributing to arrhythmia in a transgenic (TG) rabbit model (beta-myosin heavy chain-Q403) of human hypertrophic cardiomyopathy. Langendorff-perfused hearts from TG (n=6) and wild-type (WT) rabbits (n=6) were optically mapped. The upper and lower limits of vulnerability, action potential duration (APD) restitution, and conduction velocity were measured. The transmural fiber angle shift was determined using diffusion tensor MRI. The transmural distribution of connexin 43 was quantified with immunohistochemistry. The upper limit of vulnerability was significantly increased in TG versus WT hearts (13.3+/-2.1 versus 7.4+/-2.3 V/cm; P=3.2e(-5)), whereas the lower limits of vulnerability were similar. APD restitution, conduction velocities, and anisotropy were also similar. Left ventricular transmural fiber rotation was significantly higher in TG versus WT hearts (95.6+/-10.9 degrees versus 79.2+/-7.8 degrees; P=0.039). The connexin 43 density was significantly increased in the mid-myocardium of TG hearts compared with WT (5.46+/-2.44% versus 2.68+/-0.77%; P=0.024), and similar densities were observed in the endo- and epicardium. Because a nearly 2-fold increase in upper limit of vulnerability was observed in the TG hearts without significant changes in APD restitution, conduction velocity, or the anisotropy ratio, we conclude that structural remodeling may underlie the elevated upper limit of vulnerability in human hypertrophic cardiomyopathy.

Autonomic Control and Innervation of the Atrioventricular Junctional Pacemaker

Heart Rhythm : the Official Journal of the Heart Rhythm Society. Oct, 2007  |  Pubmed ID: 17905339

The main physiologic function of the AV junction is control of timing between atrial and ventricular excitation. However, under pathologic conditions, the AV junction may become the pacemaker of the heart. Unlike the well-characterized sinoatrial node (SAN), autonomic control of the AV junctional pacemaker has not been studied.

Atria Are More Susceptible to Electroporation Than Ventricles: Implications for Atrial Stunning, Shock-induced Arrhythmia and Defibrillation Failure

Heart Rhythm : the Official Journal of the Heart Rhythm Society. Apr, 2008  |  Pubmed ID: 18362029

Defibrillation shock is known to induce atrial stunning, which is electrical and mechanical dysfunction.

Nature Versus Nurture in Cardiac Conduction: Toward Integrative Paradigm of Cardiac Tissue Engineering

Circulation Research. Jul, 2008  |  Pubmed ID: 18635826

Electrophysiological Mechanisms of Antiarrhythmic Protection During Hypothermia in Winter Hibernating Versus Nonhibernating Mammals

Heart Rhythm : the Official Journal of the Heart Rhythm Society. Nov, 2008  |  Pubmed ID: 18984537

Robust cell-to-cell coupling is critically important in the safety of cardiac conduction and protection against ventricular fibrillation (VF). Hibernating mammals have evolved naturally protective mechanisms against VF induced by hypothermia and reperfusion injury.

Connexin 43 Expression Delineates Two Discrete Pathways in the Human Atrioventricular Junction

Anatomical Record (Hoboken, N.J. : 2007). Feb, 2008  |  Pubmed ID: 18085635

Gap junction expression has been studied in the atrioventricular junction (AVJ) of many species, however, their distribution in the human AVJ is unknown. The AVJ expression of the gap junction protein connexin 43 (Cx43) is species dependent; therefore we investigated its distribution in the human AVJ. Using Masson trichrome histology, we reconstructed the AVJ of three normal human hearts and one with dilated cardiomyopathy in three dimensions. Cx43 was immunolabeled with vimentin and alpha-actinin to determine the cellular origin of Cx43 and was quantified in the following structures: interatrial septum (IAS), His bundle, compact node (CN), lower nodal bundle (LNB), leftward and rightward nodal extensions (LE and RE), and inferior, endocardial, and left-sided transitional cells. Histology revealed two nodal extensions in three of four hearts. Cx43 was found in the myocytes, but not fibroblasts, of the AVJ. LE and CN Cx43 was lower than the IAS (P < 0.05) and the RE, LNB, and His all expressed Cx43 similarly, with approximately half of IAS expression (RE: 44 +/- 36%; LNB: 50 +/- 26%; His: 48 +/- 12%, P = NS compared with IAS). Cx43 levels in transitional cells were similar to the IAS (P = not significant). Cx43 was found in myocytes of the human AVJ, and its expression pattern delineates two separate continuous structures: one consists of the LE and CN with little Cx43, and the other consists of the His, LNB, and RE expressing approximately half the Cx43 of the IAS. The differential Cx43 expression may provide each structure with unique conduction properties, contributing to arrhythmias arising from the AVJ.

Computer Three-dimensional Reconstruction of the Atrioventricular Node

Circulation Research. Apr, 2008  |  Pubmed ID: 18309098

Because of its complexity, the atrioventricular node (AVN), remains 1 of the least understood regions of the heart. The aim of the study was to construct a detailed anatomic model of the AVN and relate it to AVN function. The electric activity of a rabbit AVN preparation was imaged using voltage-dependent dye. The preparation was then fixed and sectioned. Sixty-five sections at 60- to 340-microm intervals were stained for histology and immunolabeled for neurofilament (marker of nodal tissue) and connexin43 (gap junction protein). This revealed multiple structures within and around the AVN, including transitional tissue, inferior nodal extension, penetrating bundle, His bundle, atrial and ventricular muscle, central fibrous body, tendon of Todaro, and valves. A 3D anatomically detailed mathematical model (approximately 13 million element array) of the AVN and surrounding atrium and ventricle, incorporating all cell types, was constructed. Comparison of the model with electric activity recorded in experiments suggests that the inferior nodal extension forms the slow pathway, whereas the transitional tissue forms the fast pathway into the AVN. In addition, it suggests the pacemaker activity of the atrioventricular junction originates in the inferior nodal extension. Computer simulation of the propagation of the action potential through the anatomic model shows how, because of the complex structure of the AVN, reentry (slow-fast and fast-slow) can occur. In summary, a mathematical model of the anatomy of the AVN has been generated that allows AVN conduction to be explored.

Images in Cardiovascular Medicine. Optical Mapping of the Human Atrioventricular Junction

Circulation. Mar, 2008  |  Pubmed ID: 18347223

Effect of Electroporation on Cardiac Electrophysiology

Methods in Molecular Biology (Clifton, N.J.). 2008  |  Pubmed ID: 18370220

Defibrillation shocks are commonly used to terminate life-threatening arrhythmias. According to the excitation theory of defibrillation, such shocks are aimed at depolarizing the membranes of most cardiac cells, resulting in resynchronization of electrical activity in the heart. If shock-induced transmembrane potentials are large enough, they can cause transient tissue damage due to electroporation. In this review, evidence is presented that electroporation of the heart tissue can occur during clinically relevant intensities of the external electrical field and that electroporation can affect the outcome of defibrillation therapy, being both pro- and antiarrhythmic.Here, we present experimental evidence for electroporation in cardiac tissue, which occurs above a threshold of 25 V/cm as evident from propidium iodide uptake, transient diastolic depolarization, and reductions of action potential amplitude and its derivative. These electrophysiological changes can induce tachyarrhythmia, due to conduction block and possibly triggered activity; however, our findings provide the foundation for future design of effective methods to deliver genes and drugs to cardiac tissues, while avoiding possible side effects such as arrhythmia and mechanical stunning.

Spatial Distribution and Extent of Electroporation by Strong Internal Shock in Intact Structurally Normal and Chronically Infarcted Rabbit Hearts

Journal of Cardiovascular Electrophysiology. Oct, 2008  |  Pubmed ID: 18479336

Although life-saving, a strong internal defibrillation shock may temporarily or permanently damage the heart via disruption of cell membranes (electroporation). Spatial extent of electroporation in intact, normal, or infarcted hearts has not been investigated. In this study, shock-induced electroporation in intact rabbit hearts with and without chronic (>4 weeks) left ventricular myocardial infarction (MI) was characterized.

Quantification of Cardiac Fiber Orientation Using Optical Coherence Tomography

Journal of Biomedical Optics. May-Jun, 2008  |  Pubmed ID: 18601522

Heterogeneity in cardiac tissue microstructure is a potential mechanism for the generation and maintenance of arrhythmias. Abnormal changes in fiber orientation increase the likelihood of arrhythmia. We present optical coherence tomography (OCT) as a method to image myofibers in excised intact heart preparations. Three-dimensional (3-D) image sets were gathered from the rabbit right ventricular free wall (RVFW) using a microscope-integrated OCT system. An automated algorithm for fiber orientation quantification in the plane parallel to the wall surface was developed. The algorithm was validated by comparison with manual measurements. Quantifying fiber orientation in the plane parallel to the wall surface from OCT images can be used to help understand the conduction system of the specific sample being imaged.

Quantitative Panoramic Imaging of Epicardial Electrical Activity

Annals of Biomedical Engineering. Oct, 2008  |  Pubmed ID: 18654852

Fluorescent imaging with voltage- and/or calcium-sensitive dyes has revolutionized cardiac physiology research. Here we present improved panoramic imaging for optically mapping electrical activity from the entire epicardium of the Langendorff-perfused rabbit heart. Combined with reconstruction of the 3D heart surface, the functional data can be conveniently visualized on the realistic heart geometry. Methods to quantify the panoramic data set are introduced by first describing a simple approach to mesh the heart in regular grid form. The regular grid mesh provides substrate for easy translation of previously available non-linear dynamics methods for 2D array data. It also simplifies the unwrapping of curved three-dimensional surface to 2D surface for global epicardial visualization of the functional data. The translated quantification methods include activation maps (isochrones), phase maps, phase singularity, and electric stimulus-induced virtual electrode polarization (VEP) maps. We also adapt a method to calculate the conduction velocities on the global epicardial surface by taking the curvature of the heart surface into account.

Bimodal Biophotonic Imaging of the Structure-function Relationship in Cardiac Tissue

Journal of Biomedical Optics. Sep-Oct, 2008  |  Pubmed ID: 19021392

The development of systems physiology is hampered by the limited ability to relate tissue structure and function in intact organs in vivo or in vitro. Here, we show the application of a bimodal biophotonic imaging approach that employs optical coherence tomography and fluorescent imaging to investigate the structure-function relationship at the tissue level in the heart. Reconstruction of cardiac excitation and structure was limited by the depth penetration of bimodal imaging to approximately 2 mm in atrial tissue, and approximately 1 mm in ventricular myocardium. The subcellular resolution of optical coherence tomography clearly demonstrated that microscopic fiber orientation governs the pattern of wave propagation in functionally characterized rabbit sinoatrial and atrioventricular nodal preparations and revealed structural heterogeneities contributing to ventricular arrhythmias. The combination of this bimodal biophotonic imaging approach with histology and/or immunohistochemistry can span multiple scales of resolution for the investigation of the molecular and structural determinants of intact tissue physiology.

Chronaxie of Defibrillation: a Pathway Toward Further Optimization of Defibrillation Waveform?

Journal of Cardiovascular Electrophysiology. Mar, 2009  |  Pubmed ID: 19175836

Resolution of Established Cardiac Hypertrophy and Fibrosis and Prevention of Systolic Dysfunction in a Transgenic Rabbit Model of Human Cardiomyopathy Through Thiol-sensitive Mechanisms

Circulation. Mar, 2009  |  Pubmed ID: 19255346

Cardiac hypertrophy, the clinical hallmark of hypertrophic cardiomyopathy (HCM), is a major determinant of morbidity and mortality not only in HCM but also in a number of cardiovascular diseases. There is no effective therapy for HCM and generally for cardiac hypertrophy. Myocardial oxidative stress and thiol-sensitive signaling molecules are implicated in pathogenesis of hypertrophy and fibrosis. We posit that treatment with N-acetylcysteine, a precursor of glutathione, the largest intracellular thiol pool against oxidative stress, could reverse cardiac hypertrophy and fibrosis in HCM.

Multiple Monophasic Shocks Improve Electrotherapy of Ventricular Tachycardia in a Rabbit Model of Chronic Infarction

Heart Rhythm : the Official Journal of the Heart Rhythm Society. Jul, 2009  |  Pubmed ID: 19560090

We previously showed that the cardioversion threshold (CVT) for ventricular tachycardia (VT) is phase dependent when a single monophasic shock (1MP) is used.

Transient Local Injury Current in Right Ventricular Electrogram After Implantable Cardioverter-defibrillator Shock Predicts Heart Failure Progression

Journal of the American College of Cardiology. Aug, 2009  |  Pubmed ID: 19695461

This study aimed to identify an early marker of functional impairment after an implantable cardioverter-defibrillator (ICD) shock as a predictor of heart failure progression.

Virtual Histology of the Human Heart Using Optical Coherence Tomography

Journal of Biomedical Optics. Sep-Oct, 2009  |  Pubmed ID: 19895104

Optical coherence tomography (OCT) allows for the visualization of micron-scale structures within nontransparent biological tissues. For the first time, we demonstrate the use of OCT in identifying components of the cardiac conduction system and other structures in the explanted human heart. Reconstructions of cardiac structures up to 2 mm below the tissue surface were achieved and validated with Masson Trichrome histology in atrial, ventricular, sinoatrial nodal, and atrioventricular nodal preparations. The high spatial resolution of OCT provides visualization of cardiac fibers within the myocardium, as well as elements of the cardiac conduction system; however, a limiting factor remains its depth penetration, demonstrated to be approximately 2 mm in cardiac tissues. Despite its currently limited imaging depth, the use of OCT to identify the structural determinants of both normal and abnormal function in the intact human heart is critical in its development as a potential aid to intracardiac arrhythmia diagnosis and therapy.

Enhanced Susceptibility to Alternans in a Rabbit Model of Chronic Myocardial Infarction

Conference Proceedings : ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference. 2009  |  Pubmed ID: 19964643

The aim of this study is to examine how structural discontinuity and functional remodeling changes the susceptibility to alternans of action potential duration (APD) in a rabbit model of chronic myocardial infarction (MI). Optical mapping experiments using voltage-sensitive dyes were performed in 14 rabbit hearts. We found that (1) APD alternans starts at a significantly slower pacing rate in hearts with MI (n = 7) than in normal hearts (n = 7), with the original sites of alternans of APD located in the infarct region and infarct adjacent regions. (2) Alternans of activation cycle length (CL) precedes the occurrence of spatially discordant alternans of APD, with the regions of activation CL alternans located in the infarct adjacent regions. Based on these results, we conclude that susceptibility to alternans are significantly enhanced in this rabbit model of chronic MI, and the enhancement is strongly correlated to structural and functional heterogeneity imposed by the infarction.

Membrane Time Constant During Internal Defibrillation Strength Shocks in Intact Heart: Effects of Na+ and Ca2+ Channel Blockers

Journal of Cardiovascular Electrophysiology. Jan, 2009  |  Pubmed ID: 18775052

We assessed defibrillation strength shock-induced changes of the membrane time constant (tau) and membrane potential (DeltaVm) in intact rabbit hearts after administration of lidocaine, a sodium (Na(+)) channel blocker, or nifedipine, a L-type calcium (Ca(2+)) channel blocker.

Structural and Functional Evidence for Discrete Exit Pathways That Connect the Canine Sinoatrial Node and Atria

Circulation Research. Apr, 2009  |  Pubmed ID: 19246679

Surface electrode recordings cannot delineate the activation within the human or canine sinoatrial node (SAN) because they are intramural structures. Thus, the site of origin of excitation and conduction pathway(s) within the SAN of these mammals remains unknown. Canine right atrial preparations (n=7) were optically mapped. The SAN 3D structure and protein expression were mapped using immunohistochemistry. SAN optical action potentials had diastolic depolarization and multiple upstroke components that corresponded to the separate excitations of the node and surface atrial layers. Pacing-induced SAN exit block eliminated atrial optical action potential components but retained SAN optical action potential components. Excitation originated in the SAN (cycle length, 557+/-72 ms) and slowly spread (1.2 to 14 cm/sec) within the SAN, failing to directly excite the crista terminalis and intraatrial septum. After a 49+/-22 ms conduction delay within the SAN, excitation reached the atrial myocardium via superior and/or inferior sinoatrial exit pathways 8.8+/-3.2 mm from the leading pacemaker site. The ellipsoidal 13.7+/-2.8/4.9+/-0.6 mm SAN structure was functionally insulated from the atrium. This insulation coincided with connexin43-negative regions at the borders of the node, connective tissue, and coronary arteries. During normal sinus rhythm, the canine SAN is functionally insulated from the surrounding atrial myocardium except for 2 (or more) narrow superior and inferior sinoatrial exit pathways separated by 12.8+/-4.1 mm. Conduction failure in these sinoatrial exit pathways leads to SAN exit block and is a modulator of heart rate.

Naum Lazarevich Gurvich (1905-1981) and His Contribution to the History of Defibrillation

Cardiology Journal. 2009  |  Pubmed ID: 19387971

Cardioversion: Past, Present, and Future

Circulation. Oct, 2009  |  Pubmed ID: 19841308

Panoramic Imaging Reveals Basic Mechanisms of Induction and Termination of Ventricular Tachycardia in Rabbit Heart with Chronic Infarction: Implications for Low-voltage Cardioversion

Heart Rhythm : the Official Journal of the Heart Rhythm Society. Jan, 2009  |  Pubmed ID: 18996057

Sudden cardiac death due to arrhythmia in the settings of chronic myocardial infarction (MI) is an important clinical problem. Arrhythmic risk post-MI continues indefinitely even if heart failure and acute ischemia are not present due to the anatomic substrate of the scar and border zone (BZ) tissue.

Transmural Dispersion of Repolarization in Failing and Nonfailing Human Ventricle

Circulation Research. Mar, 2010  |  Pubmed ID: 20093630

Transmural dispersion of repolarization has been shown to play a role in the genesis of ventricular tachycardia and fibrillation in different animal models of heart failure (HF). Heterogeneous changes of repolarization within the midmyocardial population of ventricular cells have been considered an important contributor to the HF phenotype. However, there is limited electrophysiological data from the human heart.

Mapping Cardiac Pacemaker Circuits: Methodological Puzzles of the Sinoatrial Node Optical Mapping

Circulation Research. Feb, 2010  |  Pubmed ID: 20133911

Historically, milestones in science are usually associated with methodological breakthroughs. Likewise, the advent of electrocardiography, microelectrode recordings and more recently optical mapping have ushered in new periods of significance of advancement in elucidating basic mechanisms in cardiac electrophysiology. As with any novel technique, however, data interpretation is challenging and should be approached with caution, as it cannot be simply extrapolated from previously used methodologies and with experience and time eventually becomes validated. A good example of this is the use of optical mapping in the sinoatrial node (SAN): when microelectrode and optical recordings are obtained from the same site in myocardium, significantly different results may be noted with respect to signal morphology and as a result have to be interpreted by a different set of principles. Given the rapid spread of the use of optical mapping, careful evaluation must be made in terms of methodology with respect to interpretation of data gathered by optical sensors from fluorescent potential-sensitive dyes. Different interpretations of experimental data may lead to different mechanistic conclusions. This review attempts to address the origin and interpretation of the "double component" morphology in the optical action potentials obtained from the SAN region. One view is that these 2 components represent distinctive signals from the SAN and atrial cells and can be fully separated with signal processing. A second view is that the first component preceding the phase 0 activation represents the membrane currents and intracellular calcium transients induced diastolic depolarization from the SAN. Although the consensus from both groups is that ionic mechanisms, namely the joint action of the membrane and calcium automaticity, are important in the SAN function, it is unresolved whether the double-component originates from the recording methodology or represents the underlying physiology. This overview aims to advance a common understanding of the basic principles of optical mapping in complex 3D anatomic structures.

Anatomy and Electrophysiology of the Human AV Node

Pacing and Clinical Electrophysiology : PACE. Jun, 2010  |  Pubmed ID: 20180918

The atrioventricular node (AVN) has mystified generations of investigators over the last century and continues today to be at the epicenter of debates among anatomists, experimentalists, and electrophysiologists. Over the years, discrepancies have remained in regard to correlating components of AVN structure to function, as evidenced by studies from microelectrodes, optical mapping, and the electrophysiology laboratory. Historically, the AVN has been defined by classical histological methods; however, with recent advances in molecular biology techniques, a more precise characterization of structure is becoming attainable. Distinct molecular compartments are becoming apparent based on connexin staining and genotyping, providing new insight into previously characterized functional aspects of the AVN and its surrounding structures. Advances in optical mapping have provided a unique opportunity for correlating structure and function--unmasking properties of the native AVN pacemaker and providing further insight into basic mechanisms involved in AV conduction. Additionally, procurement of explanted human hearts have provided a unique opportunity to further characterize the human AVN structurally and functionally with both molecular biology techniques and optical mapping. With the elucidation of basic elements of both structure and function via molecular investigation and optical mapping, new opportunities are becoming apparent in utilizing the unique properties of the AVN for pursuing novel clinical applications relevant to clinical electrophysiology.

Functional Anatomy of the Murine Sinus Node: High-resolution Optical Mapping of Ankyrin-B Heterozygous Mice

American Journal of Physiology. Heart and Circulatory Physiology. Aug, 2010  |  Pubmed ID: 20525877

The mouse is widely used as a genetic platform to investigate the molecular mechanisms of sinoatrial node (SAN) pacemaking. Recently, it has been shown that isolated SAN cells from the ankyrin-B (AnkB)-deficient mice display severe pacemaking dysfunction similar to individuals harboring ankyrin 2 allele variants. However, these results have been limited to isolated SAN cells only and thus did not evaluate the functional anatomy of the widely distributed atrial pacemaker complex (e.g., the dynamic interaction of primary and subsidiary pacemakers). We studied pacemaker function in an intact mouse atrial preparation, which included the SAN, atrioventricular junction (AVJ), and both atria, excluding most of the septum. Optical mapping with a voltage-sensitive dye and CMOS camera ULTIMA-L was used to map spontaneous pacemaker activity with or without autonomic modulation in wild-type (WT) mice (n = 7) and in the AnkB heterozygous (AnkB(+/-); n = 9) mouse model of human SAN disease. In WT mice, isoproterenol accelerated the SAN rate (for 10 microM: from 325 + or - 19 to 510 + or - 33 beat/min, P < 0.01) and shifted the leading pacemaker site superiorly by 0.77 + or - 0.11 mm within the SAN. ACh decreased the SAN rate (from 333 + or - 26 to 96 + or - 22 beats/min, P < 0.01) and shifted the leading pacemaker either inferiorly within the SAN or abruptly toward the AVJ. After isoproterenol, AnkB(+/-) mice exhibited a larger beat-to-beat variability (SD of a cycle length: 13.4 + or - 3.6 vs. 2.5 + or - 0.8 ms, P < 0.01 vs. WT mice), disorganized shift of the leading pacemaker (2.04 + or - 0.37 mm, P < 0.05 vs. WT mice), and competing multiple pacemakers, resulting in beat-to-beat changes of the leading pacemaker location site between the SAN and AVJ regions. Notably, AnkB(+/-) mice also displayed a reduced sensitivity to ACh (rate slowing by 32 + or - 12% vs. 67 + or - 4%, P < 0.05, AnkB(+/-) vs. WT mice, respectively). In conclusion, AnkB dysfunction results in SAN abnormalities in an isolated mouse atria preparation. While AnkB dysfunction dramatically alters single SAN cell function, the mechanisms underlying cardiac automaticity are clearly complex, and phenotypes may be partially compensated by the dynamic interaction of cells within the pacemaker complex. These new findings highlight the importance of the functional anatomy of the entire atrial distributed pacemaker complex, including the SAN and AVJ, and clearly demonstrate the role of AnkB in cardiac automaticity.

Mechanisms of Fibrillation: Neurogenic or Myogenic? Reentrant or Focal? Multiple or Single? Still Puzzling After 160 Years of Inquiry

Journal of Cardiovascular Electrophysiology. Nov, 2010  |  Pubmed ID: 20550608

Complex Interactions Between the Sinoatrial Node and Atrium During Reentrant Arrhythmias in the Canine Heart

Circulation. Aug, 2010  |  Pubmed ID: 20697021

Numerous studies implicate the sinoatrial node (SAN) as a participant in atrial arrhythmias, including atrial flutter (AFL) and atrial fibrillation (AF). However, the direct role of the SAN has never been described.

Optical Mapping of the Isolated Coronary-perfused Human Sinus Node

Journal of the American College of Cardiology. Oct, 2010  |  Pubmed ID: 20946995

We sought to confirm our hypothesis that the human sinoatrial node (SAN) is functionally insulated from the surrounding atrial myocardium except for several exit pathways that electrically bridge the nodal tissue and atrial myocardium.

Molecular Remodeling of Ion Channels, Exchangers and Pumps in Atrial and Ventricular Myocytes in Ischemic Cardiomyopathy

Channels (Austin, Tex.). Mar-Apr, 2010  |  Pubmed ID: 20090424

Existing molecular knowledge base of cardiovascular diseases is rudimentary because of lack of specific attribution to cell type and function. The aim of this study was to investigate cell-specific molecular remodeling in human atrial and ventricular myocytes associated with ischemic cardiomyopathy. Our strategy combines two technological innovations, laser-capture microdissection of identified cardiac cells in selected anatomical regions of the heart and splice microarray of a narrow catalog of the functionally most important genes regulating ion homeostasis. We focused on expression of a principal family of genes coding for ion channels, exchangers and pumps (CE&P genes) that are involved in electrical, mechanical and signaling functions of the heart and constitute the most utilized drug targets. We found that (1) CE&P genes remodel in a cell-specific manner: ischemic cardiomyopathy affected 63 CE&P genes in ventricular myocytes and 12 essentially different genes in atrial myocytes. (2) Only few of the identified CE&P genes were previously linked to human cardiac disfunctions. (3) The ischemia-affected CE&P genes include nuclear chloride channels, adrenoceptors, cyclic nucleotide-gated channels, auxiliary subunits of Na(+), K(+) and Ca(2+) channels, and cell-surface CE&Ps. (4) In both atrial and ventricular myocytes ischemic cardiomyopathy reduced expression of CACNG7 and induced overexpression of FXYD1, the gene crucial for Na(+) and K(+) homeostasis. Thus, our cell-specific molecular profiling defined new landmarks for correct molecular modeling of ischemic cardiomyopathy and development of underlying targeted therapies.

Differential K(ATP) Channel Pharmacology in Intact Mouse Heart

Journal of Molecular and Cellular Cardiology. Jan, 2010  |  Pubmed ID: 19744493

Classically, cardiac sarcolemmal K(ATP) channels have been thought to be composed of Kir6.2 (KCNJ11) and SUR2A (ABCC9) subunits. However, the evidence is strong that SUR1 (sulfonylurea receptor type 1, ABCC8) subunits are also expressed in the heart and that they play a significant functional role in the atria. To examine this further, we have assessed the effects of isotype-specific potassium channel-opening drugs, diazoxide (specific to SUR1>SUR2A) and pinacidil (SUR2A>SUR1), in intact hearts from wild-type mice (WT, n=6), SUR1(-/-) (n=6), and Kir6.2(-/-) mice (n=5). Action potential durations (APDs) in both atria and ventricles were estimated by optical mapping of the posterior surface of Langendorff-perfused hearts. To confirm the atrial effect of both openers, isolated atrial preparations were mapped in both WT (n=4) and SUR1(-/-) (n=3) mice. The glass microelectrode technique was also used to validate optical action potentials. In WT hearts, diazoxide (300 microM) decreased APD in atria (from 33.8+/-1.9 ms to 24.2+/-1.1 ms, p<0.001) but was without effect in ventricles (APD 60.0+/-7.6 ms vs. 60.8+/-7.5 ms, respectively, NS), consistent with an atrial-specific role for SUR1. The absence of SUR1 resulted in loss of efficacy of diazoxide in SUR1(-/-) atria (APD 36.8+/-1.9 ms vs. 36.8+/-2.8 ms, respectively, NS). In contrast, pinacidil (300 microM) significantly decreased ventricular APD in both WT and SUR1(-/-) hearts (from 60.0+/-7.6 ms to 29.8+/-3.5 ms in WT, p<0.001, and from 63.5+/-2.1 ms to 24.8+/-3.8 ms in SUR1(-/-), p<0.001), but did not decrease atrial APD in either WT or SUR1(-/-) hearts. Glibenclamide (10 microM) reversed the effect of pinacidil in ventricles and restored APD to control values. The absence of Kir6.2 subunits in Kir6.2(-/-) hearts resulted in loss of efficacy of both openers (APD 47.2+/-2.2 ms vs. 47.6+/-2.1 ms and 50.8+/-2.4 ms, and 90.6+/-5.7 ms vs. 93.2+/-6.5 ms and 117.3+/-6.4 ms, for atria and ventricle in control versus diazoxide and pinacidil, respectively). Collectively, these results indicate that in the same mouse heart, significant differential K(ATP) pharmacology in atria and ventricles, resulting from SUR1 predominance in forming the atrial channel, leads to differential effects of potassium channel openers on APD in the two chambers.

Structured Light Imaging of Epicardial Mechanics

Conference Proceedings : ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference. 2010  |  Pubmed ID: 21095816

There is a need for accurate measurements of mechanical strain and motion of the heart both in vitro and in vivo. We have developed a new structured-light imaging system capable of epicardial shape measurement at 333 fps at a resolution of 768 × 768 pixels. Here we present proof-of-concept data from our system applied to a beating rabbit heart in vitro to measure epicardial mechanics. This method will allow high resolution mapping of epicardial strain and virtual immobilization of the heart for removal of motion artifacts from epicardial recordings with fluorescence dyes. This will allow mapping of transmembrane potential and calcium transients in a beating heart, including in vivo.

Multiscale Imaging of the Human Heart: Building the Foundation for Human Systems Physiology and Translational Medicine

Conference Proceedings : ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference. 2010  |  Pubmed ID: 21095821

The development of human cardiovascular systems physiology is inhibited by the lack of multiscale functional physiological data, which represents human heart physiology at the molecular, cellular, tissue, organ, and system levels. We have developed an experimental approach to study explanted human hearts in vitro at multiple physiological scales with a wide array of imaging modalities. This approach has already yielded data indicating significant differences between animal models of diseases and actual human heart disease. Our data provides a quantitative foundation for multiscale physiological models of the cardiovascular system and will allow improvement in translation of medical technology and pharmacology from animal models to therapy.

Transmural Heterogeneity and Remodeling of Ventricular Excitation-contraction Coupling in Human Heart Failure

Circulation. May, 2011  |  Pubmed ID: 21502574

Excitation-contraction (EC) coupling is altered in end-stage heart failure. However, spatial heterogeneity of this remodeling has not been established at the tissue level in failing human heart. The objective of this article was to study functional remodeling of excitation-contraction coupling and calcium handling in failing and nonfailing human hearts.

Effects of KATP Channel Openers Diazoxide and Pinacidil in Coronary-perfused Atria and Ventricles from Failing and Non-failing Human Hearts

Journal of Molecular and Cellular Cardiology. Aug, 2011  |  Pubmed ID: 21586291

This study compared the effects of ATP-regulated potassium channel (K(ATP)) openers, diazoxide and pinacidil, on diseased and normal human atria and ventricles. We optically mapped the endocardium of coronary-perfused right (n=11) or left (n=2) posterior atrial-ventricular free wall preparations from human hearts with congestive heart failure (CHF, n=8) and non-failing human hearts without (NF, n=3) or with (INF, n=2) infarction. We also analyzed the mRNA expression of the K(ATP) targets K(ir)6.1, K(ir)6.2, SUR1, and SUR2 in the left atria and ventricles of NF (n=8) and CHF (n=4) hearts. In both CHF and INF hearts, diazoxide significantly decreased action potential durations (APDs) in atria (by -21±3% and -27±13%, p<0.01) and ventricles (by -28±7% and -28±4%, p<0.01). Diazoxide did not change APD (0±5%) in NF atria. Pinacidil significantly decreased APDs in both atria (-46 to -80%, p<0.01) and ventricles (-65 to -93%, p<0.01) in all hearts studied. The effect of pinacidil on APD was significantly higher than that of diazoxide in both atria and ventricles of all groups (p<0.05). During pinacidil perfusion, burst pacing induced flutter/fibrillation in all atrial and ventricular preparations with dominant frequencies of 14.4±6.1 Hz and 17.5±5.1 Hz, respectively. Glibenclamide (10 μM) terminated these arrhythmias and restored APDs to control values. Relative mRNA expression levels of K(ATP) targets were correlated to functional observations. Remodeling in response to CHF and/or previous infarct potentiated diazoxide-induced APD shortening. The activation of atrial and ventricular K(ATP) channels enhances arrhythmogenicity, suggesting that such activation may contribute to reentrant arrhythmias in ischemic hearts.

Anatomic Localization and Autonomic Modulation of Atrioventricular Junctional Rhythm in Failing Human Hearts

Circulation. Arrhythmia and Electrophysiology. Aug, 2011  |  Pubmed ID: 21646375

The structure-function relationship in the atrioventricular junction (AVJ) of various animal species has been investigated in detail; however, less is known about the human AVJ. In this study, we performed high-resolution optical mapping of the human AVJ (n = 6) to define its pacemaker properties and response to autonomic stimulation.

Role of Pyk2 in Cardiac Arrhythmogenesis

American Journal of Physiology. Heart and Circulatory Physiology. Sep, 2011  |  Pubmed ID: 21666110

Proline-rich tyrosine kinase 2 (Pyk2) is a nonreceptor protein kinase regulated by intracellular Ca(2+), CaMK, and PKC and can be activated by different stress signals involved in heart failure. However, Pyk2 has not been investigated in the human heart, and the functional role of Pyk2 signaling at the whole heart level has not been elucidated. We hypothesize that Ca(2+)-dependent activation of Pyk2 is involved in cardiac electrophysiology. We examined the expression of Pyk2 in nonfailing versus ischemic and nonischemic failing human hearts (n = 6 hearts/group). To investigate Pyk2 function, we optically mapped perfused hearts from wild-type (WT; n = 7) and knockout (Pyk2(-/-); n = 8) mice during autonomic stimulation. Experiments were done in control mice and after 1 wk of transverse aortic constriction. We used the Illumina beadarray approach for transcriptional profiling of WT and Pyk2(-/-) mouse ventricles. Western blot analysis revealed a doubling of Pyk2 activation in nonischemic failing versus nonfailing human hearts. In mouse hearts, we observed a much higher probability of ventricular tachyarrhythmia during ACh perfusion in Pyk2(-/-) versus WT mice. Parasympathetic stimulation resulted in a dose-dependent decrease of atrial action potential duration (APD) in both WT and Pyk2(-/-) mice, whereas in ventricles it induced APD shortening in Pyk2(-/-) mice but not in WT mice. Deficiency of Pyk2 abolished ACh-induced prolongation of atrioventricular delay in Pyk2(-/-) mouse hearts but did not affect heart rate. Lower mRNA and protein levels of sarco(endo)plasmic reticulum Ca(2+)-ATPase 2 and higher mRNA levels of Na(+)/Ca(2+) exchanger 1 were detected in Pyk2(-/-) hearts compared with WT hearts. The transverse aortic constriction protocol did not change the phenotype. In conclusion, our results indicate a protective role of Pyk2 with respect to ventricular tachyarrhythmia during parasympathetic stimulation by regulation of gene expression related to Ca(2+) handling. We hypothesize that activation of Pyk2 in the human heart during heart failure may contribute to protection against arrhythmia.

Spatiotemporal Control of Heart Rate in a Rabbit Heart

Journal of Electrocardiology. Nov-Dec, 2011  |  Pubmed ID: 21937057

Sinoatrial node is responsible for the origin of the wave of excitation, which spreads throughout the heart and orchestrates cardiac contraction via calcium-mediated excitation-contraction coupling. P wave represents the spread of excitation in the atria. It is well known that the autonomic nervous system controls the heart rate by dynamically altering both cellular ionic fluxes and the anatomical location of the leading pacemaker. In this study, we used isolated rabbit right atria and mathematical model of the pacemaker region of the rabbit heart. Application of isoproterenol resulted in dose-dependent acceleration of the heart rate and superior shift of the leading pacemaker. In the mathematical model, such behavior could be reproduced by a gradient of expression in β1-adrenergic receptors along the superior-inferior axis. Application of acetylcholine resulted in preferentially inferior shift of pacemaker and slowing of the heart rate. The mathematical model reproduced this behavior with imposing a gradient of expression of acetylcholine-sensitive potassium channel. We conclude that anatomical shift of the leading pacemaker in the rabbit heart could be achieved through gradient of expression of β1-adrenergic receptors and I(K,ACh).

Low-energy Multistage Atrial Defibrillation Therapy Terminates Atrial Fibrillation with Less Energy Than a Single Shock

Circulation. Arrhythmia and Electrophysiology. Dec, 2011  |  Pubmed ID: 21980076

Implantable device therapy of atrial fibrillation (AF) is limited by pain from high-energy shocks. We developed a low-energy multistage defibrillation therapy and tested it in a canine model of AF.

Termination of Sustained Atrial Flutter and Fibrillation Using Low-voltage Multiple-shock Therapy

Heart Rhythm : the Official Journal of the Heart Rhythm Society. Jan, 2011  |  Pubmed ID: 20969974

Defibrillation therapy for atrial fibrillation (AF) and flutter (AFl) is limited by pain induced by high-energy shocks. Thus, lowering the defibrillation energy for AFl/AF is desirable.

The Role of Dynamic Instability and Wavelength in Arrhythmia Maintenance As Revealed by Panoramic Imaging with Blebbistatin Vs. 2,3-butanedione Monoxime

American Journal of Physiology. Heart and Circulatory Physiology. Jan, 2012  |  Pubmed ID: 22037192

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.

Longitudinal Study of Cardiac Remodelling in Rabbits Following Infarction

The Canadian Journal of Cardiology. Jan, 2012  |  Pubmed ID: 22265993

BACKGROUND: Cardiac remodelling following myocardial infarction (MI) is a complex, dynamic process. There have been few longitudinal studies of these changes. METHODS: A 2-dimensional transthoracic echocardiography was performed on 20 rabbits, before and 1, 2, 4, 8, and 12 weeks after MI (n = 14) and twice for controls (n = 6). Chronic left ventricular (LV) infarct size was histologically characterized and correlated with mechanical function. A linear mixed model was used to analyze longitudinal and infarct size-related changes in LV end-systolic volume (ESV), end-diastolic volume (EDV), ejection fraction (EF), sphericity, circumferential strain, and wall motion score index. RESULTS: Mean LV infarct size was 28.9% ± 9.3%. After MI, rapid remodelling occurred in LVESV, LVEF, and sphericity for 2 weeks and LVEDV for 4 weeks, with slower changes afterwards. LV infarct size correlated with LVESV (r = 0.76), LVEDV (r = 0.71), and LVEF (r = 0.69). Larger infarcts resulted in greater LVESV dilation (P = 0.04) and faster LVEDV (P < 0.01), LVEF (P < 0.01), and sphericity (P < 0.01) remodelling. Apical global circumferential strain and wall motion score index increased for 1 week, then stabilized, regardless of infarct size, and apical global circumferential strain was correlated with apical infarction (r = 0.58). Additionally, regional circumferential strain decreased in segments with severe (> 80%) infarction more quickly (P < 0.01) and by a greater degree (P = 0.04) compared with segments with minor (< 20%) infarction. CONCLUSIONS: The most dynamic remodelling of cardiac function in this model occurred during the first 4 weeks, stabilizing thereafter, with changes maintained up to 12 weeks. Infarct size affected both the early rate and long-term extent of mechanical remodelling.

Three Potential Mechanisms for Failure of HIFU Ablation in Cardiac Tissue

Circulation. Arrhythmia and Electrophysiology. Feb, 2012  |  Pubmed ID: 22322367

BACKGROUND: -High Intensity Focused Ultrasound (HIFU) has been introduced for treatment of cardiac arrhythmias, because it offers the ability to create rapid tissue modification in confined volumes without directly contacting the myocardium. In spite of the benefits of HIFU, a number of limitations have been reported, which hindered its clinical adoption. METHODS AND RESULTS: -In this study, we used a multimodal approach to evaluate thermal and non-thermal effects of HIFU in cardiac ablation. We designed a computer-controlled system capable of simultaneous fluorescence mapping and HIFU ablation. Using this system, linear lesions were created in isolated rabbit atria (n = 6) and point lesions were created in the ventricles of whole-heart (n = 6) preparations by applying HIFU at clinical doses (4-16W). Additionally, we evaluate the gap size in ablation lines necessary for conduction in atrial preparations (n = 4). The voltage sensitive dye di-4-ANEPPS was used to assess functional damage produced by HIFU. Optical coherence tomography and general histology were used to evaluate lesion extent. Conduction block was achieved in 1 (17%) of 6 atrial preparations with a single ablation line. Following 10 minutes of rest, 0 (0%) of 6 atrial preparations demonstrated sustained conduction block from a single ablation line. Tissue displacement of 1-3mm was observed during HIFU application due to acoustic radiation force along the lesion line. Additionally, excessive acoustic pressure and high temperature from HIFU generated cavitation causing macroscopic tissue damage. A minimum gap size of 1.5mm was found to conduct electrical activity. CONCLUSIONS: -This study identified three potential mechanisms responsible for the failure of HIFU ablation in cardiac tissues. Both acoustic radiation force and acoustic cavitation in conjunction with inconsistent thermal deposition can increase the risk of lesion discontinuity and result in gap sizes that promote ablation failure.

Long-term Culture of HL-1 Cardiomyocytes in Modular Poly(ethylene Glycol) Microsphere-based Scaffolds Crosslinked in the Phase-separated State

Acta Biomaterialia. Jan, 2012  |  Pubmed ID: 21920469

Poly(ethylene glycol) (PEG) microspheres were assembled around HL-1 cardiomyocytes to produce highly porous modular scaffolds. In this study we took advantage of the immiscibility of PEG and dextran to improve upon our previously described modular scaffold fabrication methods. Phase separating the PEG microspheres in dextran solutions caused them to rapidly deswell and crosslink together, eliminating the need for serum protein-based crosslinking. This also led to a dramatic increase in the stiffness of the scaffolds and greatly improved the handling characteristics. HL-1 cardiomyocytes were present during microsphere crosslinking in the cytocompatible dextran solution, exhibiting high cell viability following scaffold formation. Over the course of 2 weeks a 9-fold expansion in cell number was observed. The cardiac functional markers sarcomeric α-actinin and connexin 43 were expressed at 13 and 24 days after scaffold formation. HL-1 cells were spontaneously depolarizing 38 days after scaffold formation, which was visualized by confocal microscopy using a calcium-sensitive dye. Electrical stimulation resulted in synchronization of activation peaks throughout the scaffolds. These findings demonstrate that PEG microsphere scaffolds fabricated in the presence of dextran can support the long-term three-dimensional culture of cells, suggesting applications in cardiovascular tissue engineering.