Katherine M. Kavanagh

Complete heart block and sudden death in mice overexpressing calreticulin

The expression of calreticulin, a Ca(2+)-binding chaperone of the endoplasmic reticulum, is elevated in the embryonic heart, and because of impaired cardiac development, knockout of the Calreticulin gene is lethal during embryogenesis. The elevated expression is downregulated after birth. Here we have investigated the physiological consequences of continued high expression of calreticulin in the postnatal heart, by producing transgenic mice that overexpress the protein in the heart. These transgenic animals exhibit decreased systolic function and inward I(Ca,L), low levels of connexin43 and connexin40, sinus bradycardia, and prolonged atrioventricular (AV) node conduction followed by complete heart block and sudden death. We conclude that postnatal downregulation of calreticulin is essential in the development of the cardiac conductive system, in particular in the sinus and AV nodes, when an inward Ca(2+) current is required for activation. This work identifies a novel pathway of events, leading to complete heart block and sudden cardiac death, which involves high expression of calreticulin in the heart.

In vivo temporal and spatial distribution of depolarization and repolarization and the illusive murine T wave

This study assessed in vivo temporal and spatial electrophysiological properties of murine hearts and the effect of manipulation of transmural action potential durations (APDs) on T wave morphology. Monophasic action potentials (MAPs) were acquired from multiple left ventricular sites. All MAPs exhibited a plateau phase, with a spike and dome appearance being present in epicardial recordings. Activation occurred from endocardial apex to epicardial apex and apex to base while repolarization occurred from base (shortest 90 η0 level of repolarization (MAP90), 95.4 ± 8.9 ms) to apex and epicardium to endocardium (longest MAP90, 110.77 ± 10.6 ms). The peak of phase 0 of the epicardial base MAP correlated with the return to baseline of the initial and usually dominant waveform of the QRS and the onset of the second usually smaller wave, which clearly occurred in early repolarization, thus establishing where depolarization ended and repolarization began on the murine ECG. This second waveform was similar to the J wave seen in larger animals. Despite temporal and spatial electrophysiological similarities, a T wave is frequently not seen on a murine ECG. There are several determinants of T wave morphology, including transmural activation time, slope of phase 3 repolarization and differences in epicardial, endocardial and M cell APDs. Experimental manipulation of murine transmural gradients by shortening epicardial MAP90 to 84% of endocardial MAP90 the epicardial/endocardial ratio in larger mammals when a positive T wave is present, resulted in a positive murine T wave. Thus, manipulation of the transmural gradients such that they are similar to larger mammals can result in T waves with similar morphology.

Autoimmune cardiomyopathy and heart block develop spontaneously in HLA-DQ8 transgenic IAβ knockout NOD mice

A line of nonobese diabetic (NOD) mice expressing the human diabetes-associated HLA-DQ8 transgene in the absence of mouse IAβ failed to show spontaneous insulitis or diabetes, but rather developed dilated cardiomyopathy, leading to early death from heart failure. Pathology in these animals results from an organ- and cell-specific autoimmune response against normal cardiomyoctes in the atrial and ventricular walls, as well as against very similar myocytes present in the outermost muscle layer surrounding the pulmonary veins. Progression of the autoimmune process could be followed by serial ECG measurements; irradiation of young animals significantly delayed disease progression, and this effect could be reversed by adoptive transfer of splenocytes taken from older animals with complete heart block. Disease progression could also be blocked by cyclosporin A treatment, but was accelerated by injection of complete Fruends adjuvant. The constellation of findings of spontaneously arising destructive focal lymphocytic infiltrates within the myocardium, rising titers of circulating anticardiac autoantibodies, dilation of the cardiac chambers, and gradual progression to end-stage heart failure bears a striking resemblance to what is seen in humans with idiopathic dilated cardiomyopathy, a serious and often life-threatening medical condition. This transgenic strain provides a highly relevant animal model for human autoimmune myocarditis and postinflammatory dilated cardiomyopathy.

Significance of inwardly directed transmembrane current in determination of local myocardial electrical activation during ventricular fibrillation

Ventricular fibrillation (VF) is the principle cardiac rhythm disorder responsible for sudden cardiac death in humans. The accurate determination of local cardiac activation during VF is essential for its mechanistic elucidation. This has been hampered by the rapidly changing and markedly heterogeneous electrophysiological nature of VF. These difficulties are manifested when attempting to differentiate true propagating electrical activity from electrotonic signals and when identifying local activation from complex and possibly fractionated electrograms. The purpose of this investigation was to test the hypothesis that the presence of a balanced inwardly and outwardly directed transmembrane charge, obtained from the ratio of the inward to outward area under the cardiac transmembrane current curve (-/+ Im area), could reliably differentiate propagating from electrotonic deflections during VF. To test this hypothesis, we applied a recently described technique for the in vivo estimation of the transmembrane current (Im) during cardiac activation. A 17-element orthogonal epicardial electrode array was combined with an immediately adjacent optical fiber array to record electrical and optically coupled transmembrane potential signals during VF. Recordings were obtained during electrically induced VF in six dogs to determine the Im associated with activation and the time course of repolarization, as well as unipolar electrograms and bipolar electrograms recorded at multiple center-to-center interelectrode distances from 0.2 to 3 mm. Propagating local activations were associated with the presence of an easily identified inwardly directed Im, with a balanced inward and outward charge (-/+ Im area approximately 1.0). Electrotonic wave-forms lacked this inward Im (-/+ Im area approximately 0.0). Normal Na(+)-mediated inward currents were directly demonstrated to be responsible for some activations during VF.

Electrophysiologic properties and ventricular fibrillation in normal and myopathic hearts

This study tests the hypothesis that moderate myocardial dysfunction is associated with altered myocardial anisotropic properties and structurally altered ventricular fibrillation (VF). Mongrel dogs were randomized to either a control group or a group that was rapidly paced at 250 beats/min until the left ventricular ejection fraction was < or = 40%. Changes in anisotropic properties and the electrical characteristics of VF associated with the development of moderate myocardial dysfunction were assessed by microminiature epicardial mapping studies. In vivo conduction, refractory periods, and repolarization times were prolonged in both longitudinal and transverse directions in myopathic animals versus controls. VF was different in myopathic versus control animals. There were significantly more conducted deflections during VF in normal hearts compared with myopathic hearts. Propagated deflection-to-deflection intervals during VF were significantly longer in myopathic hearts compared with controls (125.5 +/- 49.06 versus 103.4 +/- 32.9 ms, p = 0.009). There were no abnormalities in cell size, cell shape, or the number of intercellular gap junctions and there was no detectable change in the expression of the gap junction proteins Cx43 and Cx45. Moderate myocardial dysfunction is associated with significant electrophysiological abnormalities in the absence of changes in myocardial cell morphology or intercellular connections, suggesting a functional abnormality in cell-to-cell communication.

Epicardial cardiac source-field behavior

The accurate determination of the spatial distribution of cardiac electrophysiological state is essential for the mechanistic assessment of cardiac arrhythmias in both clinical and experimental cardiac electrophysiological laboratories. The authors describe three fundamental cardiac source-field relationships: 1) activation fields; 2) electrotonic fields; and 3) volume conductor fields. The three cases are described analytically and illustrated with experimentally obtained canine cardiac recordings that capitalize on a recently formulated technique for in vivo cardiac transmembrane current estimation.<<ETX>>

Development of a nonlinearly deterministic signal generator for real time chaos control testing

Chaotic dynamics have been successfully exploited in feedback control strategies. Recently, the authors have described the existence of unstable fixed points during ventricular fibrillation (VF), similar to those previously used in chaos control. Prior to embarking on control attempts of VF, a signal source was designed with known nonlinearly deterministic dynamics with well characterized stable and unstable manifolds as well as fixed point location. This ensured that the complex real-time data acquisition/analysis necessary to attempt such control was feasible for the interval durations typical of VF. A signal source was designed that can operate on any IBM compatible PC via the parallel port, and generates a waveform whose inter-event interval is nonlinearly deterministic with timing resolution of 0.1 /spl mu/sec with a range from 20 msec to 1.5 sec, independent of PC clock speed.

Estimate of electromotive surface dimension during ventricular fibrillation

The accurate identification of local activation from extracellular recording electrodes attached to the beating heart is critical for the development of a mechanistic understanding of serious cardiac rhythm disturbances in both experimental animals and man. One of the most complex of these rhythm disturbances is ventricular fibrillation, a common cause of sudden cardiac death in man. When using bipolar extracellular recording electrodes to detect local activation, the spatial extent of the upstroke of the action potentials that generate the extracellular waveform determines the optimal interelectrode distances for the bipolar electrodes. This spatial length has been well characterized for normal cardiac conduction at around 1.0 mm on the epicardial surface. We found that for many of the activations occurring during early ventricular fibrillation this length is similarly around 1.0 mm.

Pulmonary edema postcardioversion: A potential calcium signalling problem

The present report describes an unusual case of pulmonary edema after adenosine cardioversion of a supraventricular tachycardia. Despite a structurally normal heart, a 52-year-old woman presented with pulmonary edema on two separate occasions, having had her atrioventricular nodal re-entrant tachycardia terminated with 12 mg of intravenous adenosine. A third similar episode of tachycardia that was terminated with verapamil was not complicated by pulmonary edema.

Nonlinear Synchronization Analysis of Spatiotemporal Heart Data

A high‐speed video camera and voltage‐sensitive dyes were used to acquire high resolution (80×80 pixels) and high‐speed (500 μs/frame) optical signals of ventricular fibrillation in a Langendorff‐perfused porcine heart. The resulting spatiotemporal dynamics were recorded before and after the application of a defibrillation shock in order to study the mechanism of defibrillation failure. We calculate nonlinear synchronization index measures to qualify the evolution of different types of activity on the heart surface (focal, reentry). We observe changes with time in the spatial distribution of the first Fourier mode, showing that two main types of activity compete on the heart surface during a failed defibrillation.