A fundamental evaluation of the electrical properties and function of cardiac transverse tubules.

This work discusses active and passive electrical properties of transverse (T-)tubules in ventricular cardiomyocytes to understand the physiological roles of T-tubules. T-tubules are invaginations of the lateral membrane that provide a large surface for calcium-handling proteins to facilitate sarcomere shortening. Higher heart rates correlate with higher T-tubular densities in mammalian ventricular cardiomyocytes. We assess ion dynamics in T-tubules and the effects of sodium current in T-tubules on the extracellular potential, which leads to a partial reduction of the sodium current in deep segments of a T-tubule. We moreover reflect on the impact of T-tubules on macroscopic conduction velocity, integrating fundamental principles of action potential propagation and conduction. We also theoretically assess how the conduction velocity is affected by different T-tubular sodium current densities. Lastly, we critically assess literature on ion channel expression to determine whether action potentials can be initiated in T-tubules.

Uniaxial strain of cultured mouse and rat cardiomyocyte strands slows conduction more when its axis is parallel to impulse propagation than when it is perpendicular.

AIM: Cardiac tissue deformation can modify tissue resistance, membrane capacitance and ion currents and hence cause arrhythmogenic slow conduction. Our aim was to investigate whether uniaxial strain causes different changes in conduction velocity (θ) when the principal strain axis is parallel vs perpendicular to impulse propagation. METHODS: Cardiomyocyte strands were cultured on stretchable custom microelectrode arrays, and θ was determined during steady-state pacing. Uniaxial strain (5%) with principal axis parallel (orthodromic) or perpendicular (paradromic) to propagation was applied for 1 minute and controlled by imaging a grid of markers. The results were analysed in terms of cable theory. RESULTS: Both types of strain induced immediate changes of θ upon application and release. In material coordinates, orthodromic strain decreased θ significantly more (P < .001) than paradromic strain (2.2 ± 0.5% vs 1.0 ± 0.2% in n = 8 mouse cardiomyocyte cultures, 2.3 ± 0.4% vs 0.9 ± 0.5% in n = 4 rat cardiomyocyte cultures, respectively). The larger effect of orthodromic strain can be explained by the increase in axial myoplasmic resistance, which is not altered by paradromic strain. Thus, changes in tissue resistance substantially contributed to the changes of θ during strain, in addition to other influences (eg stretch-activated channels). Besides these immediate effects, the application of strain also consistently initiated a slow progressive decrease in θ and a slow recovery of θ upon release. CONCLUSION: Changes in cardiac conduction velocity caused by acute stretch do not only depend on the magnitude of strain but also on its orientation relative to impulse propagation. This dependence is due to different effects on tissue resistance.

Activation of electrical triggers of atrial fibrillation in hyperthyroidism.

CONTEXT: A shortening of the atrial refractory period has been considered as the main mechanism for the increased risk of atrial fibrillation in hyperthyroidism. However, other important factors may be involved. OBJECTIVE: Our objective was to determine the activity of abnormal supraventricular electrical depolarizations in response to elevated thyroid hormones in patients without structural heart disease. PATIENTS AND DESIGN: Twenty-eight patients (25 females, three males, mean age 43+/-11 yr) with newly diagnosed and untreated hyperthyroidism were enrolled in a prospective trial after exclusion of heart disease. Patients were followed up for 16 +/- 6 months and studied at baseline and 6 months after normalization of serum TSH levels. MAIN OUTCOME MEASURES: The incidence of abnormal premature supraventricular depolarizations (SVPD) and the number of episodes of supraventricular tachycardia was defined as primary outcome measurements before the start of the study. In addition, heart rate oscillations (turbulence) after premature depolarizations and heart rate variability were compared at baseline and follow-up. RESULTS: SVPDs decreased from 59 +/- 29 to 21 +/- 8 per 24 h (P = 0.003), very early SVPDs (so called P on T) decreased from 36 +/- 24 to 3 +/- 1 per 24 h (P < 0.0001), respectively, and nonsustained supraventricular tachycardias decreased from 22 +/- 11 to 0.5 +/- 0.2 per 24 h (P = 0.01) after normalization of serum thyrotropin levels. The hyperthyroid phase was characterized by an increased heart rate (93 +/- 14 vs. 79 +/- 8 beats/min, P < 0.0001) and a decreased turbulence slope (3.6 vs. 9.2, P = 0.003), consistent with decreased vagal tone. This was confirmed by a significant decrease of heart rate variability. CONCLUSION: Hyperthyroidism is associated with an increased supraventricular ectopic activity in patients with normal hearts. The activation of these arrhythmogenic foci by elevated thyroid hormones may be an important causal link between hyperthyroidism and atrial fibrillation.

Ionic charge conservation and long-term steady state in the Luo-Rudy dynamic cell model.

It has been postulated that cardiac cell models accounting for changes in intracellular ion concentrations violate a conservation principle, and, as a result, computed parameters (e.g., ion concentrations and transmembrane potential, V(m)) drift in time, never attaining steady state. To address this issue, models have been proposed that invoke the charge conservation principle to calculate V(m) from ion concentrations ("algebraic" method), rather than from transmembrane current ("differential" method). The aims of this study are to compare model behavior during prolonged periods of pacing using the algebraic and differential methods, and to address the issue of model drift. We pace the Luo-Rudy dynamic model of a cardiac ventricular cell and compare the time-dependent behavior of computed parameters using the algebraic and differential methods. When ions carried by the stimulus current are taken into account, the algebraic and differential methods yield identical results and neither shows drift in computed parameters. The present study establishes the proper pacing protocol for simulation studies of cellular behavior during long periods of rapid pacing. Such studies are essential for mechanistic understanding of arrhythmogenesis, since cells are subjected to rapid periodic stimulation during many arrhythmias.

Mechanistic insights into very slow conduction in branching cardiac tissue: a model study.

It is known that branching strands of cardiac tissue can form a substrate for very slow conduction. The branches slow conduction by acting as current loads drawing depolarizing current from the main strand ("pull" effect). It has been suggested that, upon depolarization of the branches, they become current sources reinjecting current back into the strand, thus enhancing propagation safety ("push" effect). It was the aim of this study to verify this hypothesis and to assess the contribution of the push effect to propagation velocity and safety. Conduction was investigated in strands of Luo-Rudy dynamic model cells that branch from either a single branch point or from multiple successive branch points. In single-branching strands, blocking the push effect by not allowing current to flow retrogradely from the branches into the strand did not significantly increase the branching-induced local propagation delay. However, in multiple branching strands, blocking the push effect resulted in a significant slowing of overall conduction velocity or even in conduction failure. Furthermore, for certain slow velocities, the safety factor for propagation was higher when slow conduction was caused by branching tissue geometry than by reduced excitability without branching. Therefore, these results confirm the proposed "pull and push" mechanism of slow, but nevertheless robust, conduction in branching structures. Slow conduction based on this mechanism could occur in the atrioventricular node, where multiple branching is structurally present. It could also support reentrant excitation in diseased myocardium where the substrate is structurally complex.

Slow conduction in cardiac tissue: insights from optical mapping at the cellular level.

Under physiological conditions, slow conduction is essential for the function of the atrioventricular (AV) node, whereas, under pathophysiological conditions, slow conduction contributes importantly to the generation of life-threatening reentrant arrhythmias. This article addresses characteristics of slow conduction at the cellular network level during (a) a reduction of excitability, (b) a reduction of gap junctional coupling, and (c) in the setting of branching tissue structures. Microscopic impulse propagation in these settings was studied by using multiple site optical recording of transmembrane voltage in conjunction with patterned growth cultures of neonatal rat ventricular myocytes. In linear cell strands, a reduction of excitability slowed conduction by approximately 70% before block occurred. In contrast, critical reduction of gap junctional coupling induced a much higher degree of slowing (>99%) before block of conduction. Interestingly, a similar degree of conduction slowing was also observed in branching tissue structures under conditions of reduced excitability (98%). The finding of extremely slow but nevertheless safe conduction in these structures might be explained by a "pull and push" effect of the branches: by drawing electronic current from the activation wavefront, they first act as current loads which slow conduction at the branch points ("pull" effect). Then, on activation, they turn into current sources which feed current back into the system, thus supporting downstream activation and enhancing the safety of propagation ("push" effect). This "pull and push" mechanism may play a significant role in slow conduction in the AV node and in structurally discontinuous myocardium, such as the border regions of infarct scars.

Power-law behavior of beat-rate variability in monolayer cultures of neonatal rat ventricular myocytes.

It is known that extracardiac factors (nervous, humoral, and hemodynamic) participate in the power-law behavior of heart-rate variability. To assess whether intrinsic properties of cardiac tissue might also be involved, beat-rate variability was studied in spontaneously beating cell cultures devoid of extracardiac influences. Extracellular electrograms were recorded from monolayer cultures of neonatal rat ventricular myocytes under stable incubating conditions for up to 9 hours. The beat-rate time series of these recordings were examined in terms of their Fourier spectra and their Hurst scaling exponents. A non-0 Hurst exponent was found in 21 of 22 preparations (0.29+/-0.09; range, 0.11 to 0.45), indicating the presence of fractal self-similarity in the beat-rate time series. The same preparations exhibited power-law behavior of the power spectra with a power-law exponent of -1.36+/-0.24 (range, -1.04 to -1.96) in the frequency range of 0.001 to 1 Hz. Furthermore, it was found that the power-law exponent was nonstationary over time. These results indicate that the power-law behavior of heart-rate variability is determined not only by extracardiac influences but also by components intrinsic to cardiac tissue. Furthermore, the presence of power-law behavior in monolayer cultures of cardiomyocytes suggests that beat-rate variability might be determined by the complex nonlinear dynamics of processes occurring at the level of the cellular network, eg, interactions among a large number of cell oscillators or metabolic regulatory systems.

Optical recording of impulse propagation in designer cultures. Cardiac tissue architectures inducing ultra-slow conduction.

It has long been established that slow conduction constitutes one of the key mechanisms in the generation of cardiac arrhythmias. Also, it has been recognized that alterations in the cellular architecture of cardiac tissue can contribute to slow conduction. Based on the recent development of an experimental system permitting both the design of geometrically defined cardiac tissue structures in culture and the measurement of impulse propagation at the cellular level, we investigated the extent of conduction slowing along a tissue structure consisting of a cell strand releasing multiple side branches. This structure, which can functionally be looked upon as a series of interconnected current-to-load mismatches, gave rise to ultra-slow conduction (1-2 cm/s) that displayed a high margin of safety due to a "pull" and "push" effect exerted by the side branches on electrotonic current flow along the main strand. Under physiological conditions, such branching structures might contribute to slow conduction in the AV-node and, under pathophysiological conditions, to the precipitation of reentrant arrhythmias within minuscule tissue regions in a structurally remodeled myocardium. The results illustrate that the combination of patterned growth techniques and optical recording of transmembrane voltage are ideally suited to characterize systematically the relationship between tissue structure and impulse conduction.

Slow conduction in cardiac tissue, I: effects of a reduction of excitability versus a reduction of electrical coupling on microconduction.

It was the aim of this study to characterize the spread of activation at the cellular level in cardiac tissue during conduction slowing, a key element of reentrant arrhythmias; therefore, activation patterns were assessed at high spatiotemporal resolution in narrow (70 to 80 microm) and wide (230 to 270 microm) linear strands of cultured neonatal rat ventricular myocytes, using multiple site optical recording of transmembrane voltage. Slow conduction was induced by graded elevation of [K+]o, by applying tetrodotoxin, or by exposing the preparations to the gap junctional uncouplers palmitoleic acid or 1-octanol. The main findings of the study are 4-fold: (1) gap junctional uncoupling reduced conduction velocity (range, 37 to 47 cm/s under control conditions) to a substantially larger extent before block (

Slow conduction in cardiac tissue, II: effects of branching tissue geometry.

In cardiac tissue, functional or structural current-to-load mismatches can induce local slow conduction or conduction block, which are important determinants of reentrant arrhythmias. This study tested whether spatially repetitive mismatches result in a steady-state slowing of conduction. Patterned growth of neonatal rat heart cells in culture was used to design unbranched cell strands or strands releasing branches from either a single point or multiple points at periodic intervals. Electrical activation was followed optically using voltage-sensitive dyes under control conditions and in elevated [K+]o (5.8 and 14.8 mmol/L, respectively; in the latter case, propagation was carried by the L-type Ca2+ current). Preparations with multiple branch points exhibited discontinuous and slow conduction that became slower with increasing branch length and/or decreasing inter-branch distance. Compared with unbranched strands, conduction was maximally slowed by 63% under control conditions (from 44.9+/-3.4 to 16.7+/-1.0 cm/s) and by 93% in elevated [K+]o (from 15.7+/-2.3 to 1.1+/-0.2 cm/s). Local activation delays induced at a single branch point were significantly larger than the delays per branch point in multiple branching structures. Also, selective inactivation of inward currents in the branches induced conduction blocks. These 2 observations pointed to a dual role of the branches in propagation: whereas they acted as current sinks for the approaching activation thus slowing conduction ("pull" effect), they supplied, once excited, depolarizing current supporting downstream activation ("push" effect). This "pull and push" action resulted in a slowing of conduction in which the safety was largely preserved by the "push" effect. Thus, branching microarchitectures might contribute to slow conduction in tissue with discontinuous geometry, such as infarct scars and the atrioventricular node.

Optical recording system based on a fiber optic image conduit: assessment of microscopic activation patterns in cardiac tissue.

Optical recording of transmembrane voltage changes with the use of potentiometric dyes has opened the possibility of determining spatial patterns of electrical activity in excitable tissues. To follow such activation patterns on the cellular/subcellular level in heart cell cultures, a recording system was developed that features both high spatial resolution (4-200 microm) and high temporal resolution (uncertainty in the determination of delays between fast rising signals of +/-1 micros). Central to the system is a fiber optic image conduit consisting of 379 individual optical fibers. At one end the fibers are fused to form an input window that matches the size of the field of view of the microscope. At the other end, the fibers are loose, permitting a selectable subset to be connected to 80 discrete photodetectors. This design allows the sensitive area of the imager to be adapted to regions of interest in a given preparation, thus making optimal use of the limited number of detectors. Furthermore, by using a second fiber optic imager, individual photodetectors can be assigned to different optical ports, thus providing the means for fast and simultaneous dual-emission wavelength measurements. This feature permitted the elimination of motion artifacts arising from the myocytes without the use of contraction-suppressing drugs.

Paradoxical improvement of impulse conduction in cardiac tissue by partial cellular uncoupling.

Generally, impulse propagation in cardiac tissue is assumed to be impaired by a reduction of intercellular electrical coupling or by the presence of structural discontinuities. Contrary to this notion, the spatially uniform reduction of electrical coupling induced successful conduction in discontinuous cardiac tissue structures exhibiting unidirectional conduction block. This seemingly paradoxical finding can be explained by a nonsymmetric effect of uncoupling on the current source and the current sink in the preparations used. It suggests that partial cellular uncoupling might prevent the initiation of cardiac arrhythmias that are dependent on the presence of unidirectional conduction block.

Involvement of the calcium inward current in cardiac impulse propagation: induction of unidirectional conduction block by nifedipine and reversal by Bay K 8644.

In general, the fast sodium inward current (INa) is regarded as the main inward current ensuring fast and safe excitation of the normally polarized working myocardium. However, under conditions of locally delayed excitation in the millisecond range, the slow inward current (ICa) might additionally contribute to the success of impulse propagation. This hypothesis was tested in patterned growth cultures of neonatal rat ventricular myocytes, which consisted of narrow cell strands connected to large rectangular cell monolayers, where INa or ICa could be modified in the narrow cell strand adjacent to the expansion by a microsuperfusion system. As assessed during antegrade (strand-->expansion) propagation under control conditions using a system for multiple site optical recording of transmembrane voltage (MSORTV), this cell pattern gave either rise to local activation delays at the expansion ranging from 0.5 to 4 ms (dcontrol), or it induced undirectional conduction blocks (UCBs) in the antegrade direction. Irrespective of the size of dcontrol, suppression of the sodium current with tetrodotoxin confined to the cell strand adjacent to the expansion invariably induced UCB in the antegrade direction. If dcontrol was > 1 ms, UCB could also be elicited by suppression of ICa alone with nifedipine. Conversely, if UCB was present under control conditions, the inclusion of Bay K 8644 in the microsuperfusion established successful bidirectional conduction. These results suggest that ICa can be critically important for the success of impulse propagation across abrupt expansions of excitable tissue even if INa is not concurrently depressed.