Dynamical mechanism for subcellular alternans in cardiac myocytes.

RATIONALE: Cardiac repolarization alternans is an arrhythmogenic rhythm disturbance, manifested in individual myocytes as a beat-to-beat alternation of action potential durations and intracellular calcium transient magnitudes. Recent experimental studies have reported "subcellular alternans," in which distinct regions of an individual cell are seen to have counterphase calcium alternations, but the mechanism by which this occurs is not well understood. Although previous theoretical work has proposed a possible dynamical mechanism for subcellular alternans formation, no direct evidence for this mechanism has been reported in vitro. Rather, experimental studies have generally invoked fixed subcellular heterogeneities in calcium-cycling characteristics as the mechanism of subcellular alternans formation. OBJECTIVE: In this study, we have generalized the previously proposed dynamical mechanism to predict a simple pacing algorithm by which subcellular alternans can be induced in isolated cardiac myocytes in the presence or absence of fixed subcellular heterogeneity. We aimed to verify this hypothesis using computational modeling and to confirm it experimentally in isolated cardiac myocytes. Furthermore, we hypothesized that this dynamical mechanism may account for previous reports of subcellular alternans seen in statically paced, intact tissue. METHODS AND RESULTS: Using a physiologically realistic computational model of a cardiac myocyte, we show that our predicted pacing algorithm induces subcellular alternans in a manner consistent with theoretical predictions. We then use a combination of real-time electrophysiology and fluorescent calcium imaging to implement this protocol experimentally and show that it robustly induces subcellular alternans in isolated guinea pig ventricular myocytes. Finally, we use computational modeling to demonstrate that subcellular alternans can indeed be dynamically induced during static pacing of 1D fibers of myocytes during tissue-level spatially discordant alternans. CONCLUSION: Here we provide the first direct experimental evidence that subcellular alternans can be dynamically induced in cardiac myocytes. This proposed mechanism may contribute to subcellular alternans formation in the intact heart.

Continuous and discontinuous radiofrequency energy delivery on the atrial free wall: Lesion transmurality, width, and biophysical characteristics.

BACKGROUND: Although lesion transmurality is required for durable pulmonary vein isolation, excess ablation is associated with increased risk of complications. OBJECTIVE: We sought to understand the impact of interrupted radiofrequency (RF) delivery conditions on lesion characteristics in the atrial free wall. METHODS: Thirty-three (11 left atrial, 22 right atrial) RF ablation lesions were created in the atria of 6 swine using power control mode (25 W, target contact force 15 g) with 1 of 3 conditions: 15 seconds ablation (n = 8), 30 seconds ablation (n = 14), or 2 15-second ablations at the same site separated by a 2-minute interruption (15 seconds × 2) (n = 11). RESULTS: Thirty of 33 lesions were transmural. Rates of transmurality (P = .45) and endocardial lesion width (5.6 ± 1.2 mm, P = .70) were similar between conditions. Mean tissue thickness was 1.7 ± 0.8 mm for transmural lesions. Wide variability in bipolar electrogram attenuation was observed across and within conditions and there were no significant between-group differences. Although impedance reductions were numerically greater in the 30-second and 15-second × 2 conditions (-14.6 ± 6.6 ohms and -14.0 ± 4.4 ohms, respectively) compared to the 15-second condition (-10.3 ± 6.4 ohms), variability was large, and differences were not statistically significant (P = .243). Impedance changes after ablation were largely transient. CONCLUSION: A single 15-second ablation at 25 W (target contact force of 15 g) with good stability produced similarly sized lesions compared to 30-second ablations and 2 15-second ablations at the same site in atrial free wall tissue. These data suggest over-ablation in the atria is common, larger-diameter lesions may require greater power, and many clinically available parameters of lesion size may be unreliable on the posterior wall.

Feedback-control induced pattern formation in cardiac myocytes: a mathematical modeling study.

Cardiac alternans is a dangerous rhythm disturbance of the heart, in which rapid stimulation elicits a beat-to-beat alternation in the action potential duration (APD) and calcium (Ca) transient amplitude of individual myocytes. Recently, "subcellular alternans", in which the Ca transients of adjacent regions within individual myocytes alternate out-of-phase, has been observed. A previous theoretical study suggested that subcellular alternans may result during static pacing from a Turing-type symmetry breaking instability, but this was only predicted in a subset of cardiac myocytes (with negative Ca to voltage (Ca-->V(m)) coupling) and has never been directly verified experimentally. A recent experimental study, however, showed that subcellular alternans is dynamically induced in the remaining subset of myocytes during pacing with a simple feedback control algorithm ("alternans control"). Here we show that alternans control pacing changes the effective coupling between the APD and the Ca transient (V(m)-->Ca coupling), such that subcellular alternans is predicted to occur by a Turing instability in cells with positive Ca-->V(m) coupling. In addition to strengthening the understanding of the proposed mechanism for subcellular alternans formation, this work (in concert with previous theoretical and experimental results) illuminates subcellular alternans as a striking example of a biological Turing instability in which the diffusing morphogens can be clearly identified.

Impact of interruptions in radiofrequency energy delivery on lesion characteristics.

BACKGROUND: During catheter ablation, delivery of radiofrequency (RF) energy to a target site is sometimes interrupted by catheter instability and clinical factors. The impact of interruption of RF delivery on lesion characteristics has not been characterized. OBJECTIVE: The purpose of this study was to determine the impact of interruption of RF application on lesion size. METHODS: Forty-two RF ablation lesions (21 left ventricle, 21 right ventricle) were created in the ventricles of 6 swine using power control mode (30 W; target contact force 15g) with 1 of 3 conditions: 15-second ablation (15s), 30-second ablation (30s), or two 15-second ablations (15s×2) at the same site separated by a 2-minute pause. RESULTS: Lesion volume was significantly larger for 30s lesions (501 ± 146 mm3) compared to both 15s×2 (314 ± 98 mm3) and 15s (242 ± 104 mm3) lesions (P <.001 for both pairwise comparisons). Compared to 15s lesions, lesion volume was numerically greater for 15s×2 lesions, but this did not reach statistical significance (P = .087). Differences in lesion volume between 30s and 15s×2 lesions were driven mainly by differences in lesion width (10.7 ± 1.1 mm vs 9.1 ± 1.7 mm; P = .04) rather than depth (9 ± 1.2 mm vs 8.4 ± 1.2 mm; P = .29). There were no differences in mean contact force by group. There was no difference in total force-time integral for the 30s and 15s×2 lesion groups [median 444 (interquartile range 312) g∙s vs 380 (164) g∙s; P = 1]. CONCLUSION: Compared to lesions resulting from continuous RF ablation, lesions resulting from interrupted ablation have a smaller overall lesion volume, predominantly due to smaller lesion width. These data suggest that if disruption in energy delivery occurs, lesions may need closer spacing to avoid gaps.

Extra-cardiac manifestations of adult congenital heart disease.

Advancement in correction or palliation of congenital cardiac lesions has greatly improved the lifespan of congenital heart disease patients, resulting in a rapidly growing adult congenital heart disease (ACHD) population. As this group has increased in number and age, emerging science has highlighted the systemic nature of ACHD. Providers caring for these patients are tasked with long-term management of multiple neurologic, pulmonary, hepatic, renal, and endocrine manifestations that arise as syndromic associations with congenital heart defects or as sequelae of primary structural or hemodynamic abnormalities. In this review, we outline the current understanding and recent research into these extra-cardiac manifestations.

Non-linear dynamics of cardiac alternans: subcellular to tissue-level mechanisms of arrhythmia.

Cardiac repolarization alternans is a rhythm disturbance of the heart in which rapid stimulation elicits a beat-to-beat alternation in the duration of action potentials and magnitude of intracellular calcium transients in individual cardiac myocytes. Although this phenomenon has been identified as a potential precursor to dangerous reentrant arrhythmias and sudden cardiac death, significant uncertainty remains regarding its mechanism and no clinically practical means of halting its occurrence or progression currently exists. Cardiac alternans has well-characterized tissue, cellular, and subcellular manifestations, the mechanisms and interplay of which are an active area of research.

MinK-dependent internalization of the IKs potassium channel.

AIMS: KCNQ1-MinK potassium channel complexes (4alpha:2beta stoichiometry) generate IKs, the slowly activating human cardiac ventricular repolarization current. The MinK ancillary subunit slows KCNQ1 activation, eliminates its inactivation, and increases its unitary conductance. However, KCNQ1 transcripts outnumber MinK transcripts five to one in human ventricles, suggesting KCNQ1 also forms other heteromeric or even homomeric channels there. Mechanisms governing which channel types prevail have not previously been reported, despite their significance: normal cardiac rhythm requires tight control of IKs density and kinetics, and inherited mutations in KCNQ1 and MinK can cause ventricular fibrillation and sudden death. Here, we describe a novel mechanism for this control. METHODS AND RESULTS: Whole-cell patch-clamping, confocal immunofluorescence microscopy, antibody feeding, biotin feeding, fluorescent transferrin feeding, and protein biochemistry techniques were applied to COS-7 cells heterologously expressing KCNQ1 with wild-type or mutant MinK and dynamin 2 and to native IKs channels in guinea-pig myocytes. KCNQ1-MinK complexes, but not homomeric KCNQ1 channels, were found to undergo clathrin- and dynamin 2-dependent internalization (DDI). Three sites on the MinK intracellular C-terminus were, in concert, necessary and sufficient for DDI. Gating kinetics and sensitivity to XE991 indicated that DDI decreased cell-surface KCNQ1-MinK channels relative to homomeric KCNQ1, decreasing whole-cell current but increasing net activation rate; inhibiting DDI did the reverse. CONCLUSION: The data redefine MinK as an endocytic chaperone for KCNQ1 and present a dynamic mechanism for controlling net surface Kv channel subunit composition-and thus current density and gating kinetics-that may also apply to other alpha-beta type Kv channel complexes.

Gene expression analysis in mice with elevated glial fibrillary acidic protein and Rosenthal fibers reveals a stress response followed by glial activation and neuronal dysfunction.

Alexander disease is a fatal neurodegenerative disorder resulting from missense mutations of the intermediate filament protein, GFAP. The pathological hallmark of this disease is the formation of cytoplasmic protein aggregates within astrocytes known as Rosenthal fibers. Transgenic mice engineered to over-express wild-type human GFAP develop an encephalopathy with identical aggregates, suggesting that elevated levels of GFAP in addition to mutant protein contribute to the pathogenesis of this disorder. To study further the effects of elevated GFAP and Rosenthal fibers per se, independent of mutations, we performed gene expression analysis on olfactory bulbs of transgenic mice at two different ages to follow the progression of pathology. The expression profiles reveal a stress response that includes genes involved in glutathione metabolism, peroxide detoxification and iron homeostasis. Many of these genes are regulated by the transcription factor Nfe2l2, which is also increased in expression at 3 weeks. An immune-related response occurs with activation of cytokine and cytokine receptor genes, complement components and acute phase response genes. These transcripts are further elevated with age, with additional induction of macrophage-specific markers such as Mac1 and CD68, suggesting activation of microglia. At 4 months, decreased expression of genes for microtubule-associated proteins, vesicular trafficking proteins and neurotransmitter receptors becomes apparent. Interneuron-specific transcription factors including Dlx family members and Pax6 are downregulated as well as Gad1 and Gad2, suggesting impairment of GABAergic granule cells. Together, these data implicate an initial stress response by astrocytes, which results in the activation of microglia and compromised neuronal function.