toxin-ii-(anemonia-sulcata) and Disease-Models--Animal

Topics: Action Potentials; Animals; Cnidarian Venoms; Computer Simulation; Disease Models, Animal; Edema, Cardiac; Guinea Pigs; Heart Rate; Hypernatremia; Isolated Heart Preparation; Long QT Syndrome; Male; Models, Cardiovascular; Myocytes, Cardiac; NAV1.5 Voltage-Gated Sodium Channel; Sodium

Increased late sodium current (INa) induces long QT syndrome 3 with increased risk of atrial fibrillation (AF). The role of atrial late INa in the induction of AF and in the treatment of AF was determined in this study. AF parameters were measured in isolated rabbit hearts exposed to late INa enhancer and inhibitors. Late INa from isolated atrial and ventricular myocytes were measured using whole-cell patch-clamp techniques. We found that induced-AF by programmed S1S2 stimulation and spontaneous episodes of AF were recorded in hearts exposed to either low (0.1-3 nM) or high (3-10 nM) concentrations of ATX-II (n = 10). Prolongations in atrial monophasic action potential duration at 90% completion of repolarization and effective refractory period by ATX-II (0.1-15 nM) were greater in hearts paced at slow than at fast rates (n = 5-10, P < 0.05). Both endogenous and ATX-II-enhanced late INa density were greater in atrial than that in ventricular myocytes (n = 9 and 8, P < 0.05). Eleclazine and ranolazine reduced AF window and AF burden in association with the inhibition of both endogenous and enhanced atrial late INa with half maximal inhibitory concentrations (IC50) of 1.14 and 9.78, and 0.94 and 8.31 μM, respectively. The IC50s for eleclazine and ranolazine to inhibit peak INa were 20.67 and 101.79 μM, respectively, in atrial myocytes. In conclusion, enhanced late INa in atrial myocytes increases the susceptibility for AF. Inhibition of either endogenous or enhanced late INa, with increased atrial potency of drugs is feasible for the treatment of AF.

Topics: Action Potentials; Animals; Anti-Arrhythmia Agents; Atrial Fibrillation; Atrial Function; Cardiac Pacing, Artificial; Cnidarian Venoms; Disease Models, Animal; Female; Heart Atria; Heart Rate; Isolated Heart Preparation; Myocytes, Cardiac; Rabbits; Refractory Period, Electrophysiological; Sodium; Sodium Channel Blockers; Time Factors

Topics: Animals; Calcium; Cnidarian Venoms; Diastole; Disease Models, Animal; Echocardiography; Glycyrrhetinic Acid; Hemodynamics; Male; Microscopy, Confocal; Myocardial Reperfusion Injury; Random Allocation; Ranolazine; Rats; Tablets; Treatment Outcome

Female gender is a risk factor for long QT-related arrhythmias, but the underlying mechanisms remain uncertain. Here, we tested the hypothesis that gender-dependent function of the post-depolarization 'late' sodium current (I(Na-L)) contributes.. Studies were conducted in mice in which the canonical cardiac sodium channel Scn5a locus was disrupted, and expression of human wild-type SCN5A cDNA substituted. Baseline QT intervals were similar in male and female mice, but exposure to the sodium channel opener anemone toxin ATX-II elicited polymorphic ventricular tachycardia in 0/9 males vs. 6/9 females. Ventricular I(Na-L) and action potential durations were increased in myocytes isolated from female mice compared with those from males before and especially after treatment with ATX-II. Further, ATX-II elicited potentially arrhythmogenic early afterdepolarizations in myocytes from 0/5 male mice and 3/5 female mice.. These data identify variable late I(Na) as a modulator of gender-dependent arrhythmia susceptibility.

Topics: Acetanilides; Action Potentials; Animals; Cnidarian Venoms; Disease Models, Animal; Electrocardiography; Female; Genetic Predisposition to Disease; Humans; Long QT Syndrome; Male; Mice; Mice, 129 Strain; Mice, Inbred C57BL; Mice, Knockout; Mice, Transgenic; NAV1.5 Voltage-Gated Sodium Channel; Piperazines; Ranolazine; Risk Factors; Sex Factors; Tachycardia, Ventricular; Time Factors

There is clinical need to extend the understanding of epilepsy and to find novel approaches to treat this condition. Bang-sensitive (bs) Drosophila mutants, which exhibit reduced thresholds for seizure, offer an attractive possibility to combine tractable genetics, electrophysiology, and high-throughput screening. However, despite these advantages, the precise electrophysiological aberrations that contribute to seizure have not been identified in any bs mutant. Because of this, the applicability of Drosophila as a preclinical model has not yet been established. In this study, we show that electroshock of bs slamdance (sda) larvae was sufficient to induce extended seizure-like episodes. Whole cell voltage-clamp recordings from identified motoneurons (termed aCC and RP2) showed synaptic currents that were greatly increased in both amplitude and duration. Current-clamp recordings indicated that these inputs produced longer-lived plateau depolarizations and increased action potential firing in these cells. An analysis of voltage-gated currents in these motoneurons, in both first and third instar larvae, revealed a consistently increased persistent Na(+) current (I(Nap)) and a reduced Ca(2+) current in first instar larvae, which appeared normal in older third instar larvae. That increased I(Nap) may contribute to seizure-like activity is indicated by the observation that feeding sda larvae the antiepileptic drug phenytoin, which was sufficient to reduce I(Nap), rescued both seizure-like episode duration and synaptic excitation of motoneurons. In contrast, feeding of either anemone toxin, a drug that preferentially increases I(Nap), or phenytoin to wild-type larvae was sufficient to induce a bs behavioral phenotype. Finally, we show that feeding of phenytoin to gravid sda females was sufficient to both reduce I(Nap) and synaptic currents and rescue the bs phenotype in their larval progeny, indicating that a heightened predisposition to seizure may arise as a consequence of abnormal embryonic neural development.

Topics: Action Potentials; Animals; Anticonvulsants; Calcium Channels; Cnidarian Venoms; Disease Models, Animal; Drosophila melanogaster; Drosophila Proteins; Electroshock; Epilepsy, Reflex; Female; Genetic Predisposition to Disease; Larva; Motor Neurons; Mutation; Noise; Phenytoin; Seizures; Sodium Channels

Although long QT syndrome (LQTS) and coronary occlusion-reperfusion (O/R) are arrhythmogenic, they affect ventricular action potential duration (APD) differently. In contrast to the prolonged APD in LQTS, ischemia abbreviates APD after a transient prolongation. Thus we hypothesized that the dynamic interactive effects of ischemia and LQTS on APD and its dispersion would affect ventricular arrhythmogenicity. We mapped transmural distribution of action potentials in 6 groups of 10 isolated wedges of canine ventricular walls: LQTS-O/R, LQTS only, and O/R only, with separate groups for pacing cycle lengths (PCL) of 1,000 and 2,000 ms. We created type 3 LQTS with anemone toxin (ATX) II followed >30 min later by arterial occlusion (40 min) and reperfusion (>100 min). Arterial occlusion initially (first 4 min) prolonged and then shortened APD. Early afterdepolarizations (EADs) occurred during the initial 4 min of occlusion in 4 of the 10 LQTS-O/R wedges at PCL of 2,000 ms but not in the other groups. Reperfusion restored APD in the O/R-only groups but caused APD to overshoot its original duration, indicating depressed repolarization reserve, in the LQTS-O/R group. Reperfusion increased the dispersion of APDs and initiated ventricular tachycardia-fibrillation in 7 of 10 and 6 of 10 LQTS-O/R wedges and in 2 of 10 and 1 of 10 O/R-only wedges at PCLs of 1,000 and 2,000 ms, respectively. The LQTS-only wedges exhibited neither EADs nor ventricular tachycardia. We conclude that coronary O/R increased the arrhythmogenicity of LQTS via cumulative prolongation of APD, increase in repolarization dispersion, and suppression of repolarization reserve.

Topics: Action Potentials; Animals; Cnidarian Venoms; Coronary Disease; Disease Models, Animal; Dogs; In Vitro Techniques; Long QT Syndrome; Myocardial Reperfusion Injury; Reaction Time; Tachycardia, Ventricular

To define the cellular mechanisms responsible for the development of life-threatening arrhythmias in response to sympathetic activity in the congenital and acquired long QT syndromes (LCQTS).. Transmembrane action potentials (AP) from epicardial (EPI), M and endocardial (ENDO) cells and a transmural electrocardiogram were simultaneously recorded from an arterially perfused wedge of canine left ventricle. We examined the effect of beta-adrenergic agonists and antagonists on action potential duration (APD90), transmural dispersion of repolarization (TDR) and the development of Torsade de Pointes (TdP) in models of LQT1, LQT2 and LQT3 forms of LQTS.. I(Ks) block with chromanol 293B (LQT1) homogeneously prolonged APD90 of the three cell types without increasing TDR. Addition of isoproterenol prolonged QT and APD90 of M but abbreviated that of EPI and ENDO, causing a persistent increase in TDR; Torsade de Pointes developed or could be induced only in the presence of isoproterenol. I(Kr) block with d-sotalol (LQT2) and augmentation of late I(Na) with ATX-II (LQT3) prolonged APD90 of M more than EPI and ENDO, causing increases in QT and TDR. TdP developed in the absence of isoproterenol. In LQT2 isoproterenol initially prolonged, then abbreviated, the APD90 of M but always abbreviated EPI, thus transiently increasing TDR and the incidence of TdP. In LQT3, isoproterenol always abbreviated APD90 of the three cell types, causing a persistent decrease in TDR and suppression of TdP. The arrhythmogenic as well as protective actions of isoproterenol were reversed by propranolol.. Our data suggest that beta-adrenergic stimulation induces TdP by increasing transmural dispersion of repolarization in LQT1 and LQT2 but suppresses TdP by decreasing dispersion in LQT3. The data indicate that beta-blockers are protective in LQT1 and LQT2 but may facilitate TdP in LQT3.

Topics: Action Potentials; Adrenergic beta-Agonists; Adrenergic beta-Antagonists; Animals; Cardiotonic Agents; Chromans; Cnidarian Venoms; Disease Models, Animal; Dogs; Electrocardiography; Heart Conduction System; Heart Rate; Heart Ventricles; Isoproterenol; Long QT Syndrome; Neurotoxins; Potassium Channel Blockers; Propranolol; Sotalol; Sulfonamides; Sympathetic Nervous System; Treatment Outcome

The sea anemone toxin ATX II impairs skeletal muscle sodium channel inactivation, mimicking the persistent inward current observed in patients suffering from sodium channel myotonia. Mexiletine has beneficial effects on myotonia. To verify the efficiency of the drug on persistent inward current, we investigated the effect of 50 microM R(-)-mexiletine on sodium channels in cell-attached patches of rat skeletal muscle fibres, in the absence or presence of 2 microM ATX II. With the toxin, a proportion of channels displayed remarkable abnormal activity lasting the entire depolarisation, which resulted in a persistent inward current that represented up to 2.0% of the peak current. Mexiletine reduced by 75% the peak current elicited by depolarisation from -100 to -20 mV. This was due to the reduction by 60% of the maximal available peak current Imax and to the negative shift by -7 mV of steady-state inactivation. Mexiletine also greatly decreased the late current, but the effect was limited to 60% of reduction, comparable to that on Imax. Therefore mexiletine was able to block the ATX II-modified sodium channels, inhibiting the myotonia-producing persistent inward current.

Topics: Animals; Anti-Arrhythmia Agents; Cnidarian Venoms; Disease Models, Animal; In Vitro Techniques; Membrane Potentials; Mexiletine; Muscle Fibers, Skeletal; Muscle, Skeletal; Myotonia; Rats; Sodium Channels

1. Mutations that impair inactivation of the sodium channel in skeletal muscle have recently been postulated to cause several heritable forms of myotonia in man. A peptide toxin from Anemonia sulcata (ATX II) selectively disrupts the inactivation mechanism of sodium channels in a way that mimics these mutations. We applied ATX II to rat skeletal muscle to test the hypothesis that myotonia is inducible by altered sodium channel function. 2. Single-channel sodium currents were measured in blebs of surface membrane that arose from the mechanically disrupted fibres. ATX II impaired inactivation as demonstrated by persistent reopenings of sodium channels at strongly depolarized test potentials. A channel failed to inactivate, however, in only a small proportion of the depolarizing steps. With micromolar amounts of ATX II, the ensemble average open probability at the steady state was 0.01-0.02. 3. Ten micromolar ATX II slowed the relaxation of tension after a single twitch by an order of magnitude. Delayed relaxation is the in vitro analogue of the stiffness experienced by patients with myotonia. However, peak twitch force was not affected within the range of 0-10 microM ATX II. 4. Intracellular injection of a long-duration, constant current pulse elicited a train of action potentials in ATX II-treated fibres. After-depolarizations and repetitive firing often persisted beyond the duration of the stimulus. Trains of action potentials varied spontaneously in amplitude and firing frequency in a similar way to the electromyogram of a myotonic muscle. Both the after-depolarization and the post-stimulus firing were abolished by detubulating the fibres with glycerol. 5. We conclude that a loss of sodium channel inactivation alone, without changes in resting membrane conductance, is sufficient to produce the electrical and mechanical features of myotonia. Furthermore, in support of previous studies on myotonic muscle from patients, this model provides direct evidence that only a small proportion of sodium channels needs to function abnormally to cause myotonia.

Topics: Action Potentials; Animals; Cnidarian Venoms; Disease Models, Animal; Female; Humans; Hyperkalemia; In Vitro Techniques; Isometric Contraction; Muscles; Mutation; Myotonia; Paralyses, Familial Periodic; Rats; Sea Anemones; Sodium Channel Blockers; Sodium Channels