Action Potential Configuration Research Articles

Effect of ethanol on action potential and ionic membrane currents in rat ventricular myocytes

Even though alcohol intoxication is often linked to arrhythmias, data describing ethanol effect on cardiac ionic channels are rare. In addition, ethanol is used as a solvent of hydrophobic compounds in experimental studies. We investigated changes of the action potential (AP) configuration and main ionic membrane currents in rat cardiomyocytes under 20-1500 m(M) ethanol. Experiments were performed on enzymatically isolated rat right ventricular myocytes using the whole cell patch-clamp technique at room temperature. Ethanol reversibly decelerated the upstroke velocity and decreased AP amplitude and duration at 0.2 and 3 Hz. The fast sodium current I(Na) , l-type calcium current I(Ca) and transient outward potassium current I(to) were reversibly inhibited in a concentration-dependent manner (50% inhibition at 446 ± 12, 553 ± 49 and 1954 ± 234 m(M), respectively, with corresponding Hill coefficients 3.1 ± 0.3, 1.1 ± 0.2 and 0.9 ± 0.1). Suppression of I(Na) and I(Ca) magnitude was slightly voltage dependent. The effect on I(Ca) and I(to) was manifested mainly as an acceleration of their apparent inactivations with a decreased slow and fast time constant respectively. As a consequence of marked differences in n(H) , sensitivity of the currents to ethanol at 10% inhibition decreases in the following order: I(Ca) (75 mm, 3.5‰), I(to) (170 m(M), 7.8‰) and I(Na) (220 m(M), 10.1‰). Our results suggest a slight inhibition of all the currents at ethanol concentrations relevant to deep alcohol intoxication. The concentration dependence measured over a wide range may serve as a guideline when using ethanol as a solvent.

Control of Ca2+ Release by Action Potential Configuration in Normal and Failing Murine Cardiomyocytes

Cardiomyocytes from failing hearts exhibit spatially nonuniform or dyssynchronous sarcoplasmic reticulum (SR) Ca2+ release. We investigated the contribution of action potential (AP) prolongation in mice with congestive heart failure (CHF) after myocardial infarction. AP recordings from CHF and control myocytes were included in a computational model of the dyad, which predicted more dyssynchronous ryanodine receptor opening during stimulation with the CHF AP. This prediction was confirmed in cardiomyocyte experiments, when cells were alternately stimulated by control and CHF AP voltage-clamp waveforms. However, when a train of like APs was used as the voltage stimulus, the control and CHF AP produced a similar Ca2+ release pattern. In this steady-state condition, greater integrated Ca2+ entry during the CHF AP lead to increased SR Ca2+ content. A resulting increase in ryanodine receptor sensitivity synchronized SR Ca2+ release in the mathematical model, thus offsetting the desynchronizing effects of reduced driving force for Ca2+ entry. A modest nondyssynchronous prolongation of Ca2+ release was nevertheless observed during the steady-state CHF AP, which contributed to increased time-to-peak measurements for Ca2+ transients in failing cells. Thus, dyssynchronous Ca2+ release in failing mouse myocytes does not result from electrical remodeling, but rather other alterations such as T-tubule reorganization.

High-mobility group box 1 (HMGB1) downregulates cardiac transient outward potassium current ( Ito) through downregulation of Kv4.2 and Kv4.3 channel transcripts and proteins

Transient outward potassium currents ( I to) are major early repolarization currents in shaping cardiac action potential (AP). Downregulation of I to contributes to AP configuration alteration in myocardial infarction (MI) and numerous other heart diseases. High-mobility group box 1 (HMGB1), a proinflammatory cytokine, has been reported to increase dramatically in the serum of patients with MI, participating in ischemia–reperfusion injury and recovery of post-infarction failing heart. This study investigated the possible role of HMGB1 in regulating cardiac I to and electrical stability. HMGB1 treatment for 24 h significantly inhibited the current densities of heterologously expressed Kv4.3 and Kv4.2 in COS-7 cells and native I to in neonatal rat ventricular myocytes (NRVMs) in a dose-dependent manner. HMGB1 decreased the mRNA and protein levels of the I to α subunits Kv4.2 and Kv4.3 channels, but not the β subunit KChIP2 and KCNE2 in NRVMs. The receptor binding domain (150–186 amino acid residues) responsible for receptor of advanced glycation end product (RAGE) binding similarly inhibited I to , while treatment with soluble RAGE that blocks binding of ligands to cell-surface RAGE partially restored I to current density and Kv4 protein expressions. Box A which possesses no proinflammatory activity of HMGB1 still remained part of the I to suppression effect. In addition to downregulating I to, HMGB1 modestly inhibited L-type Ca 2+ current, but not I K1. The AP duration (APD) was slightly prolonged by HMGB1 treatment. These results collectively establish HMGB1 as a novel pathological factor downregulating I to partially through HMGB1–RAGE interaction, providing new insights into the potential molecular mechanisms underlying the electrical remodeling in MI.

Non‐quantal release of acetylcholine from parasympathetic nerve terminals in the right atrium of rats

Acetylcholinesterase (AChE) inhibitors provoke typical cholinergic effects in the isolated right atrium of the rat due to the accumulation of acetylcholine (ACh). Our study was designed to show that in the absence of vagal impulse activity, ACh is released from the parasympathetic nerve fibres by means of non-quantal secretion. The conventional microelectrode technique was used to study changes in action potential (AP) configuration in the right atrium preparation of rats during application of AChE inhibitors. Staining with the lipophilic fluorescent dye FM1-43 was used to demonstrate the presence of endocytosis in cholinergic endings. The AChE inhibitors armin (10(7)-10(5)m) and neostigmine (10(7) to 5 x 10(6)m) caused a reduction of AP duration and prolonged the cycle length. These effects were abolished by atropine and were therefore mediated by ACh accumulated in the myocardium during AChE inhibition. Putative block of impulse activity of the postganglionic neurons by tetrodotoxin (5 x 10(7)m) and blockade of ganglionic transmission by hexomethonium (2 x 10(4)m), as well as blockade of all forms of quantal release with Clostridium botulinum type A toxin (50 U ml(1)), did not alter the effects of armin. Experiments with FM1-43 dye confirmed the effective block of exocytosis by botulinum toxin. Selective inhibition of the choline uptake system using hemicholinium III (10(5)m), which blocks non-quantal release at the neuromuscular junction, suppressed the effects of AChE inhibitors. Thus, accumulation of ACh is likely to be caused by non-quantal release from cholinergic terminals. We propose that non-quantal release of ACh, shown previously at the neuromuscular junction, is present in cholinergic postganglionic fibres of the rat heart in addition to quantal release.

Control of Ca Release Synchrony by Action Potential Configuration in Murine Cardiomyocytes

Spatially non-uniform or “dyssynchronous” sarcoplasmic reticulum (SR) Ca release has been reported in cardiomyocytes from failing hearts. Using a murine model of congestive heart failure (CHF) following myocardial infarction, we investigated whether altered action potential (AP) configuration promotes release dyssynchrony. We observed that APs (1 Hz) were prolonged in cardiomyocytes isolated from the viable septum of CHF hearts, compared to and sham-operated controls (SHAM). Representative AP recordings were included in a detailed computational model of the Ca dynamics in the dyad. The model predicted reduced driving force for L-type Ca current and more dyssynchronous opening of ryanodine receptors during stimulation with the CHF AP than the SHAM AP. These predictions were confirmed in isolated cardiomyocyte experiments, when cells were alternately stimulated by SHAM and CHF AP voltage-clamp waveforms. However, when a train of like APs was used as the voltage stimulus, the SHAM and CHF AP produced a similar Ca release pattern. In this steady-state condition, both modeling and cell experiments showed that greater integrated Ca entry during the CHF AP lead to increased SR Ca content. We modeled the effect of increased SR Ca content by increasing the Ca sensitivity of the ryanodine receptor, which we observed increased the synchrony of ryanodine receptor activation. Thus, at steady-state, Ca release synchrony was maintained during the CHF AP as greater ryanodine sensitivity offset the de-synchronizing effects of reduced driving force for Ca entry. Our results suggest that dyssynchronous Ca release in failing mouse myocytes results from alterations such as T-tubule re-organization, and not electrical remodeling.

KCNH2 Channel Activators Increase Ikr in Hl-1 Cardiomyocytes and May Prevent the Occurrence of Torsades De Pointes in Long QT Syndrome

Introduction: Several KCNH2 channel activators may provide a novel pharmacological approach for the treatment of long QT syndrome (LQTS). We therefore investigated the effects of the activators on IKr and action potential of cardiac myocytes.Methods: We characterized the actions of three KCNH2 channel activators, mallotoxin (MTX), PD-118057 (PD), and NS1643 (NS) on IKr of HL-1 cardiomyocytes, using the whole cell patch clamp technique. With a mathematical model of human ventricular myocytes, we further evaluated the impact of activator-induced changes in IKr kinetics on the action potential configuration in normal and LQTS.Results: The maximum tail currents of IKr were 23.4±3.5 nA/pF with 10 μM MTX (n=12), 22.1±2.7 nA/pF with 10 μM PD (n=13), and 23.3±2.7 nA/pF with 10 μM NS (n=16), which were significantly greater than 12.8±1.0 nA/pF in control (n=38). The half-maximal activation voltage was significantly shifted from −1.8±2.7 (n=38) to −13.0±2.3 (n=11), −8.3±2.1 (n=13), and −14.7±3.2 (n=14) mV by MTX, PD, and NS, respectively. Deactivation during the repolarization to −40 mV was significantly slowed by MTX, but not by PD or NS. The half-maximal inactivation voltage was significantly shifted from −6.6±2.2 (n=28) to −29.9±2.9 (n=15) mV by MTX, but not by PD, and NS. Simulation study showed that the activator-induced changes of IKr increased the amplitude of IKr during phase 2 of action potentials and consequently shortened the action potential duration by 19.7-23.6% in LQT1 and LQT3 models. A reduction of IKr in the LQT3 model evoked early afterdepolarization, which was abolished by the activator-induced enhancement of IKr.Conclusion: KCNH2 channel activators, mallotoxin, PD-118057, and NS1643 increases IKr through distinct kinetic mechanisms and can be utilized for potential therapy of LQTS and torsades de pointes.

Changes in Aβ Non-Nociceptive Primary Sensory Neurons in a Rat Model of Osteoarthritis Pain

BackgroundPain is a major debilitating factor in osteoarthritis (OA), yet few mechanism-based therapies are available. To address the need to understand underlying mechanisms the aim of the present study was to determine changes in sensory neurons in an animal model of OA pain.ResultsThe model displayed typical osteoarthritis pathology characterized by cartilage degeneration in the knee joint and also manifested knee pathophysiology (edema and increased vasculature permeability of the joint) and altered nociception of the affected limb (hind paw tenderness and knee articulation-evoked reduction in the tail flick latency). Neurons included in this report innervated regions throughout the entire hind limb. Aβ-fiber low threshold mechanoreceptors exhibited a slowing of the dynamics of action potential (AP) genesis, including wider AP duration and slower maximum rising rate, and muscle spindle neurons were the most affected subgroup. Only minor AP configuration changes were observed in either C- or Aδ-fiber nociceptors.ConclusionThus, at one month after induction of the OA model Aβ-fiber low threshold mechanoreceptors but not C- or Aδ-fiber nociceptors had undergone changes in electrophysiological properties. If these changes reflect a change in functional role of these neurons in primary afferent sensory processing, then Aβ-fiber non-nociceptive primary sensory neurons may be involved in the pathogenesis of OA pain. Further, it is important to point out that the patterns of the changes we observed are consistent with observations in models of peripheral neuropathy but not models of peripheral inflammation.

Contributions of Ion Channel Currents to Ventricular Action Potential Changes and Induction of Early Afterdepolarizations During Acute Hypoxia

Variability in delivery of oxygen can lead to electric instability in the myocardium and the generation of arrhythmias. In addition ischemic heart disease and angina are associated with an increase in circulating catecholamines that further increases the risk of developing ventricular tachyarrhythmias. We investigated the net effects of acute hypoxia and catecholamines on the cardiac action potential. We incorporated all published data on the effects of hypoxia on the late Na(+) current (I(Na-L)), the fast Na(+) current (I(Na)), the basal L-type Ca(2+) channel current (I(Ca-L)), and the slow (I(Ks)) and rapid components of the delayed rectifier K(+)-current (I(Kr)) in the absence and presence of beta-adrenergic receptor (beta-AR) stimulation into the Luo-Rudy model of the action potential. Hypoxia alone had little effect on the action potential configuration or action potential duration. However in the presence of beta-AR stimulation, hypoxia caused a prolongation of the action potential and early afterdepolarizations (EADs) and spontaneous tachycardia were induced. Experiments performed in guinea pig ventricular myocytes confirmed the modeling results. EADs occur predominantly because of the increased sensitivity of I(Ca-L) to beta-AR stimulation during hypoxia. beta-AR stimulation is necessary to induce EADs as EADs are never observed during hypoxia in the absence of beta-AR stimulation.

The Effect of Hydrogen Sulfide on Electrical Activity of Rat Atrial Myocardium

Changes in the configuration of action potentials and in the frequency of pacemaker discharges in a preparation of isolated rat right atrium under the effect of sodium hydrosulfide degrading in water solution with hydrogen sulfide release were studied by intracellular recording of action potentials in the myocardium. Sodium hydrosulfide in concentrations of 100-500 microM markedly reduced the duration of action potentials at the level of 50 and 90% repolarization and decelerated the sinus rhythm. Moreover, sodium hydrosulfide shortened action potentials in the preparations working in the forced rhythm. Glybenclamide (potassium ATP-dependent channel blocker; 10 microM) reduced the effect sodium hydrosulfide (200 microM) by more than 60%, which suggested the involvement of potassium ATP-dependent current in the realization of the effect of hydrogen sulfide on configuration of action potentials. Hence, hydrogen sulfide, recently described as a signal compound, modulates many electrophysiological parameters of the myocardium.

Self‐augmentation of the lengthening of repolarization is related to the shape of the cardiac action potential: implications for reverse rate dependency

The aims of the present work were to study the mechanism of the reverse rate dependency of different interventions prolonging cardiac action potential duration (APD). The reverse rate-dependent lengthening effect of APD-prolonging interventions and the possible involvement of I(Kr) (rapid component of the delayed rectifier potassium current) and I(K1) (inward rectifier potassium current) were studied by using the standard microelectrode and the whole-cell patch-clamp techniques in dog multicellular ventricular preparations and in myocytes isolated from undiseased human and dog hearts. All applied drugs--dofetilide (1 micromol.L(-1)), BaCl(2) (10 micromol.L(-1)), BAY-K-8644 (1 micromol.L(-1)), veratrine (1 microg.mL(-1))--lengthened APD in a reverse rate-dependent manner regardless of their mode of action, suggesting that reverse rate dependency may not represent a specific mechanism of APD prolongation. The E-4031-sensitive current (I(Kr)) and the Ba(2+)-sensitive current (I(K1)) were recorded during repolarizing voltage ramps having various steepness and also during action potential waveforms with progressively prolonged APD. Gradually delaying repolarization results in smaller magnitude of I(Kr) and I(K1) currents at an isochronal phase of the pulses. This represents a positive feedback mechanism, which appears to contribute to the reverse rate-dependent prolongation of action potentials. Action potential configuration may influence the reverse rate-dependent APD prolongation due to the intrinsic properties of I(Kr) and I(K1) currents. Drugs lengthening repolarization by decreasing repolarizing outward, or increasing depolarizing inward, currents are expected to cause reverse rate-dependent APD lengthening with high probability, regardless of which current they modify.

Mechanisms of Cardiac Muscle Insensitivity to a Novel Acetylcholinesterase Inhibitor C-547

We compared the effects of the novel acetylcholinesterase (AChE) inhibitor C-547 on action potential configuration and sinus rhythm in the isolated right atrium preparation of rat with those of armin and neostigmine. Both armin (10(-7), 10(-6), and 10(-5) M) and neostigmine (10(-7), 10(-6), and 5 x 10(-6) M) produced a marked decrease in action potential duration and slowing of sinus rate. These effects were abolished by atropine and are attributable to the accumulation of acetylcholine in the myocardium. The novel selective AChE inhibitor C-547 (10(-9) to 10(-7) M), an alkylammonium derivative of 6-methyluracil, had no such effects. The inhibition constant of C-547 on cardiac AChE is 40-fold higher than that on extensor digitorum longus muscle AChE. These results suggest that C-547 might be employed to treat diseases such as myasthenia gravis or Alzheimer disease, without having unwanted effects on the heart.

Effect of Short-term Cardiac Memory on Ventricular Electrical Restitution and QT Intervals in Humans

Cardiac memory (CM) can alter the configuration of action potentials and the transmural repolarization gradient in ventricular tissue. This study evaluated the effects of CM on ventricular arrhythmogenicity. A total of 20 patients (12 females, 8 males; mean age, 46 +/- 13 years) were enrolled. The following indicators were measured to evaluate ventricular arrhythmogenicity: (1) the action potential duration at 90% repolarization (APD90) recorded from the right ventricular apex (RVA); (2) the maximal slope of the action potential duration restitution curve (APDR) constructed by programmed extra stimuli from RVA; and (3) the maximal corrected QT interval (QTc) and QT interval dispersion (QTd). The short-term CM was induced by constant pacing from the RVA at a pacing cycle length (PCL) of 400 ms for 20 minutes. After induction of CM, the mean APD90 were significantly shortened at both PCLs of 600 ms and 400 ms (252.9 +/- 6.4 ms vs. 235.6 +/- 6.4 ms and 231.2 +/- 6.4 ms vs. 214.4 +/- 7.3 ms, respectively; p = 0.001). No significant change regarding the maximal slopes of APDR were found at both PCLs of 600 ms and 400 ms (1.05 +/- 0.09 vs. 0.96 +/- 0.11 and 0.85 +/- 0.08 vs. 0.84 +/- 0.09, respectively). QTc (417.3 +/- 9.1 ms vs. 454.7 +/- 8.3 ms; p = 0.001), but not QTd (63.4 +/- 5.4 ms vs. 65.7 +/- 6.1 ms), was significantly shortened. Short-term CM significantly decreased ventricular APD90 and QTc, but did not significantly change the maximal slope of APDR or QTd. These results suggest that CM might not have a significant effect on ventricular arrhythmogenicity.

Deleting the accessory subunit KChIP2 results in loss of Ito,f and increased IK,slow that maintains normal action potential configuration

Four voltage-gated potassium currents, I(to,f) (K(V)4.2), I(to,s) (K(V)1.4), I(K,slow) (K(V)1.5+K(V)2.1), and I(SS) (TASK1), govern murine ventricular repolarization. Although the accessory subunit KChIP2 influences I(to,f) expression, in preliminary experiments we found that action potential duration (APD) is maintained in KChIP2 knockout mice. We tested the role of KChIP2 in regulating APD and studied the underlying ionic currents. We used microelectrode techniques, whole-cell patch clamp studies, and real-time polymerase chain reaction amplification to characterize ventricular repolarization and its determinants in wild-type and KChIP2(-/-) mice. Despite comparable baseline action potentials, APD was more markedly prolonged by 4-aminopyridine (4-AP) in KChIP2(-/-) preparations. Peak K(+) current densities were similar in wild-type and KChIP2(-/-) cells (mean +/- SEM I(P): 28.3 +/- 2 (n = 27) vs. 29.2 +/- 2 pA/pF (n = 24), respectively; P > .05). Heteropodatoxin-2 (HpTx-2, 1 microM) had no effect on current amplitude in KChIP2(-/-) myocytes. The current fractions sensitive to 4-AP (50 microM and 1 mM) were larger in KChIP2(-/-) than wild-type (P < .05). Real-time polymerase chain reaction showed absence of KChIP2 and increased K(V)1.5 expression in KChIP2(-/-) ventricular myocardium. KChIP2 deficiency eliminated HpTx-2-sensitive I(to,f), but had little impact on total APD, secondary to upregulation of 4-AP-sensitive I(K,slow) in association with increased K(V)1.5 expression. There is increased sensitivity to 4-AP-mediated APD prolongation in KChIP2(-/-). Thus, KChIP2 seems important for murine repolarization in circumstances of reduced repolarization reserve.

Chronic myocardial infarction induces phenotypic and functional remodeling in the guinea pig cardiac plexus

Chronic myocardial infarction (CMI) is associated with remodeling of the ventricle and evokes adaption in the cardiac neurohumoral control systems. To evaluate the remodeling of the intrinsic cardiac nervous system following myocardial infarction, the dorsal descending coronary artery was ligated in the guinea pig heart and the animals were allowed to recover for 7-9 wk. Thereafter, atrial neurons of the intrinsic cardiac plexus were isolated for electrophysiological and immunohistochemical analyses. Intracellular voltage recordings from intrinsic cardiac neurons demonstrated no significant changes in passive membrane properties or action potential configuration compared with age-matched controls and sham-operated animals. The intrinsic cardiac neurons from chronic infarcted hearts did demonstrate an increase in evoked action potential (AP) frequency (as determined by the number of APs produced with depolarizing stimuli) and an increase in responses to exogenously applied histamine compared with sham and age-matched controls. Conversely, pituitary adenylate cyclase-activating polypeptide (PACAP)-induced increases in intrinsic cardiac neuron-evoked AP frequency were similar between control and CMI animals. Immunohistochemical analysis demonstrated a threefold increase in percentage of neurons immunoreactive for neuronal nitric oxide synthase (NOS) in CMI animals compared with control and the additional expression of inducible NOS by some neurons, which was not evident in control animals. Finally, the density of mast cells within the intrinsic cardiac plexus was increased threefold in preparations from CMI animals. These results indicate that CMI induces a differential remodeling of intrinsic cardiac neurons and functional upregulation of neuronal responsiveness to specific neuromodulators.

The fruit essential oil of Pimpinella anisum L. (Umblliferae) induces neuronal hyperexcitability in snail partly through attenuation of after-hyperpolarization

Aim of the study Many biological actions of Pimpinella anisum L. (Ainse), including antiepileptic activity have been demonstrated; however, there is no data concerning its precise cellular mechanisms of action. We determined whether the fruit essential oil of anise affect the bioelectrical activity of snail neurons in control condition or after pentylenetetrazol (PTZ) induced epileptic activity. Materials and methods Intracellular recordings were made under the current clamp condition and the effects of anise oil (0.01% or 0.05%) alone or in combination with PTZ were assessed on the firing pattern, action potential configuration and postspike potentials. Results Anise oil changed the firing pattern from regular tonic discharge to irregular and then to bursting in intact cells or resulted in the robustness of the burst firing and the steepness of the paroxysmal shift induced by PTZ treatment. It also significantly increased the firing frequency and decreased both the after-hyperpolarization potential (AHP) following single action potential and the post-pulse AHP. Conclusions Likely candidate cellular mechanisms underlying the hyperexcitability produced by anise oil include enhancement of Ca 2+ channels activity or inhibition of voltage and/or Ca 2+ dependent K + channels activity underlying AHPs. These finding indicates that a certain caution is needed when Pimpinella anisum is used for treating patients suffering from epilepsy.

Contribution of I Kr and I K1 to ventricular repolarization in canine and human myocytes: is there any influence of action potential duration?

The aim of the present work was to study the profile of the rapid delayed rectifier potassium current (I (Kr)) and the inward rectifier potassium current (I (K1)) during ventricular repolarization as a function of action potential duration and rate of repolarization. Whole cell configuration of the patch clamp technique was used to monitor I (Kr) and I (K1) during the action potential plateau and terminal repolarization. Action potentials recorded at various cycle lengths (0.4-5 s) and repolarizing voltage ramps having various slopes (0.5-3 V/s) were used as command signals. I (Kr) and I (K1) were identified as difference currents dissected by E-4031 and BaCl(2), respectively. Neither peak amplitudes nor mean values of I (Kr) and I (K1) recorded during the plateau of canine action potentials were influenced by action potential duration. The membrane potential where I (Kr) and I (K1) peaked during the terminal repolarization was also independent of action potential duration. Similar results were obtained in undiseased human ventricular myocytes, and also in canine cells when I (Kr) and I (K1) were evoked using repolarizing voltage ramps of various slopes. Action potential voltage clamp experiments revealed that the peak values of I (Kr), I (K1), and net outward current during the terminal repolarization were independent of the pacing cycle length within the range of 0.4 and 5 s. The results indicate that action potential configuration fails to influence the amplitude of I (Kr) and I (K1) during the ventricular action potential in dogs and humans, suggesting that rate-dependent changes in action potential duration are not likely related to rate-dependent alterations in I (Kr) or I (K1) kinetics in these species.

Ginsenoside Re suppresses electromechanical alternans in cat and human cardiomyocytes

Ginseng botanicals are increasingly used as complementary or alternative medicines for a variety of cardiovascular diseases, yet little is known about their cellular actions in cardiac muscle. Electromechanical alternans (EMA) is a proarrhythmic cardiac abnormality that results from disturbances of intracellular Ca(2+) homeostasis. This study sought to determine whether a purified ginsenoside extract of ginseng, Re, exerts effects to suppress EMA and to gain insight into its mechanism of action. Alternans was induced by electrically pacing cardiomyocytes at room temperature. Re (> or = 10 nM) reversibly suppressed EMA recorded from cat ventricular and atrial myocytes and Langendorff-perfused cat hearts. In cat ventricular myocytes, Re reversibly suppressed intracellular Ca(2+) concentration ([Ca(2+)](i)) transient alternans. Re exerted no significant effects on baseline action potential configuration or sarcolemmal L-type Ca(2+) current (I(Ca,L)), Na(+) current, or total K(+) conductance. In human atrial myocytes, Re suppressed mechanical alternans and exerted no effect on I(Ca,L). In cat ventricular myocytes, Re increased [Ca(2+)](i) transient amplitude and decreased sarcoplasmic reticulum (SR) Ca(2+) content, resulting in an increase in fractional SR Ca(2+) release. In SR microsomes isolated from cat ventricles, Re had no effect on SR Ca(2+) uptake. Re increased the open probability of ryanodine receptors (RyRs), i.e., SR Ca(2+)-release channels, isolated from cat ventricles and incorporated into planar lipid bilayers. We concluded that ginsenoside Re suppresses EMA in cat atrial and ventricular myocytes, cat ventricular muscle, and human atrial myocytes. The effects of Re are not mediated via actions on sarcolemmal ion channels or action potential configuration. Re acts via a subcellular mechanism to enhance the opening of RyRs and thereby overcome the impaired SR Ca(2+) release underlying EMA.

Na+/Ca2+ exchanger inhibition exerts a positive inotropic effect in the rat heart, but fails to influence the contractility of the rabbit heart

The Na(+)/Ca(2+) exchanger (NCX) may play a key role in myocardial contractility. The operation of the NCX is affected by the action potential (AP) configuration and the intracellular Na(+) concentration. This study examined the effect of selective NCX inhibition by 0.1, 0.3 and 1.0 microM SEA0400 on the myocardial contractility in the setting of different AP configurations and different intracellular Na(+) concentrations in rabbit and rat hearts. The concentration-dependent effects of SEA0400 on I(Na/Ca) were studied in rat and rabbit ventricular cardiomyocytes using a patch clamp technique. Starling curves were constructed for isolated, Langendorff-perfused rat and rabbit hearts. The cardiac sarcolemmal NCX protein densities of both species were compared by immunohistochemistry. SEA0400 inhibited I(Na/Ca) with similar efficacy in the two species; there was no difference between the inhibitions of the forward or reverse mode of the NCX in either species. SEA0400 increased the systolic and the developed pressure in the rat heart in a concentration-dependent manner, for example, 1.0 microM SEA0400 increased the maximum systolic pressures by 12% relative to the control, whereas it failed to alter the contractility in the rabbit heart. No interspecies difference was found in the cardiac sarcolemmal NCX protein densities. NCX inhibition exerted a positive inotropic effect in the rat heart, but it did not influence the contractility of the rabbit heart. This implies that the AP configuration and the intracellular Na(+) concentration may play an important role in the contractility response to NCX inhibition.

Effects of Ropivacaine on Action Potential Configuration and Ion Currents in Isolated Canine Ventricular Cardiomyocytes

Despite the widespread clinical application of ropivacaine, there is little information on the cellular cardiac effects of the drug. In the current study, therefore, the concentration-dependent effects of ropivacaine on action potential morphology and the underlying ion currents were studied and compared with those of bupivacaine in isolated canine ventricular cardiomyocytes. Action potentials were recorded from the enzymatically dispersed cells using sharp microelectrodes. Conventional patch clamp and action potential voltage clamp arrangements were used to study the effects of ropivacaine on transmembrane ion currents. Ropivacaine induced concentration- and frequency-dependent changes in action potential configuration, including shortening of the action potentials, reduction of their amplitude and maximum velocity of depolarization, suppression of early repolarization, and depression of plateau. Reduction in maximum velocity of depolarization was characterized with an EC50 value of 81 +/- 7 microm at 1 Hz. Qualitatively similar results were obtained with bupivacaine (EC50 = 47 +/- 3 microm). Under voltage clamp conditions, a variety of ion currents were blocked by ropivacaine: L-type calcium current (EC50 = 263 +/- 67 microm), transient outward current (EC50 = 384 +/- 75 microm), inward rectifier potassium current (EC50 = 372 +/- 35 microm), rapid delayed rectifier potassium current (EC50 = 303 +/- 47 microm), and slow delayed rectifier potassium current (EC50 = 106 +/- 18 microm). Ropivacaine, similarly to bupivacaine, can modify cardiac action potentials and the underlying ion currents at concentrations higher than the usual therapeutic range. However, in cases of overdose, cardiac complications may be anticipated both during and after anesthesia due to the blockade of various ion currents.

Effects of rosiglitazone on altered electrical left ventricular papillary muscle activities of diabetic rat

The action potential configuration of the left ventricular papillary muscle as well as the rosiglitazone-dependent changes in ventricular papillary muscle action potential amplitude were studied, and the duration was studied and compared in both healthy and diabetic rats. In this study, we used four groups: (1) nondiabetic control animals (C), (2) rosiglitazone-treated nondiabetic control animals (C+RSG), (3) diabetic animals (D), and (4) rosiglitazone-treated diabetic animals (D+RSG). Diabetes was induced by a single intravenous (i.v.) injection of streptozotocin (STZ). Conventional microelectrode techniques were applied to record action potentials after the establishment of diabetes (8 weeks after STZ treatment). Resting membrane potential (RMP) was decreased significantly in both RSG-treated C and D rats (from -70.2 +/- 0.7 to -63.2 +/- 0.7 and from -69.2 +/- 0.4 to -61.2 +/- 0.4). C+RSG and D+RSG groups showed increase in action potential amplitude compared with C and D groups (from 67.1 +/- 0.8 to 68.2 +/- 0.5 and from 67.1 +/- 0.8 to 80.1 +/- 0.8 and from 68.2 +/- 0.5 to 79.3 +/- 0.3) Depolarization time was significantly prolonged in diabetic rats (12.1 +/- 0.4 to 27.5 +/- 0.9). However, this prolongation in D+RSG group was significantly lower according to D group (from 27.5 +/- 0.9 to 19.2 +/- 0.7). There was no difference between C and C+RSG rats (12.1 +/- 0.4 to 11.6 +/- 0.2). Half repolarization time was also prolonged in diabetic rats (17.5 +/- 0.6 to 59.9 +/- 1.0). Moreover, D+RSG rats showed a slight and statistically insignificant difference according D rats (59.9 +/- 1.0 to 55.9 +/- 1.7). C+RSG rats showed a slight significant increase in half repolarization time compared with C group (17.5 +/- 0.6 to 29.4 +/- 0.7). Treatment of rats with RSG markedly decreased insulin resistance and also increased insulin sensitivity of the heart. Our data suggest that the beneficial effects of RSG treatment on the electrical activities of the diabetic rat papillary appear to be due to the diminished K+ currents, partially related to the decrease of hyperglycemia.