ATP-sensitive Potassium Current Research Articles

Diadenosine-5-phosphate exerts A1-receptor-mediated proarrhythmic effects in rabbit atrial myocardium.

(1) Diadenosine polyphosphates have been described to be present in the myocardium and exert purinergic- and nonreceptor-mediated effects. Since the electrophysiological properties of atrial myocardium are effectively regulated by A(1) receptors, we investigated the effect of diadenosine pentaphosphate (Ap(5)A) in rabbit myocardium. (2) Parameters of supraventricular electrophysiology and atrial vulnerability were measured in Langendorff-perfused rabbit hearts. Muscarinic potassium current (I(K(ACh/Ado))) and ATP-sensitive potassium current (I(K(ATP))) were measured by using the whole-cell voltage clamp method. (3) Ap(5)A prolonged the cycle length of spontaneously beating Langendorff perfused hearts from 225+/-14 (control) to 1823+/-400 ms (Ap(5)A 50 micro M; n=6; P<0.05). This effect was paralleled by higher degree of atrio-ventricular block. Atrial effective refractory period (AERP) in control hearts was 84+/-14 ms (n=6). Ap(5)A>/=1 micro M reduced AERP (100 micro M, 58+/-11 ms; n=6). (4) Extrastimuli delivered to hearts perfused with Ap(5)A- or adenosine (>/= micro M)-induced atrial fibrillation, the incidence of which correlated to the concentration added to the perfusate. The selective A(1)-receptor antagonist CPX (20 micro M) inhibited the Ap(5)A- and adenosine-induced decrease of AERP. Atrial fibrillation was no longer observed in the presence of CPX. (5) The described Ap(5)A-induced effects in the multicellular preparation were enhanced by dipyridamole (10 micro M), which is a cellular adenosine uptake inhibitor. Dipyridamole-induced enhancement was inhibited by CPX. (6) Ap(5)A (</=1 mM) did neither induce I(K(Ado)) nor I(K(ATP)). No effect on activated I(K(Ado/ATP)) was observed in myocytes superfused with Ap(5)A. However, effluents from Langendorff hearts perfused with Ap(5)A 100 micro M activated I(K(Ado)) by using A(1) receptors. (7) Ap(5)A did not activate A(1) receptors in rabbit atrial myocytes. The Ap(5)A induced A(1)-receptor-mediated effects on supraventricular electrophysiology and vulnerability suggest that in the multicellular preparation Ap(5)A is hydrolyzed to yield adenosine, which acts via A(1) receptors. An influence on atrial electrophysiology or a facilitation of atrial fibrillation under conditions resulting in increased interstitial Ap(5)A concentrations might be of physiological/pathophysiological relevance.

Effect of acute hypoxia on ATP-sensitive potassium currents in substantia gelatinosa neurons of juvenile rats.

Although hypoxia is known to affect membrane excitability of various neurons by various mechanisms, the effects of hypoxia on substantia gelatinosa (SG) neurons have not yet been elucidated. In whole-cell or perforated patch-clamp recordings from SG neurons, we showed that acute hypoxia induces a reversible hyperpolarization of -6.1+/-1.3 mV of the resting membrane potential and an outwards current of 9.48+/-1.71 pA at a holding potential of -60 mV. The reversal potentials of the hypoxia-induced current depended on [K(+)](o). The hypoxia-induced hyperpolarization and outwards current were abolished completely by BaCl(2), but not by CsCl. Glibenclamide, a blocker of K(ATP) channels, blocked the hypoxia-induced hyperpolarization. Pretreatment with cromakalim, an opener of K(ATP) channels, occluded the hypoxia-induced hyperpolarization. Any alteration by hypoxia was not observed in the presence of an internal solution with a high [ATP] (10 mM). The above results suggest that hypoxia-induced hyperpolarization in SG neurons is mediated by activation of K(ATP) channels.

ATP-sensitive potassium channels in rat primary afferent neurons: the effect of neuropathic injury and gabapentin

ATP-sensitive potassium (K ATP) currents were examined in dorsal root ganglion neurons from neuropathic and control rats using whole-cell voltage clamp recordings. K ATP channel openers (diazoxide and pinacidil) enhanced, and the blocker glibenclamide inhibited an outward current in control neurons in a manner dependent on the pipette ATP concentration. Analysis of reversal potentials showed that this current is carried by K + ions. Outward current in cells from rats with peripheral nerve injury was not sensitive to modulators of K ATP channels. Gabapentin, a putative K ATP channel opener, had minimal effect on currents in either group of neurons. We conclude that normal primary afferent neurons express K ATP channels that conduct current which is eliminated by peripheral nerve injury. Gabapentin does not affect this current significantly.

The Origin of Calcium Overload in Rat Cardiac Myocytes Following Metabolic Inhibition With 2,4-Dinitrophenol

D. Hudman, R. D. Rainbow, C. L. Lawrence and N. B. Standen. The Origin of Calcium Overload in Rat Cardiac Myocytes Following Metabolic Inhibition With 2,4-Dinitrophenol. Journal of Molecular and Cellular Cardiology (2002) 34, 859–871. We have investigated the characteristics of the rise in cytoplasmic calcium that occurs when rat isolated cardiac ventricular myocytes are exposed to 2,4-dinitrophenol using conventional and confocal fluorescence microscopy and patch clamp. 2,4-dinitrophenol (200μM) caused cytoplasmic calcium to increase in two phases: (1) an initial rise in fluo-3 fluorescence of 36±2% that was maintained until rigor contraction; (2) a further progressive rise so that fluo-3 fluorescence had increased by 177±12% 535s after 2,4-dinitrophenol addition. Both phases were unaffected by removal of external Ca2+. 2,4-dinitrophenol caused mitochondrial depolarization, measured using tetramethyl rhodamine ethyl ester fluorescence. Mitochondrial depolarization was associated with a decrease in intra-mitochondrial calcium measured with rhod-2, and experiments on myocytes loaded with both fluo-3 and rhod-2 showed that fluo-3 fluorescence increased as rhod-2 fluorescence fell. The correlation of the onset of the second phase of the increase in cytoplasmic calcium with rigor suggested that this phase was consequent on ATP depletion. DNP also caused activation of an ATP-sensitive potassium current. Depletion of sarcoplasmic reticulum calcium stores by pretreatment with ryanodine, thapsigargin and caffeine prior to the addition of 2,4-dinitrophenol did not affect the initial increase in cytoplasmic calcium, but abolished phase 2. Our results suggest that the initial rise in cytoplasmic calcium seen on application of 2,4-dinitrophenol results from release of mitochondrial calcium because of mitochondrial depolarization, while the second phase is caused by progressive release of calcium from the sarcoplasmic reticulum following depletion of intracellular ATP.

Ionic Current Basis of Electrocardiographic Waveforms

Body surface electrocardiograms and electrograms recorded from the surfaces of the heart are the basis for diagnosis and treatment of cardiac electrophysiological disorders and arrhythmias. Given recent advances in understanding the molecular mechanisms of arrhythmia, it is important to relate these electrocardiographic waveforms to cellular electrophysiological processes. This modeling study establishes the following principles: (1) voltage gradients created by heterogeneities of the slow-delayed rectifier (I(Ks)) and transient outward (I(to)) potassium current inscribe the T wave and J wave, respectively; T-wave polarity and width are strongly influenced by the degree of intercellular coupling through gap-junctions. (2) Changes in [K+]o modulate the T wave through their effect on the rapid-delayed rectifier, I(Kr). (3) Alterations of I(Ks), I(Kr), and I(Na) (fast sodium current) in long-QT syndrome (LQT1, LQT2, and LQT3, respectively) are reflected in characteristic QT-interval and T-wave changes; LQT1 prolongs QT without widening the T wave. (4) Accelerated inactivation of I(Na) on the background of large epicardial I(to) results in ST elevation (Brugada phenotype) that reflects the degree of severity. (5) Activation of the ATP-sensitive potassium current, I(K(ATP)), is sufficient to cause ST elevation during acute ischemia. These principles provide a mechanistic cellular basis for interpretation of electrocardiographic waveforms.

Uncoupling of ATP-sensitive potassium current from cell metabolism due to antisense oligonucleotides against sulfonylurea receptor in guinea-pig atrial myocytes.

ATP-sensitive K+ current (IK(ATP)) plays an important role in the regulation of cardiac electrical activity. In the myocardium, IK(ATP) is regulated by the sulfonylurea receptor (SURIIA) which mediates the inhibition of IK(ATP) due to glibenclamide (Gli). The role played by SURIIA in the sensitivity of IK(ATP) to metabolic inhibition is unclear. We studied the effect of SURIIA antisense oligonucleotides (ODNs) on the properties of IK(ATP) in cultured guinea-pig atrial myocytes. IK(ATP) was measured by the whole-cell voltage clamp method and was activated with cromakalim (Cro; 200 microns) and dinitrophenole (DNP; 100 microns). Mean IK(ATP) density activated by DNP and Cro in nonincubated cells was 117 +/- 12 pA/pF (n = 17) and 17 +/- 9 pA/pF (n = 16) respectively. No significant difference was observed after incubation with nODN [DNP: 121 +/- 13 pA/pF (n = 20); Cro:19 +/- 4 pA/pF (n = 8)]. Cells incubated with ODNs showed a significant reduction of IK(ATP) due to DNP (19 +/- 13 pA/pF; P < 0.05, n = 6), whereas Cro-induced IK(ATP) was unaffected (16 +/- 8 pA/pF, n = 8). The effectiveness of DNP-induced metabolic inhibition was apparent in a concomitant reduction of the nucleotide-phosphate dependent muscarinic K+ current (inhibition of IK(ACh) in ODN incubated myocytes without activation of IK(ATP)). The ATP sensitivity of IK(ATP) appears mediated by SURIIA. Activation of this current by Cro seems to be SURIIA-independent. ODN-induced metabolic uncoupling of IK(ATP) may be a useful experimental tool. A reduced sensitivity of IK(ATP) to intracellular ATP concentrations may be of clinical interest.

Effect of GLG-V-13, a class III antiarrhythmic agent, on potassium currents in rabbit ventricular myocytes

The effects of a new Class III antiarrhythmic drug, GLG-V-13, on the 4-aminopyridine sensitive transient outward current, on the inward rectifier potassium current, on the ATP sensitive potassium current and on the rapid and slow components of the delayed rectifier potassium current were studied in single rabbit ventricular myocytes using the whole-cell voltage-clamp technique. GLG-V-13 blocked the rapid component of the delayed rectifier potassium current in a dose-dependent manner, with an estimated EC 50 value of 0.36 μM. At high concentration, the slow component of the delayed rectifier potassium current was also depressed by the drug (40% effect at 10 μM concentration). The transient outward current, the inward rectifier potassium current and the ATP sensitive potassium current were not influenced by GLG-V-13, even at 10 μM concentration. Thus, GLG-V-13 blocks predominantly the rapid component of the delayed rectifier potassium current which may play a significant role in the prolongation of repolarization by the drug in ventricular tissue.

Propafenone blocks ATP-sensitive K + channels in rabbit atrial and ventricular cardiomyocytes

Propafenone, a class I antiarrhythmic agent, inhibits several membrane currents ( I Na, I Ca, I K, I to), however, its effects on ATP-sensitive potassium current ( I K ATP ) of cardiac cells have not been tested. We evaluated the blocking effects of 0.1 to 100 μM propafenone applications at 35°C on the whole-cell I K ATP as triggered by dinitrophenol (75 μM) in adult rabbit dissociated atrial and ventricular cardiomyocytes in comparison. The block of I K ATP by propafenone was dose-dependent, fully reversible and voltage-independent. The dose–response relation, as evaluated at 0 mV for atrial myocytes (ED 50=1.26±0.17 μM, Hill number=1.25±0.22) was significantly shifted to the left vs. that in ventricular myocytes (ED 50=4.94±0.59 μM, Hill number=1.22±0.14). It is concluded that propafenone blocks cardiac I K ATP at a single site with 4 times higher affinity for the drug in atrial myocytes. This block of cardiac I K ATP might play a role in the beneficial and adverse effects of the drug.

Evidence for an early G1 ionic event necessary for cell cycle progression and survival in the MCF-7 human breast carcinoma cell line.

The mechanism of the G0/G1 arrest and inhibition of proliferation by quinidine, a potassium channel blocker, was investigated in a tissue culture cell line, MCF-7, derived from a human breast carcinoma. The earliest measurable effect of quinidine on the cell cycle was a decrease in the fraction of cells in S phase at 12 hr, followed by the accumulation of cells in G1/G0 phases at 30 hr. Arrest and release of the cell cycle established quinidine as a cell synchronization agent, with a site of arrest in early G1 preceding the lovastatin G1 arrest site by 5-6 hr. There was a close correspondence among the concentration-dependent arrest by quinidine in G1, depolarization of the membrane potential, and the inhibition of ATP-sensitive potassium currents, supporting a model in which hyperpolarization of the membrane potential and progression through G1 are functionally linked. Furthermore, the G1 arrest by quinidine was overcome by valinomycin, a potassium ionophore that hyperpolarized the membrane potential in the presence of quinidine. With sustained exposure of MCF-7 cells to quinidine, expression of the Ki67 antigen, a marker for cells in cycle, decreased, and apoptotic and necrotic cell death ensued. We conclude that MCF-7 cells that fail to progress through the quinidine-arrest site in G1 die.

Electrophysiological and metabolic characterization of single beta-cells and islets from diabetic GK rats.

We have used the whole-cell recording technique to determine whether ATP-sensitive potassium (K[ATP]) currents, voltage-dependent Ca2+ currents, and exocytosis are different in single beta-cells from pancreatic islets of Goto-Kakizaki (GK) rats, a novel model of NIDDM, and normal rats. In addition, we have also measured the insulin secretory responses, ATP content, and the rate of glucose metabolism in intact islets. Although the glucose sensitivity of the K(ATP) current was similar between GK rats and controls, in the absence of glucose, K(ATP) current density was larger in GK rats, which resulted in a more hyperpolarized membrane potential. Whole-cell Ca2+ currents were similar. By monitoring the cell capacitance with a fixed intracellular solution, no difference was detected in the exocytotic responses of beta-cells from normal and GK rats. In islets from GK rats, the rates of glucose utilization ([3H]H2O production from 5-[3H]glucose) and oxidation ([14C]CO2 production from U-[14C]glucose) were not significantly different from controls. Insulin secretion, however, was impaired (by 50%), and this was paralleled by a smaller increase in ATP content in response to stimulation by 10 mmol/l glucose in islets from GK rats when compared with controls. Under conditions in which K(ATP) channels were held open and the effects of glucose were independent of membrane potential, insulin release was still significantly lower in GK rat islets than in controls. These findings suggest that the impaired insulin secretion in islets from GK rats does not simply result from a failure to close K(ATP) channels, nor does it result from an impairment in calcium secretion coupling.

Priming effect of adenosine on K(ATP) currents in intact ventricular myocytes: implications for preconditioning.

Activation of protein kinase C (PKC) by the phorbol ester phorbol 12-myristate 13-acetate (PMA) has been shown to shorten the time to turn on ATP-sensitive potassium currents (I(K,ATP)) during metabolic inhibition (MI) but only when adenosine (Ado) is included. In the present study we tested whether pretreatment with Ado could mimic the effect of PMA in isolated rabbit ventricular myocytes. When I(K,ATP) was measured by conventional whole cell clamp, pretreatment with 100 microM Ado did not alter the time to I(K,ATP) activation: 13.5 +/- 2.1 vs. 12.4 +/- 1.9 min with Ado during MI. We repeated the experiment using the perforated patch technique. Consistent with the previous results in conventional whole cell patch recordings, the time to turn on I(K,ATP) during MI (with Ado included) was shortened from 27.1 +/- 2.2 to 12.6 +/- 2.4 min (P < 0.01) when cells were pretreated with PMA and Ado was included during MI. In contrast to conventional whole cell recordings, Ado pretreatment also abbreviated the time for I(K,ATP) activation during MI (with Ado included) to 16.4 +/- 1.8 min. This effect was partially eliminated by simultaneous administration of an Ado receptor blocker or a PKC inhibitor during Ado pretreatment, suggesting that pretreatment with Ado stimulates PKC by activating Ado receptors. Our results demonstrate that Ado can prime I(K,ATP) during subsequent MI in the presence of Ado. This priming effect appears to be mediated by PKC upregulation of the channel. These results support the notion that Ado plays a dual role to initiate and to mediate ischemic preconditioning and links the roles of Ado receptors, PKC, and I(K,ATP) in ischemic preconditioning.

Electrical remodeling in atrial fibrillation. Time course and mechanisms.

Atrial fibrillation is self-perpetuating, suggesting that the tachyarrhythmia causes electrophysiological changes that contribute to the progressive nature of the disease. In animal models, pacing-induced rapid atrial rates result in sustained atrial fibrillation. This is mediated by shortening of refractory periods termed electrical remodeling. The purpose of the present study was to characterize the time course of electrical remodeling and to define mechanisms of the phenomenon. Closed-chest dogs were anesthetized, pretreated with atropine and propranolol, and subjected to 7 hours of atrial pacing at 800 bpm. The effective and absolute refractory periods (ARP and ERP) were measured during and after rapid pacing, and transvenous endocardial biopsy specimens were examined using electron microscopy. Despite autonomic blockade and the absence of change in right atrial pressure, persistent atrial tachycardia caused ARP and ERP to fall by > 10%. Electrical remodeling developed quickly, with more than half of the phenomenon occurring during the first 30 minutes of high-rate pacing. Pretreatment with glibenclamide in doses sufficient to block the ATP-sensitive potassium current had no effect. Atrial electrical remodeling was blocked by verapamil and accentuated by hypercalcemia. Biopsy specimens from controls subjected to rapid pacing showed mitochondrial swelling consistent with calcium overload. Biopsies from verapamil-treated animals were normal. Atrial electrical remodeling develops quickly, is progressive, and may be persistent. Shifts in autonomic tone, atrial stretch, or depletion of high-energy phosphates do not contribute significantly to the phenomenon. Results of the study suggest that atrial electrical remodeling is mediated by rate-induced intracellular calcium overload.

Mutation of the pancreatic islet inward rectifier Kir6.2 also leads to familial persistent hyperinsulinemic hypoglycemia of infancy.

Closure of ATP-sensitive potassium channels in pancreatic islet beta-cells initiates a cascade of events that leads to insulin secretion. beta-Cell ATP-sensitive potassium currents can be reconstituted by coexpression of the inward rectifier Kir6.2 and the sulfonylurea receptor (SUR), a member of the ATP-binding cassette superfamily. Mutations in SUR have been identified in individuals affected with familial persistent hyper-insulinemic hypoglycemia of infancy (PHHI), an autosomal recessive disorder of glucose metabolism which is linked to chromosome 11p15.1 and characterized by unregulated secretion of insulin and profound hypoglycemia. Because the Kir6.2 locus is within 5 kilobases (kb) of the SUR gene on chromosome 11p15.1 and it is a necessary member of the beta-cell KATP channel, we considered Kir6.2 as a candidate gene for PHHL we identified a homozygous point mutation in Kir6.2 in the genomic DNA of a child, severely affected with PHHI, from a consanguineous family. This mutation is predicted to disrupt the conserved alpha-helical second transmembrane (M2) domain of the inward rectifier by substitution of a proline for a leucine residue (L147P). Mutation of Kir6.2, like SUR, appears to lead to the PHHI phenotype suggesting that Kir6.2 is necessary, although not sufficient, for normal regulation of insulin release.

Electrophysiologic effects of potassium channel openers

Potassium-channel openers or activators have been introduced as a new class of antihypertensive and antianginal agents that act by increasing membrane conductance to potassium, mainly through augmentation of the ATP-sensitive potassium current. Recent in vitro studies have shown that K(+)-channel openers exert concentration-dependent effects on cardiac electrophysiology. A shortening of the cardiac action potential by acceleration of repolarization has been reported in multicellular preparations as well as in isolated myocytes. However, drug concentrations that affect the action potential duration of myocardial cells are considerably higher (10- to 100-fold) than those needed for effects on vascular smooth muscle cells. Studies in which mostly high concentrations of K(+)-channel openers were used have demonstrated that these drugs may accelerate automaticity and may promote reentrant activity. Particular interest has focused on the question whether opening of potassium channels may be potentially arrhythmogenic in the setting of acute myocardial ischemia. On the other hand, recent studies have shown that K(+)-channel openers are effective in suppressing polymorphic ventricular tachyarrhythmias induced by early afterdepolarizations and triggered activity in vivo. The clinical relevance of these experimental studies to the clinical situation is still unclear. Some K(+)-channel openers have been shown to produce electrocardiographic T-wave changes in patients in whom their effectiveness as antihypertensives was tested. However, this effect was not associated with adverse effects and has not been demonstrated for all compounds.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of glibenclamide, forskolin, and isoprenaline on the parallel activation of KATP and reduction of IK by cromakalim in cardiac myocytes.

The aim was to investigate the effect of activation of ATP sensitive potassium channels by cromakalim on the delayed rectifier potassium current (IK) in guinea pig ventricular myocytes. Experiments were carried out in the absence and presence of forskolin or isoprenaline to promote phosphorylation of IK, and in the presence of glibenclamide to block the ATP sensitive potassium current (IK(ATP)). Single cells were isolated from guinea pig ventricle. Potassium currents were studied under voltage clamp conditions. The delayed rectifier was measured as an outward tail current upon repolarisation to a holding potential of -40 mV following depolarising steps to +40 mV. Induction of IK(ATP) was indicated by changes in the holding current. Exposure to 20 microM cromakalim caused a significant increase of 244(SEM 47)% in the holding current and simultaneous decreases of 22(5)% in the rapid component (IKr) and 45(5)% in the slow component (IKs) of IK. Exposure of the cells to 5 microM forskolin or 100 nM isoprenaline reduced both these effects of cromakalim: with forskolin, the holding current increased by 59(17)%, IKr was reduced by 9(3)%, and IKs by 23(3)%; with isoprenaline, the holding current increased by 100(36)%, IKr was not significantly changed, and IKs was reduced by 27(5)%. With 10 microM glibenclamide present, the only significant effect of cromakalim was reduction of IKs [by 11(3)%]. Cromakalim caused decreases in both components of IK which developed in parallel with activation of IK(ATP). The observations that forskolin, isoprenaline, or glibenclamide all reduced the effects of cromakalim on both IK and IK(ATP) may result from separate effects on the two channel pathways, but are also consistent with the single hypothesis that cromakalim induces an interconversion of potassium channels which is reduced when potassium channels are modified by these three drugs.

Blockers of ATP sensitive potassium current are of potential benefit in ischaemic heart disease.

Blockers of ATP sensitive potassium current are of potential benefit in ischaemic heart disease.

Enhancement of ATP‐sensitive potassium current in cat ventricular myocytes by beta‐adrenoreceptor stimulation.

1. To address the questions of whether beta-adrenoreceptor stimulation can augment ATP-sensitive potassium current (IK(ATP)), and what the mechanism of such an effect might be, action potentials and whole-cell ionic currents were recorded from adult cat cardiac ventricular myocytes using a conventional whole-cell patch technique. 2. An outwardly directed, ohmic, non-inactivating, glyburide (10 microM)-sensitive current reversing near the reversal potential for potassium (EK) developed slowly (10-25 min) in cells dialysed with an ATP-free pipette (intracellular) solution. During this time, action potential duration markedly decreased while the resting membrane potential hyperpolarized closer to EK. Extended (> 30 min) periods of internal dialysis with ATP-free solution eventually resulted in run-down of the outward current. 3. Externally applied isoprenaline (1 microM) caused a rapidly developing (< or = 60 s), sustained enhancement of a glyburide (10 microM)-sensitive IK(ATP) in cells internally dialysed with ATP-free solution. IK(ATP) remained elevated even after the isoprenaline was removed, and subsequent applications of the beta-agonist failed to increase IK(ATP) further. Half-maximal isoprenaline stimulation of IK(ATP) occurred at a concentration of approximate of 1.5 nM. 4. Pretreatment with propranolol (1 microM) prevented the enhancement of IK(ATP) by a beta-agonist. 5. Isoprenaline-induced IK(ATP) could be blocked by either internal application of GDP-beta-S (2-5 mM) or pretreatment with cholera toxin (1-10 microgram ml-1, > 18 h). Pretreatment with pertussis toxin (1-2 microgram ml-1, > 18 h) did not attenuate the isoprenaline response, whereas internally applied GTP-gamma-S (100 microM) or F- (20 mM) caused IK(ATP) to increase rapidly in the absence of the beta-agonist. 6. Although externally applied forskolin (10 microM) also stimulated IK(ATP), neither 1,9-dideoxyforskolin (10 microM) nor 8-(4-chlorophenylthio)-cAMP (200 microM) had any effect on the current. Internal application of the adenylate cyclase inhibitor 2'-deoxyadenosine-3'-monophosphate (100 microM) resulted in a reduction in the response to isoprenaline, while internal application of a protein kinase A inhibitor (PKI5-24, 22.5 microM) did not attenuate the response to the beta-agonist. 7. IK(ATP) developed slowly during internal dialysis with ATP-free solution.(ABSTRACT TRUNCATED AT 400 WORDS)

Blockade of 2,4-dinitrophenol induced ATP sensitive potassium current in guinea pig ventricular myocytes by class I antiarrhythmic drugs.

The aim was to assess the effects of various antiarrhythmic drugs on 2,4-dinitrophenol (DNP) induced outward current (IDNP), presumably the ATP sensitive K+ current (IK,ATP) of isolated cardiac cells and to discuss mechanisms involved in the hypoglycaemia which occurs in patients on these drugs. The quasi-steady state current-voltage relationship from the isolated guinea pig ventricular cells was measured using whole cell voltage clamp techniques with a ramp pulse programme. The effects of seven different antiarrhythmic drugs on IDNP were examined. Action potentials were elicited at a rate of 0.2 Hz by an intracellular current injection. DNP (50 mumol.litre-1) increased the quasi-steady state outward current at potentials positive to about -60 mV. This current (IDNP) was completely inhibited by the subsequent application of glibenclamide (1 mumol.litre-1), thereby suggesting that the IDNP is probably IK,ATP. Cibenzoline (10 mumol.litre-1, class Ia), disopyramide (30 mumol.litre-1, class Ia), and procainamide (100 mumol.litre-1, class Ia) significantly inhibited the IDNP by 95.5(SD 11.3)%, 77.8(21.2)%, and 76.4(23.9)% respectively. Flecainide (class 1c) inhibited the IDNP by 66.9(23.9)% at 10 mumol.litre-1 but not at 2 mumol.litre-1. Mexiletine (30 mumol.litre-1, class Ib), pilsicainide (50 mumol.litre-1, class Ic), and E4031 (10 mumol.litre-1, class III) at concentrations as high as approximately fivefold the clinically effective blood levels, did not suppress IDNP. Except for 10 mumol.litre-1 flecainide, all the concentrations listed above which blocked IDNP were within twofold of the clinical blood concentrations documented to be effective for suppression of arrhythmias. Cibenzoline, disopyramide, and procainamide, but not flecainide, belong to class Ia antiarrhythmic drugs. All these class Ia antiarrhythmic drugs "shortened" the action potential duration of guinea pig ventricular cells, an opposite change to that noted for multicellular preparations, eg, guinea pig papillary muscles. Class Ia antiarrhythmic drugs (cibenzoline, disopyramide, and procainamide) inhibit IDNP (presumably IK,ATP) in guinea pig ventricular cells within a range of therapeutic concentrations. This inhibitory effect of IK,ATP can probably explain the hypoglycaemia which occurs in some patients receiving these drugs, and the prolongation of the action potential duration alleged to occur in "superfused" papillary muscles.

ATP-sensitive potassium channels in neonatal and adult rabbit ventricular myocytes.

The properties of the ATP-sensitive potassium (KATP) current were studied in freshly isolated rabbit ventricular myocytes using the patch clamp technique. Removing ATP from the bath (intracellular) solution activated a large K+ conductance in patches from neonatal cells with properties similar to those of KATP channels in other preparations. In membrane patches from neonatal ventricular myocytes, the density of KATP channels was higher than the density of inwardly rectifying K+ channels and the mean patch KATP current was approximately 10 times that of the inwardly rectifying K+ current, at a patch membrane potential of -60 mV. Glibenclamide (10 microM) in the bath solution decreased the number of functional KATP channels, the open-state probability, and the mean patch membrane current. The single-channel conductance of the KATP channel was dependent on the external K+ concentration, and the relationship between channel conductance and external K+ concentration was fit by an exponential equation. In addition, the voltage dependence, channel density, and open-state probability of this channel were compared between neonatal and adult isolated ventricular myocytes. The single-channel conductance and channel density of the KATP channel in neonatal myocytes were significantly smaller than in adult cells. These results suggest that age-related changes occur in the properties of KATP channels.

ATP-sensitive potassium current in guinea pig ventricular myocytes is inhibited by extracellular pyruvate—is there a role for intracellular pyruvate?

ATP-sensitive potassium current in guinea pig ventricular myocytes is inhibited by extracellular pyruvate—is there a role for intracellular pyruvate?