Effects of metoprolol on action potential and membrane currents in guinea-pig ventricular myocytes

Summary The effects of the beta-adrenoceptor antagonist metoprolol on action potentials and membrane currents were studied in single guinea-pig ventricular myocytes. The experiments were carried out using the nystatin-method of whole-cell technique. This method was used in order to prevent the run-down of the calcium current. Metoprolol at concentrations of 10–100 mol/l shortened action potential in a dose-dependent way. The drug only decreased resting membrane potential at a concentration of 100 mol/1 in two out of five cells. Under voltage-clamp conditions, metoprolol blocked the high threshold calcium current at concentrations of 30 and 100 mol/l to 82 ± 4% and 73 ± 5% from control, respectively. The drug decreased the inward rectifying potassium current in a concentration-dependent manner. This effect was evident for inward current at voltages negative to the apparent reversal potential and for outward current at voltages between –30 and –80 mV. This blocking effect on the inward rectifying potassium current can explain the effect on resting membrane potential. At voltages positive to –30 mV metoprolol increased a time-independent outward current. This metoprolol-enhanced outward current was blocked by barium and cesium. This result suggests that the metoprolol-enhanced current is carried by potassium. The current component enhanced by metoprolol was not sensitive to glibenclamide and tetraethylammonium applied externally, which suggests that the adenosine triphosphate-sensitive channel is not the target of metoprolol. The activation of this time-independent outward current by metoprolol and the blocking effects on the calcium current seem to explain the shortening in action potential induced by the drug. Send offprint requests to J. Sánchez-Chapula at the above address     

Effects of propofol and enflurane on action potentials, membrane currents and contraction of guinea-pig isolated ventricular myocytes.

1. The effects of two general anaesthetics, propofol and enflurane, on electrical activity and contractions were investigated in single myocytes isolated from guinea-pig ventricles. 2. Propofol and enflurane depressed the plateau and shortened the duration of action potentials. 3. Under voltage-clamp conditions, propofol and enflurane reduced the amplitude of inward calcium current and of additional inward current activated by cytosolic calcium. 4. Contractions (measured with an optical technique) accompanying either action potentials or second inward currents (in response to depolarizations to 0 mV) were reduced by both anaesthetics. The mechanisms for calcium entry during contractions accompanying pulses to positive potentials such as +60 mV are thought to differ from those accompanying second inward currents which are evoked by pulses from -40 to 0 mV. Enflurane enhanced the amplitudes of contractions accompanying pulses to positive potentials; in contrast these contractions were depressed by propofol. 5. In experiments where recovery processes were investigated by use of pairs of voltage-clamp pulses with a variable interval between them, enflurane but not propofol slowed the recovery of contractions and calcium-activated 'tail' currents. These observations are consistent with the hypothesis that enflurane may impair calcium handling by the sarcoplasmic reticulum whereas propofol has little, if any, effect at this site. 6. In conclusion, the actions of propofol and enflurane on second inward currents contribute to their effects on action potentials and contraction. The negative inotropic effect of both anaesthetics may result partly from reduced calcium influx to trigger contraction, and for enflurane, partly from an impairment of calcium handling by the sarcoplasmic reticulum.     

Risperidone prolongs cardiac action potential through reduction of K+ currents in rabbit myocytes

Prolongation of QT interval by antipsychotic drugs is an unwanted side effect that may lead to ventricular arrhythmias. The antipsychotic agent risperidone has been shown to cause QT prolongation, especially in case of overdosage. We investigated risperidone effects on action potentials recorded from rabbit Purkinje fibers and ventricular myocardium and on potassium currents recorded from atrial and ventricular rabbit isolated myocytes. The results showed that (1) risperidone (0.1-3 microM) exerted potent lengthening effects on action potential duration in both tissues with higher potency in Purkinje fibers and caused the development of early afterdepolarizations at low stimulation rate; (2) risperidone (0.03-0.3 microM) reduced significantly the current density of the delayed rectifier current and at 30 microM decreased the transient outward and the inward rectifier currents. This study might explain QT prolongation observed in some patients treated with risperidone and gives enlightenment on the risk of cardiac adverse events.     

Effects of MS-551, a new class III antiarrhythmic drug,on action potential and membrane currents in rabbit ventricular myocytes.

1. Electrophysiological effects of MS-551, a new class III antiarrhythmic drug, were examined and compared with those of (+)-sotalol in rabbit ventricular cells. 2. In rabbit ventricular muscles stimulated at 1.0 Hz, MS-551 (0.1-10 microM) and (+)-sotalol (3-100 microM) prolonged action potential duration (APD) and effective refractory period without affecting the maximum upstroke velocity of phase 0 depolarization (Vmax). The class III effect of MS-551 was approximately 30 times more potent than that of (+)-sotalol. 3. Class III effects of MS-551 and (+)-sotalol showed reverse use-dependence, i.e., a greater prolongation of APD at a longer cycle length. 4. In rabbit isolated ventricular cells, 3 microM MS-551 and 100 microM sotalol inhibited the delayed rectifier potassium current (IK) which was activated at more positive potentials than -50 mV and saturated around +20 mV. 5. MS-551 at a higher concentration of 10 microM decreased the transient outward current (Ito) and the inward rectifier potassium current (IK1) although 100 microM sotalol failed to inhibit these currents. 6. MS-551 is a non-specific class III drug which can inhibit three voltage-gated K+ channels in rabbit ventricular cells.     

Effects of BDF 9198 on action potentials and ionic currents from guinea-pig isolated ventricular myocytes

BDF 9198 (a congener of DPI 201 - 106 and BDF 9148) was found to be a positive inotrope on guinea-pig isolated ventricular muscle strips. The effects of BDF 9198 on action potentials and ionic currents from guinea-pig isolated ventricular myocytes were studied using the whole cell patch clamp method. In normal external solution, at 37 degrees C, action potential duration at 50% repolarization (APD(50)) was 167.4+/-8.36 ms (n=37). BDF 9198 produced a concentration-dependent increase in APD(50) (no significant increase at 1x10(-10) M; and APD(50) values of 273.03+/-35.8 ms at 1x10(-9) M; n=6, P<0.01 and 694.7+/-86.3 ms at 1x10(-7) M; P<0.001, n=7). At higher concentrations in the range tested, BDF 9198 also induced early and delayed and after-depolarizations. Qualitative measurements of I(Na) with physiological [Na](o) showed prolongation of the current by BDF 9198, and the appearance of transient oscillatory inward currents at high concentrations. Quantitative recording conditions for I(Na) were established using low external [Na] and by making measurements at room temperature. The current - voltage relation, activation parameters and time-course of I(Na) were similar before and after a partial blocking dose of Tetrodotoxin (TTX, 1 microM), despite a 2 fold difference in current amplitude. This suggests that voltage-clamp during flow of I(Na) was adequately maintained under our conditions. Selective measurements of I(Na) at room temperature showed that BDF 9198 induced a concentration-dependent, sustained component of I(Na) (I(Late)) and caused a slight left-ward shift in the current - voltage relation for peak current. The drug-induced I(Late) showed a similar voltage dependence to peak current in the presence of BDF 9198. Both peak current and I(Late) were abolished by 30 microM TTX and were sensitive to external [Na]. Inactivation of control I(Na) during a 200 ms test pulse to -30 mV followed a bi-exponential time-course. In addition to inducing a sustained current component, BDF 9198 left the magnitude of the fast inactivation time-constant unchanged, but increased the magnitude of the slow inactivation time-constant. Additional experiments with a longer pulse (1 s) raised the possibility that in the presence of BDF 9198, I(Na) inactivation may be comprised of more than two phases. No significant effects of 1x10(-6) M BDF 9198 were observed on the L-type calcium current, or delayed and inward rectifying potassium currents measured at 37 degrees C. It is concluded that the prolongation of APD(50) by BDF 9198 resulted from selective modulation of I(Na). Reduced current inactivation induced a persistent I(Na), increasing the net depolarizing current during the action potential. This action of the drug indicates a potential for 'QT prolongation' of the ECG. The observation of after-depolarizations suggests a potential for proarrhythmia at some drug concentrations.     

Effects of halothane on membrane currents associated with contraction in single myocytes isolated from guinea-pig ventricle.

1. The effects of halothane on electrical activity and contraction were investigated in single myocytes isolated from guinea-pig ventricle. 2. Halothane depressed the plateau and shortened the duration of action potentials. 3. Halothane also reduced the amplitude of inward calcium currents and of additional inward current activated by cytosolic calcium under voltage-clamp conditions. 4. Contractions (measured by an optical technique) accompanying either action potentials or calcium currents were reduced by halothane. However, the extent of attenuation of contraction was greater than when a similar level of calcium channel blockade was induced by application of verapamil. 5. Actions of halothane on calcium-activated tail currents in double-pulse experiments were consistent with reduction by halothane of the cytosolic calcium transient, perhaps as a consequence of reduced uptake of calcium into sarcoplasmic reticulum stores. 6. It is concluded that the actions of halothane on inward currents contribute to its effects on action potentials. The reduction in contraction caused by halothane may result partly from a reduced influx of calcium to trigger contraction, and partly by a reduced release of calcium from sarcoplasmic reticulum stores.     

Comparative effects of clarithromycin on action potential and ionic currents from rabbit isolated atrial and ventricular myocytes

Prolongation of QT interval by several antibacterial drugs is an unwanted side effect that may be associated with development of ventricular arrhythmias. The macrolide antibacterial agent clarithromycin has been shown to cause QT prolongation. To determine the electrophysiologic basis for this arrhythmogenic potential, we investigated clarithromycin effects on (i). action potentials recorded from rabbit Purkinje fibers and atrial and ventricular myocardium using conventional microelectrodes and (ii). potassium and calcium currents recorded from rabbit atrial and ventricular isolated myocytes using whole-cell patch clamp recordings. We found that (i). clarithromycin (3-100 microM) exerted concentration-dependent lengthening effects on action potential duration in all tissues, with higher efficacy and reverse frequency-dependence in Purkinje fibers. However, clarithromycin did not cause development of early afterdepolarizations, and the parameters other than action potential duration were almost unaffected; (ii). clarithromycin (10-100 microM) reduced the delayed rectifier current. Significant blockade (approximately 30%) was found at the concentration of 30 microM. At 100 microM, it decreased significantly the maximum peak of the calcium current amplitude but failed to alter the transient outward and inwardly rectifier currents. It was concluded that these effects might be an explanation for the QT prolongation observed in some patients treated with clarithromycin.     

Effects of ropinirole on action potential characteristics and the underlying ion currents in canine ventricular myocytes

In spite of its widespread clinical application, there is little information on the cellular cardiac effects of the dopamine receptor agonist ropinirole. In the present study, therefore, the concentration-dependent effects of ropinirole on action potential morphology and the underlying ion currents were studied in enzymatically dispersed canine ventricular cardiomyocytes using standard microelectrode, conventional whole-cell patch clamp, and action potential voltage clamp techniques. At concentrations?≥?1 μM, ropinirole increased action potential duration (APD₉₀) and suppressed the rapid delayed rectifier K⁺ current (I _Kr) with an IC₅₀ value of 2.7?±?0.25 μM and Hill coefficient of 0.92?±?0.09. The block increased with increasing depolarizations to more positive voltages, but paradoxically, the activation of I _Kr was accelerated by 3 μM ropinirole (time constant decreased from 34?±?4 to 14?±?1 ms). No significant changes in the fast and slow deactivation time constants were observed with ropinirole. At higher concentrations, ropinirole decreased the amplitude of early repolarization (at concentrations?≥?10 μM), reduced the maximum rate of depolarization and caused depression of the plateau (at concentrations?≥?30 μM), and shortened APD measured at 50% repolarization (at 300 μM) indicating a concentration-dependent inhibition of I _to, I _Na, and I _Ca. Suppression of I _Kr, I _to, and I _Ca has been confirmed under conventional patch clamp and action potential voltage clamp conditions. I _Ks and I _K1 were not influenced significantly by ropinirole at concentrations less than 300 μM. All these effects of ropinirole were fully reversible upon washout. The results indicate that ropinirole treatment may carry proarrhythmic risk for patients with inherited or acquired long QT syndrome due to inhibition of I _Kr—especially in cases of accidental overdose or intoxication.     

Tonic and use-dependent block of sodium currents in isolated cardiac myocytes by bisaramil.

1. The effects of bisaramil on sodium currents in rat isolated cardiac myocytes were examined by use of tight-seal, whole-cell patch clamp techniques. Bisaramil produced a concentration-dependent, readily reversible reduction in peak transient sodium current. When the sodium current was evoked at 3 s intervals the estimated ED50 for bisaramil was about 11 microM. 2. Bisaramil (16 microM) produced a shift in the inactivation curve to hyperpolarized potentials of about 10 mV, but produced no change in the voltage-dependence of activation. 3. The block of the sodium current by bisaramil showed a profound use-dependence. A concentration of 10 microM produced a considerable block of the current with repeated stimulation. The recovery from block was biphasic, showing fast and slow components which had time constants of about 40 ms and 5 s respectively. 4. Bisaramil produced little tonic block of the sodium current at concentrations of 100 microM; at 300 microM it produced tonic block of around 50%, with extreme use-dependence. 5. Bisaramil appeared not to interact primarily with the inactivated form of the channel, since lengthening the depolarizing pulses did not affect the degree of block produced.     

Effects of N-acetylprocainamide and sotalol on ion currents in isolated guinea-pig ventricular myocytes.

The effects of N-acetylprocainamide (NAPA) and sotalol on membrane current systems of guinea-pig ventricular myocytes were examined and compared with those of quinidine using patch-clamp techniques. All of the drugs prolonged the action potential duration (i.e. Class III effect) in isolated guinea-pig papillary muscles. In isolated ventricular cells. NAPA (300 microM) and sotalol (100 microM) produced a decrease in the delayed outward potassium current (IK) concomitantly with a slight decrease in the calcium current (ICa), which was similar to quinidine (10 microM). NAPA also slightly depressed the inward rectifier potassium current (IKrect). Thus, NAPA and sotalol both inhibited IK, and this action appears to be mainly responsible for their Class III effect.     

Effects of bupivacaine on membrane currents of guinea-pig ventricular myocytes

The effects of bupivacaine on the membrane currents of single guinea-pig ventricular myocytes were investigated using the whole-cell patch-clamp technique. Bupivacaine decreased the inward calcium current in a concentration-dependent way at concentrations of 10 microM and higher. Bupivacaine also decreased the delayed outward current without modifying the inward-rectifying potassium current. From these results it can be concluded that bupivacaine, at concentrations lower than 10 microM, does not exert its negative inotropic effect by decreasing the calcium current. This mechanism may play a role at higher concentrations of the drug.     

蜂毒肽对豚鼠心室肌细胞钾电流和动作电位的影响(英文)

目的:研究蜂毒肽(Melittin,Mel)对豚鼠心室肌细胞钾电流和动作电位的影响.方法:全细胞膜片箝记录.结果:蜂毒肽可呈浓度依赖性促进延迟整流钾电流(I_k),在测定电压为40 mv时,0.05,0.1,0.2μmol·L~(-1)蜂毒肽分别使I_k从(295±109)增大到(371±142)(n=5 P<0.05),(467±180)(n=5,P<0.05),(552±248)pA(n=5,P<0.05).但药物在三个浓度时对内向整流钾电流(I_(k1))均无显著影响.蜂毒肽0.05,0.1,0.2 μmol·L~(-1)分别使动作电位APD_(50)由(520±55)减小到(459±91)(n=5,P>0.05),(385±102)(n=5,P<0.01),(281±81)ms(n=5,P<0.01),使APD_(90)由(613±96)减小到(536±93)(n=5,P>0.05),(467±96)(n=5,P<0.01),(354±95)ms(n=5,P<0.01).结论:蜂毒肽促进延迟整流钾电流,缩短动作电位时程.     

人重组生长激素对豚鼠离体心脏血流动力学及心肌细胞动作电位影响

目的 :观察人重组生长激素对正常豚鼠强心作用和对心室肌细胞动作电位的影响。方法 :采用langendorff离体心脏灌流法 ,悬浮玻璃电极法。结果 :注入人重组生长激素 10min后 ,左室最大收缩压 ,左室舒张末压 ,左室内压的最大升降速率的差值分别为 (6 .7± 5 .5 )kPa ,(0 .5 0± 0 .10 )kPa ,(139± 93)ps·s- 1,明显升高 ,心率差值为 (10± 16 )次·min- 1减慢 ,动作电位振幅值 (APA) ,超射 (OS) ,复极 5 0 %及 90 %水平的动作电位时程 (APD50 ,APD90 )分别为 (13.2± 6 )mv ,(9.6± 2 .2 )mv ,(84±2 9)ms,(90± 2 2 )ms ,显著延长 (均P <0 .0 1)。结论 :人重组生长激素对正常豚鼠具有正性肌力作用 ,可使动作电位时程延长     

Effects of goniopora toxin on the action potential and membrane currents of guinea-pig single ventricular cells

Summary The effects of Goniopora toxin (GPT), a polypeptide isolated from a coral, Goniopora spp., on action potential and membrane currents were studied in single ventricular cells of the guinea-pig using the whole-cell clamp technique with a single patch electrode. GPT at a concentration of 10 nmol/l prolonged the duration of the action potential without significant change in the resting membrane potential and action potential amplitudes. This prolongation became more evident at lower stimulus frequencies and persisted after washing with toxin-free solution. Tetrodotoxin (TTX, 1 mol/l), but not Co²⁺ (2 mmol/1), abolished the prolonged action potential. Under voltage-clamp conditions, a sustained inward current, not present in the control, followed the transient inward current during depolarizing pulses in the GPT-treated cells. The current-voltage relationship for the sustained inward current was much the same as that for the fast sodium current reported in rat single ventricular cells (Brown et al. 1981). Both the sustained and transient currents were abolished by the shift of holding potential in the direction of depolarization and reappeared after repolarization; the reappearance of the sustained current was much slower than that of the transient current. TTX but not Co²⁺ abolished both the sustained and transient inward currents. Calcium current and time-independent current were not affected by GPT. Time-dependent outward current induced by large depolarizing pulses was attenuated by GPT. From these results, it is suggested that GPT predominantly acts on the sodium channels in cardiac muscle, to give rise to a sustained sodium current which is responsible for the prolongation of action potential, and that the sodium channels modified by this toxin may reveal slow inactivation and slow recovery from inactivation as compared with those of unmodified sodium channels. Send offprint requests to I. Muramatsu     

Effects of lisinopril on electromechanical properties and membrane currents in guinea-pig cardiac preparations.

1. The effects of the angiotensin-converting enzyme inhibitor, lisinopril, were studied in guinea-pig atria and papillary muscles and in single isolated ventricular cells. 2. In isolated right atria, lisinopril (0.001-10 microM) decreased the amplitude and rate of the spontaneous contractions. In electrically driven left atria this negative inotropic effect was accompanied by a shortening of the time to peak tension and time for total contraction. 3. Lisinopril did not modify the electrophysiological characteristics of the ventricular action potentials recorded in papillary muscles perfused with normal Tyrode solution or elicited by isoprenaline in papillary muscles perfused with 27 mM K Tyrode solution. 4. In single ventricular cells, lisinopril (10 microM) had no effect on the inward L-type Ca2+ (ICa,L), the inward rectifier (IK1) or the delayed rectifier K+ currents (IK). However, it abolished the stimulation-dependent facilitation of the L-type Ca2+ current. 6. These results indicate that the negative inotropic effect of lisinopril cannot be explained by a decrease in Ca2+ entry through L-type channels and suggest that lisinopril may possibly act at an intracellular site to reduce contractile force.     

Effects of ervatamine chlorhydrate on cardiac membrane currents in frog atrial fibres.

1 The effects of a new alkaloid, ervatamine, on transmembrane currents of frog atrial fibres were studied by the double sucrose gap voltage clamp technique. 2 Ervatamine (2.8 x 10(-4) M) blocked the action potential without altering the resting membrane potential. 3 The alkaloid depressed the peak INa. The dissociation constant for the blocking effect of ervatamine on gNa fast was 2.35 X 10(-5) M with a one to one relationship between the drug molecule and the Na channel. Ervatamine did not alter the apparent equilibrium potential for Na, as well as the activation and inactivation parameters of gNa fast. This suggests that the alkaloid inhibitory effect on gNa can be attributed to a reduction in gNa. 4 Ervatamine prolonged the rate of reactivation of the Na system. It inhibited gNa in a frequency-dependent manner; this indicates that the alkaloid acts on open Na channels i.e. that the drug has to enter the channel or cross the membrane to produce the block. 5 Ervatamine inhibited Ina slow which occurs in Ca-free, tetrodotoxin-containing solutions and moderately decreased ICa which occurs in Na-free solutions. The drug increased the background K current (IK1) and did not alter the time-dependent K current (Ix1). 6 The present study shows that ervatamine is a good inhibitor of both fast and slow gNa. This drug also shares some common electrophysiological properties with antiarrhythmic drugs namely: the frequency-dependent inhibition of the fast gNa and the ability to slow the reactivation of the Na carrying system.     

Effects of an extract of Ginkgo biloba On the action potential and associated transmembrane ionic currents in mammalian cardiac myocytes: Inhibition of isoproterenol-induced chloride current

Ventricular myocytes of guinea pig heart were used to examine the effects of an extract of Ginkgo biolba (EGb 761) on the action potential and individual transmembrane ionic currents. Elecrophysiological events were recorded using the “whole-cell” configuration of the patch-clamp technique. A systematic analysis of the data revealed that EGb 761 (5-50 μg/ml) did not affect the normal action potential or the various ionic currents involved in its generation; i.e., fast inward sodium current (I_Na), inward calcium currents (I_CaT and I_CaL), delayed outward potassium current (I_K), inward rectifying potassium current (I_K1), and ATP-senstive potassium current (I_K-ATP) evoked by 2,4-dinitrophenol (2,4-DNP). However, EGb 761 (≥5 μg/ml) elicited a pronouced concentration-dependent and reversible inhibiton of isoproternol-induced Cl^? current (I_Cl), the maximal effect being observed at 50 μg/ml. This current may be significantly involved in sympathetic hyperactivity, hypoxia, and ischemia, pathophysiological conditions for which EGb 761 offers therapeutic benefit. The basic mechanism(s) underlying the inhibitory effect of EGb 761 on I_Cl and the constituent(s) of EGb 761 responsible for this action remain to be identified, but it seems clear, from results which showed that neither ginkgolide B (50–500 ng/ml) nor biolobalide (150–1,500 ng/ml) influenced this current, that a terpenoid constituent of EGb 761 is probably not involved.     

Effects of UTP on membrane current and potential in rat aortic myocytes

The electrophysiological effects of UTP on freshly isolated rat aortic myocytes were examined using the perforated patch clamp technique. Application of ,β-methylene ATP (β-meATP) and UTP, putative P2X and P2Y₂ or P2Y₄ purinoceptor agonists, induced transient and oscillatory inward currents, respectively. Experiments with Cl⁻ channel blockers and different external Cl⁻ concentrations demonstrated that the oscillatory current elicited by UTP is attributable to activation of Cl⁻ channels. The transient component elicited by β-meATP appeared to be responsible for a non-selective cationic current. With internal application of low-molecular-weight heparin, a blocker of inositol 1,4,5-trisphosphate (InsP₃), the oscillatory current elicited by UTP was abolished. The oscillatory current was activated in an all-or-none manner by UTP over the concentration range 0.1 and 1 μM and the frequency and amplitude were independent of the UTP concentration. Under current-clamp mode, UTP produced an oscillatory membrane potential. These results show that rat aortic myocytes have at least two types of P2 receptors. Activation of the P2Y receptor by UTP produces InsP₃, which releases Ca²⁺ from the store site. The resulting increase in intracellular Ca²⁺ concentration causes the oscillatory Cl⁻ current and the subsequent membrane potential changes.     

Comparison of the chronic and acute effects of amiodarone on the calcium and potassium currents in rabbit isolated cardiac myocytes.

1. The acute and chronic effects of amiodarone were studied on the transmembrane ionic currents in rabbit single ventricular myocytes at 35 degrees C by applying the whole-cell configuration of the patch-clamp technique. 2. Acute exposure to 1 and 5 microM amiodarone significantly reduced the amplitude (-53.9 +/- 3.9%, n = 5 and -64.0 +/- 2.0%, n = 3, P < 0.01), but chronic amiodarone treatment (i.p. 50 mg kg-1 day-1 for 3-4 weeks) changed neither the amplitude nor the kinetics of the inward calcium current. 3. Both acute superfusion with amiodarone (1 and 5 microM) and chronic amiodarone treatment significantly decreased the amplitude of the delayed rectifier outward potassium current (IK). 4. Acute application of amiodarone (1 and 5 microM) did not alter but chronic amiodarone treatment moderately depressed the transient outward current (Ito). 5. Neither the acute (1 and 5 microM) nor the chronic amiodarone treatment changed the magnitude of the inward rectifier potassium current (Ik1). 6. It is concluded that acute amiodarone application and chronic amiodarone treatment alter transmembrane ionic currents of ventricular myocytes differently. This may explain, at least in part, the marked differences in the cardiac electrophysiological effects observed after acute and chronic amiodarone treatment in patients.     

Differences in effects of forskolin and an analog on calcium currents in cardiac myocytes suggest intra- and extracellular sites of action.

The effects of forskolin (FO) and a water-soluble derivative of FO, L858051 (7 beta-desacetyl-7 beta-[gamma-(N-methylpiperazino)-butyryl] forskolin), were compared on calcium currents (ICa) studied by the whole-cell patch-clamp technique in frog ventricular cardiac myocytes. Both FO and L858051 increased ICa, with half-times of 160 +/- 20 sec and 343 +/- 22 sec, respectively. The stimulation was blocked by internal perfusion with inhibitors of protein kinase A. The EC50 for stimulation of ICa was 0.3 microM for FO and 1.0 microM for L858051. The maximal stimulated current was the same for both drugs, 20.3 microA/cm2 and 23.1 microA/cm2, respectively. Internal perfusion with 30-500 microM guanylyl 5'-imidodiphosphate [Gpp(NH)p] suppressed ICa stimulation by low concentrations of FO or L858051. This suppression was due to a rightward shift in the concentration-response curve, with increases in the EC50 values to 11.4 microM for FO and 28.4 microM for L858051. Isoproterenol (ISO) was ineffective in increasing ICa after the FO-stimulated ICa had been reduced by Gpp(NH)p and FO had been washed out. In contrast, after the L858051-stimulated current had been reduced by Gpp(NH)p, ISO stimulated ICa significantly. This stimulation was blocked by inhibitors of protein kinase A and was due to a positive effect of L858051 not shared by FO. A brief application of L858051 after Gpp(NH)p had blocked the ISO response restored the ISO response for at least 30 min. This effect was mimicked by internal perfusion with low concentrations of L858051. We conclude that the ability of brief exposure of L858051, but not FO, to restore the response to ISO after Gpp(NH)p is due to the accumulation of L858051 intracellularly, due to its hydrophilicity. Because internal L858051 and FO are very ineffective in stimulating adenylyl cyclase, whereas internal L858051 can restore the ISO response blocked by Gpp(NH)p, we propose that FO compounds can affect adenylyl cyclase at two sites, one site that is accessible only from the extracellular side that stimulates catalytic activity and another that is accessible from the intracellular side that increases beta-agonist efficacy in the presence of Gpp(NH)p.