Modeling the effect of Kv1.5 block on the canine action potential

A wide range of ion channels have been considered as potential targets for pharmacological treatment of atrial fibrillation. The Kv1.5 channel, carrying the I_Kur current, has received special attention because it contributes to repolarization in the atria but is absent or weakly expressed in ventricular tissue. The dog serves as an important animal model for electrophysiological studies of the heart and mathematical models of the canine atrial action potential (CAAP) have been developed to study the interplay between ionic currents. To enable more-realistic studies on the effects of Kv1.5 blockers on the CAAP in silico, two continuous-time Markov models of the guarded receptor type were formulated for Kv1.5 and subsequently inserted into the Ramirez-Nattel-Courtemanche model of the CAAP. The main findings were: 1), time- and state-dependent Markov models of open-channel Kv1.5 block gave significantly different results compared to a time- and state-independent model with a downscaled conductance; 2), the outcome of Kv1.5 block on the macroscopic system variable APD₉₀ was dependent on the precise mechanism of block; and 3), open-channel block produced a reverse use-dependent prolongation of APD₉₀. This study suggests that more-complex ion-channel models are a prerequisite for quantitative modeling of drug effects.     

In silico study on the effects of IKur block kinetics on prolongation of human action potential after atrial fibrillation-induced electrical remodeling

Pharmacological treatment with various antiarrhythmic agents for the termination or prevention of atrial fibrillation (AF) is not yet satisfactory. This is in part because the drugs may not be sufficiently selective for the atrium, and they often cause ventricular arrhythmias. The ultrarapid-delayed rectifying potassium current (I(Kur)) is found in the atrium but not in the ventricle, and it has been recognized as a potentially promising target for anti-AF drugs that would be without ventricular proarrhythmia. Several new agents that specifically block I(Kur) have been developed. They block I(Kur) in a voltage- and time-dependent manner. Here we use mathematical models of normal and electrically remodeled human atrial action potentials to examine the effects of the blockade kinetics of I(Kur) on atrial action potential duration (APD). It was found that after AF remodeling, an I(Kur) blocker with fast onset can effectively prolong APD at any stimulus frequency, whereas a blocker with slow onset prolongs APD in a frequency-dependent manner only when the recovery is slow. The results suggest that the voltage and time dependence of I(Kur) blockade should be taken into account in the testing of anti-AF drugs. This modeling study suggests that a simple voltage-clamp protocol with a short pulse of approximately 10 ms at 1 Hz may be useful to identify the effective anti-AF drugs among various I(Kur) blockers.     

Fast frequency-dependent block of action potential upstroke in rabbit atrium by small local anesthetics.

Local anesthetics depress the rapid depolarizing phase of atrial action potentials in a frequency-dependent manner. Several local anesthetics were tested in this study at doses that only slightly depressed atrial action potential upstrokes at the spontaneous rate near 2 Hz, but that produced marked reductions in atrial fiber upstrokes when the drive rate was increased to 5 Hz. This frequency-dependent blocking action, especially the rate of development of the block during the 5-Hz stimulus train, is compared for several local anesthetics that have a wide range of lipid solubilities. Although the rate of block development does not correlate well with lipid solubility, it does correlate with molecular weight or size of the local anesthetic molecule. Smaller local anesthetics give faster frequency-dependent blocking. The beta-blocker propranolol also induces a frequency-dependent block of action potential upstroke, with the speed of such block development being predictable on the basis of the molecular weight of propranolol. The design of fast frequency-dependent blockers by using the criterion of smaller molecular size represents an important new structure-activity relation that may very well help in the design of better antifibrillatory drugs.     

Prostaglandins do not modulate the positive chronotropic and inotropic effects of sympathetic nerve stimulation and injected norepinephrine in the isolated blood perfused canine atrium

In isolated canine atrium, perfused with blood from a donor dog, the infusions of both prostaglandins (PG)I₂ and E₂ (0.1–1 μg/min) into the sinus node arterial cannula neither altered the sinus rate and developed tension nor the positive chronotropic and inotropic responses elicited by either electrical stimulation or by injected norepinephrine. Infusion of arachidonic acid (10–100 μg/min), a precursor of PGs, or indomethacin (15–20 μg/min), an inhibitor of PG synthesis, into the sinus node arterial cannula also failed to alter the increase in sinus rate or developed tension produced by either adrenergic stimulus in the isolated atria. When arachidonic acid, 100–300 μg/kg or PGI₂, 1 μg/kg, were injected into the jugular vein of the donor dog, they produced a fall in systemic blood pressure; this effect of arachidonic acid but not of PGI₂ was abolished by indomethacin, 1 mg/kg. During administration of either arachidonic acid or indomethacin to the donor dog, the positive chronotripic and inotropic responses to adrenergic stimuli in the isolated atria also remained unaltered. These data indicate that PGs do not modulate adrenergic transmission in the blood perfused canine atrium.     

Vagus effect on action potentials of rest-potentiated contractions in the isolated left atrium of the rabbit

After a period of rest action potentials in constantly driven preparations of left rabbit atria show a marked change in configuration. After the upstroke an early repolarization takes place which is followed by a prolonged phase of secondary depolarization. This depolarization is strongly suppressed by vagal stimulation. Frequency-response characteristics of this electrotropic effect were obtained by stimulating the vagal supply of the preparation with frequencies in the range from 1.0 to 40.0 s-1 and compared with those of the constantly driven preparation. In most cases studied the frequency-response curves for the post rest action potential are steeper in rise and shifted to the left. By placing a series of vagal stimulations at different moments into the resting interval vagal effects could be composed to reflect the time course of transmitter action in a non beating preparation. It was shown that such time course is not essentially altered compared with that of a driven preparation. From a mathematical treatment of the frequency-response relations it is concluded that post rest action potentials show a higher sensitivity towards the transmitter action and that the amount of transmitter liberated after a period of rest may be increased. Possible explanations for this behaviour are proposed.     

The effect of stimulation of the adrenergic receptors on the frequency depression of the temporal parameters of the action potential in the right atrium of rats]

At a stimulation rate of 1 Hz, activation of alpha-adrenoreceptors prolonged the AP duration at 10%, 50%, and 90% repolarisation at 10(-7), 10(-6) M in the rat isolated right atria, but shortened it at a higher concentration of 10(-5) M. The frequency-induced depression of the AP duration became more evident in cardiomyocytes stimulated by 10(-7), 10(-6) M and less obvious at 10(-5) M of alpha-adrenoagonist. Activation of alpha-adrenoreceptors by isoprenalin shortened the AP duration and enhanced the stimulation-rate-dependent changes in the atrial AP configuration.     

Leukotriene C4 action and metabolism in the isolated perfused bullfrog heart

The effects of leukotrienes (LTs) have been widely studied in the isolated perfused mammalian heart; however, little is known about the effect or metabolism of LTs in the isolated bullfrog heart. Isolated perfused bullfrog hearts were administered randomized doses of LTC4, LTD4, or LTE4. The cardiac parameters of heart rate, developed tension, and its first derivative (dT/dt) were recorded. LTC4 was the most potent of the leukotrienes tested in eliciting positive inotropic effects. LTD4 and LTE4 were equally effective but about one order of magnitude less potent than LTC4. None of the LTs showed any chronotropic effects in this preparation. A series of [3H]LTC4 metabolism experiments were carried out using whole perfused hearts and minced bullfrog heart tissue. Isolated perfused bullfrog hearts administered [3H]LTC4 converted significant amounts to [3H]LTD4, and to a lesser degree, [3H]LTE4, during the 6-min course of collection. Both minced atrial and ventricular tissue converted [3H]LTC4 to radioactive metabolites that co-migrated with authentic LTD4 and LTE4 standards. In both tissues, the major product was [3H]LTD4, with smaller amounts of [3H]LTE4 produced. The atrium converted significantly more [3H]LTC4 to its metabolites than did the ventricle. The metabolism of [3H]LTC4 to [3H]LTD4 by both tissues was virtually abolished in the presence of serine borate. Cysteine had no effect on [3H]LTE4 production. The data in this study demonstrate that leukotrienes have the opposite inotropic effect on the heart when compared with mammals. Also in contrast to mammals, frogs metabolize LTC4 to a less potent compound and may use the LTC4 to LTD4 conversion as a mechanism of LTC4 inactivation.     

Mechanism of action of arachidonic acid in the isolated perfused rat heart

The purpose of this study was to elucidate the mechanism of action of arachidonic acid in the isolated rat heart perfused with Krebs solution at a constant flow. Administration of arachidonic acid, 3.3-33 nmol, into the heart caused a small transient increase followed by a pronounced decrease in coronary perfusion pressure and increased myocardial tension, heart rate, and the output of prostaglandins (6-keto-PGF1 alpha, PGE2, and PGF2 alpha). Administration of structurally similar fatty acids, dihomo-gamma-linolenic acid, and 8,14,17-eicosatrienoic acid, produced vasoconstriction and decreased myocardial tension without affecting heart rate or the output of prostaglandins. Infusion of PGI2, PGF2 alpha, or PGE2 produced coronary vasodilation and increased myocardial tension, whereas PGF2 alpha increased heart rate, an effect which was not prevented by propranolol. Indomethacin blocked the effect of arachidonic acid on myocardial tension and heart rate, but only reduced the duration of coronary vasodilation. The initial component of arachidonic acid induced coronary vasodilation which was unaffected by indomethacin and also remained unaltered during the infusion of three structurally dissimilar lipoxygenase inhibitors, eicosatetraynoic acid, nordihydroguaiaretic acid, and 1-phenyl-3-pyrazolidone. Indomethacin did not alter the effects of the exogenously administered prostaglandins on perfusion pressure or myocardial tension; however, it blocked the effect of PGF2 alpha on heart rate. The effect of arachidonic acid or PGF2 alpha to increase heart rate was not blocked by thromboxane synthetase inhibitors, imidazole, or OKY-1581. We conclude that the cardiac effects of arachidonic acid are mediated primarily through its conversion to cyclooxygenase products.(ABSTRACT TRUNCATED AT 250 WORDS)     

Jejunal factor stimulating insulin release in the isolated perfused canine pancreas and jejunum.

In the present study using an isolated perfused preparation of canine jejunum and pancreas, an insulin-releasing factor was found in the venous effluent of the jejunum. Insulin secretion by the pancreas rose twofold after 10% glucose was infused in the lumen of the jejunum and remained at a high level even after the stimulus was discontinued. No modification of the exocrine pancreatic secretion occurred during the insulin release, and therefore it seems unlikely that gastrin, secretin or cholecystokinin-pancreozymin were released by the jejunal mucosa. In control experiments the values of hyperglycaemia observed previously and intraluminal hyperosmolarity were tested: at these levels, they did not affect insulin secretion. The nature of this intestinal insulin-releasing factor remains unknown however, but may be identifiable when intestinal hormones in blood can be assayed reliably.     

Heterogeneity of action potential durations in isolated mouse left and right atria recorded using voltage-sensitive dye mapping

An imaging system for di-4-ANEPPS (4-[beta-[2-(di-n-butylamino)-6-naphthylvinyl]pyridinium]) voltage-sensitive dye recordings has been adapted for recording from an in vitro mouse heart preparation that consists of both atria in isolation. This approach has been used to study inter- and intra-atrial activation and conduction and to monitor action potential durations (APDs) in the left and right atrium. The findings from this study confirm some of our previous findings in isolated mouse atrial myocytes and demonstrate that many electrophysiological properties of mouse atria closely resemble those of larger mammals. Specifically, we made the following observations: 1) Activation in mouse atria originates in the sinoatrial node and spreads into the right atrium and, after a delay, into the left atrium. 2) APD in the left atrium is shorter than in the right atrium. 3) Sites in the posterior walls have longer APDs than sites in the atrial appendages. 4) Superfusion of this preparation with 4-aminopyridine and tetraethylammonium resulted in increases in APD, consistent with their inhibitory effects on the K+ currents known to be expressed in mouse atria. 5) The muscarinic agonist carbachol shortened APD in all areas of the preparation, except the left atrial appendage, in which carbachol had no statistically significant effect on APD. These results validate a new approach for monitoring activation, conduction, and repolarization in mouse atria and demonstrate that the physiological and pharmacological properties of mouse atria are sufficiently similar to those of larger animals to warrant further studies using this preparation.     

Angiotensin II and norepinephrine after indomethacin in isolated perfused canine kidneys. Tachyphylaxis vs. modulator effect of prostaglandins

High levels of radioimmunoassayable PGE₂ were measured in the perfusate of isolated kidneys. Indomethacin inhibited PGE₂ release in this system. Small reductions in the pressor effects of norepinephrine (NE) were associated with increasing perfusate levels of PGE₂; a large increase in the pressor effect of NE followed additions of indomethacin and reductions in perfusate PGE₂ levels. A marked reduction in pressor responsiveness to angiotensin II (AII) was measured in the isolated kidney which could not be prevented or reversed by indomethacin. It is believed that tachyphylaxis was responsible for the marked reduction in pressor responsiveness to AII and that this is independent of alterations in prostaglandin metabolism. However prostaglandins appeared to modulate the pressor effects of AII as they did NE in the isolated perfused kidney.     

Muscarinic type 1 receptors mediate part of nitric oxide's vagal facilitatory effect in the isolated innervated rat right atrium.

We investigated whether vagal cardiac cholinergic facilitation by nitric oxide (NO) is mediated by cardiac muscarinic receptor subtypes in the vagally innervated rat right atrium in vitro. Experiments were carried out in the presence of atenolol (4 microM). The right vagus was stimulated at 4, 8, 16, 32 Hz; pulse duration 1 ms at 20 V for 20s; vagal postganglionic activation was achieved using nicotine (0.1, 0.3, 0.5, 1mM) and the effect on cardiac interval (ms) assessed. Pirenzepine (1 microM), a M1 antagonist, attenuated vagally induced increase in cardiac interval. L-Arginine (0.34 mM) superfused with pirenzepine failed to reverse this attenuation, however, L-arginine applied alone reversed the reduction vagal cardiac slowing. Similarly, sodium nitroprusside (10 microM) applied alone, and not together with pirenzepine, was able to reverse the attenuation of vagal effects caused by pirenzepine. Synthetic MT7 (1 nM) toxin, a selective M1 antagonist confirmed these results. M3 antagonism using para-fluorohexahydrosiladifenidol (p-F-HHSiD) (300 nM) and M4 antagonism with PD 102807 (200 nM) did not affect the vagally induced increase in cardiac interval. Nicotine induced increase in cardiac interval was not altered by pirenzepine. These results show that antagonism of M1 receptors on cardiac vagal preganglionic fibres reduces vagal efficacy which can be recovered by either a nitric oxide synthase substrate or a NO donor.     

The effects of bradykinin on electrobiological phenomena in isolated right atrium of the rat.

Bradykinin induces or accelerates the already present electrobiological activity in isolated right atrium of rats. Acceleration of the electrobiological activity induced by bradykinin decreases with increase of initial activity. Bradykinin disturbs transiently the course of the repolarization period in action potential. It may be one of the mechanisms of arrhythmia development caused by bradykinin.