Veratridine modification of Na current was examined in single dissociated ventricular myocytes from late-fetal rats. Extracellularly applied veratridine reduced peak Na current and induced a noninactivating current during the depolarizing pulse and an inward tail current that decayed exponentially (~ = 226 ms) after repolarization. The effect was quantitated as tail current amplitude, /tail (measured 10 ms after repolarization), relative to the maximum amplitude induced by a combination of 100 IzM veratridine and 1 ~M BDF 9145 (which removes inactivation) in the same cell. Saturation curves for Itail were predicted on the assumption of reversible veratridine binding to open Na channels during the pulse with reaction rate constants determined previously in the same type of cell at single Na channels comodified with BDF 9145. Experimental relationships between veratridine concentration and Itau confirmed those predicted by showing (a) haftmaximum effect near 60 ~M veratridine and no saturation up to 300 ~M in cells with normally inactivating Na channels, and (b) haft-maximum effect near 3.5 p,M and saturation at 30 ~M in cells treated with BDF 9145. Due to its known suppressive effect on single channel conductance, veratridine induced a progressive, but partial reduction of noninactivating Na current during the 50-ms depolarizations in the presence of BDF 9145, the kinetics of which were consistent with veratridine association kinetics in showing a decrease in time constant from 57 to 22 and 11 ms, when veratridine concentration was raised from 3 to 10 and 30 I.LM, respectively. As predicted for a dissociation process, the tail current time constant was insensitive to veratridine concentration in the range from 1 to 300 o,M. In conclusion, we have shown that macroscopic Na current of a veratridine-treated cardiomyocyte can be quantitatively predicted on the assumption of a direct relationship between veratridine binding dynamics and Na current and as such can be successfully used to analyze molecular properties of the veratridine receptor site at the cardiac Na channel.

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1. The inotropic and electrophysiological effects of aconitine were measured in the isolated, isometrically contracting guinea-pig papillary muscle during the prearrhythmic phase of alkaloid action. 2. In muscles stimulated continually at 1 Hz, 1 mumol/l aconitine produced a positive inotropic effect that reached 38 +/- (SEM) 9% immediately before the onset of arrhythmia (n = 3). 3. If aconitine (0.5 mumol/l) was applied to non-stimulated (resting) muscles for 30 min and 1-Hz stimulation resumed thereafter, the arrhythmia occurred after 724 +/- 101 beats. Prolongation of the rest exposure to 2 h did not significantly diminish the number of prearrhythmic beats. Thus, the onset of aconitine action is critically determined by muscle activity (rather than by time), and a 30-min aconitine application to the resting muscle suffices for complete equilibration of the tissue. 4. Using the preequilibration-at-rest procedure, the positive inotropic effect of aconitine (0.25 - 4 mumol) was found (a) to be absent in the rested-state contraction, (b) to grow with both number of subsequent beats and alkaloid concentration, and (c) to reach a similar prearrhythmic maximum at all concentrations. This maximum amounted to about 1/4 of the maximum positive inotropic effect of dihydroouabain. It was not influenced by reserpine pretreatment of the guinea pig. 5. Aconitine (1 mumol/l) delayed the repolarization phase of the action potential by establishing a secondary plateau at approximately -60 mV. This effect paralleled the positive inotropic effect and, like the positive inotropic effect, was abolished by 10 mumol/l tetrodotoxin (TTX). In partially depolarized muscles ([K]0 = 24 mmol/l) aconitine (8 mumol/l) produced a TTX-sensitive increase in amplitude and rate of rise of the rested-state contraction; this indicates a voltage-dependent effect on some resting Na channels. 6. While delaying the late repolarization phase, aconitine markedly shortened the early repolarization at levels positive to -40 mV, reduced the overshoot and decreased the maximum rat of depolarization of the action potential. Slow action potentials ([K]0 = 24 mmol/l; 10 mumol/l TTX) were insensitive to aconitine. 7. We conclude that the well known property of aconitine to prolong the Na influx during the action potential leads to a positive inotropic effect, thus confirming the importance of Na influx for the regulation of myocardial contractility. The exact mechanism of an additional effect by which aconitine reduces the overshoot and shortens the plateau phase of the action potential awaits further study.

In the isolated papillary muscle of the guinea pig veratridine produces an increase of the force of contraction by increasing the rate of force development. Time to peak force is slightly reduced, whereas relaxation time is markedly prolonged. Threshold, half-maximally and maximally effective concentrations for the positive inotropic effect are 0.1, 0.4 and 1.6 muM, respectively. 2. The positive inotropic effect of the maximally effective concentration of veratridine amounts to 68% of the maximum positive inotropic effect of dihydro-ouabain tested on the same muscle (N = 12). 3. Veratridine prolongs the action potential (AP) by delaying repolarization. The effect is concentration-dependent (range: 0.4--3.2 muM); it requires 1--2 hrs of maintained exposure to reach a steady state and is only slowly reversible upon removal of the drug. A concentration causing a nearly maximal positive inotropic effect (0.8 muM) does not affect resting potential or rate of rise of the AP; the overshoot is slightly depressed. 4. Tetrodotoxin (5--16 muM) reversibly inhibits both AP prolongation and positive inotropic effect of veratridine by shifting the concentration-effect curves for these effects to higher concentrations of veratridine. It also prevents veratridine-induced spontaneous activity. 5. Dihydro-ouabain or reduction of [K]o below 5.9 mM augument the positive inotropic effect of veratridine, while the interaction between veratridine and noradrenaline is additive. 6. The positive inotropic effect of 1.6 muM veratridine declines progressively when the contraction frequency is reduced below 0.5 Hz; rested-state contractions (at 0.004 Hz) are not increased by 1.6 muM veratridine. 7. It is concluded that (a) veratridine delays repolarization by prolonging the Na permeability component which is mediated by the fast Na channels; (b) this specific sarcolemmal effect of veratridine is the sole cause for its positive inotropic action by effecting an increase of [Na]i which probably leads to a subsequent increase of Ca uptake.

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