Changes in intracellular Na+ during Na,K pump inhibition in sheep cardiac tissues.

The differences in inotropic and toxic sensitivities of cardiac ventricular tissue to cardiac glycosides were investigated in order to determine whether or not the reportedly greater sensitivity of Purkinje fibers compared to myocardium is the result of a fundamentally different response to Na,K pump inhibition. I measured the changes in intracellular Na+ activity (aiNa) in the two tissue types simultaneously during exposure to actodigin (4.0 microM) and ouabain (0.5 microM) under quiescent conditions. A sheep papillary muscle and a Purkinje fiber from the same heart were placed in an experimental chamber and measurements of aiNa from both were obtained with Na+ -sensitive microelectrodes. In five experiments in which all electrode impalements were successfully maintained, actodigin caused similar changes in aiNa in the two tissues (from 7.2 +/- 1.0 mM in control to 12.9 +/- 1.9 mM in the presence of drug in papillary muscles compared to 7.3 +/- 0.3 mM in control and 13.2 +/- 1.0 mM in Purkinje fibers; means +/- S.E.M.). After washout, exposure to ouabain increased aiNa in both papillary muscles and in Purkinje fibers (from 7.2 +/- 0.7 mM in control and 16.2 +/- 1.4 mM during exposure to drug in papillary muscles compared to 7.4 +/- 0.3 mM and 14.9 +/- 0.8 mM in Purkinje fibers). In fact, ouabain caused a greater increase of aiNa in papillary muscles than in Purkinje fibers (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)     

Alternans of action potential duration after abrupt shortening of cycle length: differences between dog Purkinje and ventricular muscle fibers

The purpose of this study was to determine whether the alternans of action potential duration (APD) occurring in Purkinje and ventricular muscle fibers after an abrupt shortening of cycle length can be explained by the two factors controlling the cycle length-dependent APD changes (i.e., restitution and memory effect). Action potentials were recorded simultaneously from dog Purkinje fibers and ventricular muscle fibers using conventional microelectrode techniques. APD change during alternans was dependent on the preceding diastolic interval in the same manner as during restitution in Purkinje fibers but not in ventricular muscle fibers. The course of memory change was not affected by the presence of alternans in either fiber type. In Purkinje fibers, APD alternans was attenuated by a Ca2+ channel blocker, nisoldipine (2 X 10(-6) M), and augmented by a Ca2+ channel agonist, Bay K 8644 (3 X 10(-8) M). These effects were attributed to the changes in the kinetics and the amplitude of restitution. In ventricular muscle fibers, APD alternans was always preceded and accompanied by alternans of action potential shape. Alternans of both action potential shape and APD was suppressed by nisoldipine (2 X 10(-6) M) and attenuated by Bay K 8644 (3 X 10(-8) M). These results show that in Purkinje fibers, APD during alternans can be explained by restitution and memory effect. However, in ventricular muscle fibers, the mechanism of APD alternans is linked to factors controlling action potential shape. These findings are compatible with the hypothesis that APD alternans in Purkinje fibers depends on the differences in the recovery of membrane currents generated by the preceding action potential and in ventricular muscle fibers on the differences in the concentration and/or handling of intracellular calcium.     

Effects of bepridil on the electrophysiologic properties of isolated canine and rabbit myocardial fibers

Bepridil hydrochloride is a relatively new calcium antagonist which appears to have a complex pharmacologic profile, but its concentration-response characteristics with respect to its electrophysiologic properties of varying concentrations (0.1 to 10.0 micrograms/ml) of the drug were therefore determined in rabbit and canine myocardial fiber preparations in vitro by standard microelectrode techniques. The following were measured: sinus cycle length (SCL), action potential amplitude (APA), maximum diastolic potential (MDP), threshold potential (TP), slope of phase 4 depolarization, action potential duration (APD), and dV/dtmax of phase O depolarization (Vmax) in rabbit sinoatrial (SA) node. Also measured were APA, membrane resting potential (MRP), Vmax, APD at 50% and 90% repolarization (APD50 and APD90), and effective refractory period (ERP) in rabbit atria and canine Purkinje fibers and ventricular muscle. At the lowest concentrations bepridil selectively prolonged SCL by reducing the slope of phase 4 and decreased APA and MDP in a concentration-dependent manner in the sinus node. At higher concentrations, bepridil exerted additional effects in producing concentration-dependent decreases in APA and Vmax in rabbit atria and in canine Purkinje fibers and ventricular muscle. During superfusion with 1.0 micrograms/ml bepridil, Vmax fell by 22.2% (p less than 0.05) in Purkinje fibers and by 11.8% (NS) in ventricular muscle; at 10.0 micrograms/ml, Vmax fell by 46.5% (p less than 0.01), respectively. The depression of Vmax was frequency dependent. There was a differential effect of bepridil on repolarization in Purkinje fibers as compared to that in ventricular muscle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison of rubidium-86 and potassium-42 fluxes in rat aorta

Potassium transport has frequently been assessed by measurement of 86Rb. However, recent reports indicated that the K ion channels are selective to K over Rb. Therefore, the purpose of this study was to evaluate whether the basal and stimulated fluxes of Rb and K were equivalent in rat aorta in the absence of endothelium. The ouabain-sensitive (active) uptake of 86Rb and 42K was similar. However, the basal 86Rb efflux was only 80% of the 42K efflux. Norepinephrine and KCl depolarization stimulated 86Rb and 42K effluxes via a calcium-dependent process. The stimulated 86Rb efflux ranged from 56 to 74% of the 42K efflux. Diltiazem reduced the KCl-stimulated 86Rb and 42K effluxes. The 86Rb efflux was 82% of the 42K efflux in the presence of KCl plus diltiazem, similar to that under basal conditions. Substitution of Rb for K in the incubation solution was associated with a marked increase in spontaneous contractile activity. There was no change in the norepinephrine concentration required for a 50% stimulation of contraction or 86Rb efflux from Rb-loaded tissues. We conclude from these studies that the basal and calcium-activated potassium channels are selective for K over Rb and therefore 86Rb fluxes quantitatively underestimate that of 42K. However, 86Rb is an appropriate substitute for the measurement of active K transport.     

Effects of norepinephine, calcium, and rate of discharge on 42K movements in canine cardiac Purkinje fibers

We studied the effect of norepinephrine, calcium concentration, and rate of discharge in the presence of different [Ca2+]0 on radioactive potassium movements in cardiac Purkinje fibers. The following results were obtained: (1) norepinephrine increases potassium uptake in quiescent fibers and in fibers driven at constant rate, but more in the latter; (2) norepinephrine also increases potassium uptake in quiescent fibers depolarized at the plateau; (3) increasing [Ca2+]0 increases potassium uptake in fibers driven at constant rate; (4) increasing [Ca2+]0 may decrease K+ uptake in quiescent fibers; (5) increasing [Ca2+]0 decreases the rate of loss of tissue radioactivity in quiescent fibers and increases it in a driven fiber; (6) increasing the driving rate increases potassium uptake in low and high [Ca2+]0; (7) high [Ca2+]0 increases K+ uptake, especially at low rates; (8) norepinephrine is less effective in increasing K+ uptake in the presence of a high [Ca2+]0. We conclude that: (a) norepinephrine increases potassium uptake by different mechanisms; (b) calcium affects potassium movements when it is allowed to enter the cell, presumably by affecting potassium conductance; (c) the effect of an increased rate of discharge on K+ uptake may involve stimulation of active K+ uptake and may include a calcium-dependent component which is largest at high [Ca2+]0 and a slow rate of drive; (d) simultaneous application of two interventions results in a summation that is smallest when one of the mechanisms is already substantially activated.     

Ouabain sensitivity of the Na, K-ATPase activity from single bovine cardiac Purkinje fiber and adjacent papillary muscle

When assessed electrophysiologically, the sensitivity of Purkinje fibers to the toxic effects of ouabain is greater than that of ventricular muscle fibers. We have attempted to determine in vitro whether this is a reflection of a differential sensitivity of the Na⁺, K⁺-ATPase activity. Purkinje fibers and adjacent papillary muscle were homogenized in a medium which prevented the formation of impermeable membrane vesicles, thus preventing ion accumulation and allowing both sides of the membrane to be exposed to ouabaain, substrate, and cofactors. Under these conditions the Na⁺, K⁺-ATPase activity from Purkinje fibers was not more sensitive to ouabain compared to the activity from adjacent papillary muscle. The results suggest that factors other than inherent ouabain sensitivity of the Na⁺, K⁺-ATPase must underlie the apparent differential ouabain-Na⁺, K⁺-ATPase interaction in vivo.     

Effects of propafenone on sinus nodal and ventricular automaticity: in vitro and in vivo correlation

The electrophysiologic effects of the new antiarrhythmic drug, propafenone, were evaluated in anesthetized closed-chest dogs and on isolated cardiac tissues with the microelectrode technique. Propafenone (2 to 4 mg/kg intravenously) had no effect on sinus rate or on sinus nodal recovery time, but caused a dose-dependent significant decrease in the rate of idioventricular rhythm and increased the duration of ventricular overdrive suppression in dogs (n = 8) with complete atrioventricular block. On isolated canine Purkinje fibers (n = 8) manifesting automaticity with resting membrane potential less negative than -70 mV, propafenone reduced the slope of phase 4 depolarization and reduced the rate of automatic impulse initiation in a concentration-dependent manner (10(-6) M-4.10(-5) M). At these concentrations, propafenone had no effect on rabbit sinus nodal automaticity (n = 8) or on sinoatrial conduction. However, significant depression of sinus nodal automaticity occurred with propafenone concentrations above 5.10(-6) M in the presence of cholinergic or complete autonomic blockade with atropine (10(-6) M) and propranolol (5.10(-5) M). Propafenone caused a concentration-dependent decrease in the disparity of Purkinje fiber-ventricular muscle action potential duration (APD), mainly by shortening Purkinje fiber APD. We conclude: that propafenone suppresses idioventricular rhythm in the intact dog, most likely by depressing Purkinje fiber automaticity; the depressant effect of propafenone on sinus nodal automaticity is evident only during cholinergic receptor blockade; and the antiarrhythmic properties of propafenone may include removal of APD disparity by selective shortening of Purkinje fiber and not of ventricular muscle APD.     

Stimulation of sugar transport in rat soleus muscle by prolonged cooling at 0 °C

Summary The uptake of D-xylose by isolated rat soleus muscle (measured at 37 °C) was stimulated by prolonged cooling at 0 °C. The effect of cooling reached a maximum value after 3 h and was reversed on rewarming; reversal was temperature-dependent. Cooling stimulated xylose uptake sub-maximally compared with the effect of insulin (100 U/l). Xylose uptake in cooled muscle was further stimulated by insulin, but not by anoxia. The effect of cooling and its reversal were still demonstrable in the presence of ouabain (1 mmol/l), or when unidirectional efflux of calcium and magnesium from the muscle was induced by EDTA (5 mmol/l). The ionophore, A23187 (2.5 mg/l), depressed the effect of cooling in the presence of EDTA but not in the presence of EGTA. It is concluded that cooling disrupts an intracellular magnesium-pump and that muscle sugar transport is consequentially stimulated through an increase in cytoplasmic magnesium.     

Electrophysiology and ultrastructure of canine subendocardial Purkinje cells isolated from control and 24-hour infarcted hearts

Ventricular arrhythmias that accompany myocardial infarction in dogs may be secondary to the altered electrophysiological properties of the subendocardial Purkinje fibers that survive 24 hours after the coronary occlusion. To better understand the ionic mechanisms that underlie the altered electrical activity of these fibers, we have dispersed, using an enzymatic technique, Purkinje cells from the subendocardium of the infarcted ventricle (IZPCs) and compared their electrical and structural properties to Purkinje cells dispersed from fiber strands (SPCs) and from the subendocardium of the noninfarcted ventricle (NZPCs). Ultrastructural analysis of these cells shows that IZPCs contain an increased number of lipid droplets when compared with the SPCs and NZPCs. In addition, transmembrane action potentials of IZPCs have reduced resting potentials, action potential amplitudes, and upstroke velocity and are increased in duration when compared with either SPCs or NZPCs. Input resistance of IZPCs is increased over that measured in control cells (SPCs and NZPCs). Furthermore, the time course of the process of electrical restitution of action potential duration is altered in IZPCs with long action potentials. Finally, using K+-sensitive microelectrode techniques, we have determined that intracellular free K+ activity (aKi) in IZPCs (93.7 +/- 15 mM) is not significantly different from control aKi measurements (SPC, 106 +/- 13 mM; NZPC, 103 +/- 12 mM). Thus a reduction in aKi does not provide a basis for the reduced resting potentials observed in IZPCs. By studying the relation between the resting potential and log [K+]o we determined that in IZPCs with reduced resting potentials, there is a significant increase in the PNa/PK ratio when compared with control. In summary, to better understand the cellular basis of ventricular arrhythmias postinfarction, we have developed a single cell model that will allow for more rigorous electrophysiological studies of the specific ionic currents that underlie the abnormal electrophysiology.     

Dantrolene sodium: Effects on isolated cardiac tissues

The effects of dantrolene sodium on dog Purkinje fibers, cat atrial and ventricular muscles were studied. Action potential duration was significantly increased and contractility was significantly decreased by dantrolene in all three types of tissue. The plateau phase of Purkinje fiber and occasionally atrial action potential was slightly depressed. Dantrolene sodium had no significant effect on resting membrane potential, action potential amplitude or upstroke velocity of phase 0. The negative inotropic effects were most pronounced in Purkinje fibers, followed by atrial muscle while papillary muscles were least sensitive. Contractile force of Purkinje fibers was decreased by relatively the same amount at all frequencies of stimulation. At faster rates, atrial and ventricular muscle contractility was depressed relatively more than at slower rates. Slow response action potentials in cat papillary muscles were diminished slightly, but this effect was not significant. All drug effects took 10 to 15 min to develop, reached a steady state after 30 to 40 min, and were irreversible upon washout. Increasing the extracellular calcium concentration reversed the dantrolene-induced changes. These findings suggest that effects of dantrolene are mediated in part by a decrease in the intracellular free calcium concentration in cardiac tissues.     

Enhancement of triggered activity in ischemic Purkinje fibers by ouabain: a mechanism of increased susceptibility to digitalis toxicity in myocardial infarction

Enhanced susceptibility to toxic arrhythmias by digitalis administration has been reported in clinical and experimental myocardial infarction. To investigate the mechanism responsible for this phenomenon, the effects of superfusion with normal Tyrode's solution and superfusion with Tyrode's solution containing 4 X 10(-8)M of ouabain in ischemic Purkinje fibers were compared. Ischemic Purkinje fibers of small endocardial preparations from 1 day old myocardial infarcts in 18 dogs were used for the study. During control conditions, these endocardial preparations demonstrated delayed afterdepolarizations and triggered activity. Superfusion with normal Tyrode's solution resulted in a gradual increase in maximal diastolic potential and action potential amplitude, a decrease in delayed afterdepolarizations amplitude and slowing and termination of triggered activity. Superfusion for 90 minutes with Tyrode's solution containing ouabain resulted in: 1) an increase in the magnitude of delayed afterdepolarizations in preparations demonstrating subthreshold delayed afterdepolarizations, 2) sustainment of triggered activity in preparations showing nonsustained triggered activity, and 3) shortening of cycle lengths of the triggered activity in preparations demonstrating sustained triggered activity before superfusion with ouabain. These effects occurred despite the gradual increase in maximal diastolic potential and action potential amplitude. Superfusion of normal Purkinje fibers with Tyrode's solution containing 4 X 10(-8)M of ouabain for 90 minutes did not result in delayed afterdepolarizations or triggered activity. Thus, ouabain at a concentration that has no toxic effect on normal Purkinje fibers may enhance arrhythmias in ischemic Purkinje fibers by increasing the magnitude of delayed afterdepolarizations and enhancing triggered activity.     

Calcium and magnesium transport by in situ mitochondria: electron probe analysis of vascular smooth muscle

The extent, time course, and reversibility of mitochondrial Ca2+ uptake secondary to cellular Ca2+ influx stimulated by massive Na+ efflux were evaluated by electron probe microanalysis of rabbit portal vein smooth muscle. Strips of portal vein were Na+ loaded for 3 hours at 37 degrees C in a K+-free 1 mM ouabain solution, after which rapid Na+ efflux was induced by washing with a Na+-free K+-Li+ solution (1 mM ouabain). Li+ washing Na+-loaded portal vein produced a large transient contraction accompanied by an increase (over 100-fold) in mitochondrial Ca2+ and also significant (p less than 0.05) increases in phosphorus and Mg2+. The Ca2+ loading of the mitochondria was reversed during prolonged Li+ wash, and by 2 hours, mitochondrial Ca2+, Mg2+, and phosphorus had returned to control levels. The maximal contractile response to stimulation remained normal, demonstrating that pathologic Ca2+ loading of mitochondria is reversible in situ and compatible with normal maximal force developed by the smooth muscle. Mitochondrial Ca2+ and phosphorus uptake were reduced but still significant when the Li+ wash contained 0.2 mM Ca2+ or when ouabain was omitted. The fact that mitochondrial Ca2+ loading accompanied submaximal contractions during 0.2 mM Ca2+-Li wash suggests "supranormal" affinity of mitochondria for Ca2+ and may be due, in part, to reverse operation of the mitochondrial Na+-Ca2+ exchanger. Mitochondrial Ca2+, Mg2+, and phosphorus uptake were eliminated when the Li+ wash was performed at 2 degrees C or when the wash contained no Ca2+.(ABSTRACT TRUNCATED AT 250 WORDS)     

Electrophysiologic properties of a new antiarrhythmic drug--tocainide

Tocainide (Astra W36095) is an orally active antiarrhythmic drug that is structurally related to lidocaine. This paper describes in vivo and in vitro studies undertaken to assay the electrophysiologic properties of this compound. Ouabain-induced ventricular ectopic activity was abolished by this drug in both isolated canine Purkinje fibers and in intact dogs. Similarly, the ventricular ectopic activity that occurs 24 hours after the two stage Harris coronary arterial ligation procedure also was abolished by tocainide. The amount of current necessary to evoke an action potential by intracell stimulation during diastole was Increased in direct relation to drug concentration. Similarly, in dogs anesthetized with pentobarbital sodium, tocainide increased the ventricular fibrillation threshold up to 150 percent above control values during normal supraventricular rhythm and up to 285 percent after premature ventricular beats. The transmembrane action potential duration and effective refractory period of isolated canine Purkinje fibers were shortened by tocainide. Tocainide depressed atrioventricular nodal conduction in anesthetized dogs, an effect that was magnified in the presence of premature atrial beats but had no effect on His, Purkinje or ventricular muscle conduction. These results Indicate that tocainide is a potentially effective antiarrhythmic agent and deserves further investigation in patients.     

The action of theophylline on potassium uptake in cardiac purkinje fibers

The effect of theophylline on potassium uptake of cardiac Purkinje fibers was studied under different conditions. The following results were obtained: (1) theophylline decreases K uptake and enhances Purkinje fiber automaticity; (2) norepinephrine and high [Ca]0 counteract the decrease in K uptake induced by theophylline; (3) adrenergic receptor blockers do not modify the effect of theophylline on K uptake; (4) norepinephrine increases K uptake and this effect is potentiated by theophylline and is little affected by imidazole and acetylcholine. It is concluded that theophylline may depress K uptake by releasing intracellular calcium and thereby inhibiting the sodium-potassium pump. However, theophylline potentiates the effect of norepinephrine on K uptake probably by enhancing the intracellular levels of cyclic AMP.     

K+ efflux in deoxygenated sickle cells in the presence or absence of DIOA, a specific inhibitor of the [K+, Cl-] cotransport system

The ouabain bumetanide resistant (OBR) K+ efflux was investigated in deoxygenated sickle cells in comparison to oxygenated ones, by using a specific inhibitor of the [K+, Cl-] co-transport system, [(DihydroIndenyl)Oxy] Alkanoic acid (DIOA). A DIOA sensitive and a DIOA resistant K+ efflux were measured in deoxygenated sickle cells. The DIOA sensitive K+ efflux shared the properties of the [K+, Cl-] co-transport system, being stimulated by decreased pH and hypoosmolarity. This DIOA sensitive K+ efflux represented 70% of the total K+ efflux at pH 7.0 and at low pO2 (10-15 mmHg). Thus, a small reduction in Ph effectively stimulated the [K+, Cl-] co-transport system in deoxygenated condition, and this may contribute significantly to the sickle cell dehydration. We conclude that at pH lower than 7.4, the [K+, Cl-] co-transport system is permanently activated in sickle cells and leads to sickle cell dehydration in both oxygenated and deoxygenated conditions.     

Reversibility of Electrophysiologic Abnormalities of Subendocardial Purkinje Fibers Induced by Ischemia

Subendocardial Purkinje Fibers. Introduction: During the subacute phase of infarction in the canine heart, the Subendocardial Purkinje fibers subtended by the infarct show depolarization greater than can be accounted for by the decrease in (K), and generate abnormal action potentials and spontaneous rhythms due to abnormal automaticity. We have used pinacidil to hyperpolarize these fibers and evaluate the extent to which an increase in resting potential can normalize action potential generation. Methods and Results: Twenty-four hours after two-stage ligation of the canine left anterior descending coronary artery, preparations of Subendocardial Purkinje fibers were studied in vitro by recording transmembrane potentials through standard microelectrodes and exposing the preparation to pinacidil and increases in (K⁺)₀. Pinacidil increased resting potential to the estimated value of E_K, abolished the abnormal automaticity, and restored action potentials of normal amplitude with normal values of V_max. This effect often persisted after washout of pinucidil. Elevation of (K⁺⁾, from 4.0 to 20.0 mM slightly increased maximum diastolic potential, suggesting (hat the excess (over the change in E_K) depolarization was caused by a decrease in g_Kl. Conclusion: The ventricular arrhythmias seen during the subacute stage of infarction probably are caused by abnormal automaticity. Our findings support the conclusion that this abnormal automaticity arises in partially depolarized suhendocardial Purkinje fibers. This loss of resting potential is due in large part to a decrease in g_K1. Restoration of resting potential to the value of E _k permits the Purkinje fibers to develop essentially normal action potentials. An agent capable of reversing the partial block of 1_K1, thus might be an effective drug for some types of arrhythmias.     

Effects of forskolin on intracellular sodium activity in resting and stimulated cardiac Purkinje fibers from sheep

In the present investigation, the effects of forskolin on intracellular sodium activity were studied in quiescent and electrically stimulated cardiac Purkinje fibers from sheep using Na+-sensitive microelectrodes. Also assessed, were the effects of this promoter of cytosolic cAMP production on resting membrane potential, action potential and twitch tension. In the quiescent fibers, forskolin (12 microM) caused intracellular sodium activity to decrease in the face of cellular depolarization. This cellular depolarization was occasionally accompanied by spontaneous firing of action potentials. In the stimulated fibers, forskolin (10 microM) also caused intracellular sodium activity to decrease. Moreover, it caused a marked acceleration of phase 4 pacemaker depolarization, an elevation of the plateau of the action potential and an increase in twitch tension. When the Na+ pump was inhibited by either strophanthidin (1 microM) or by 0 mM extracellular K+, forskolin had no effect on intracellular sodium activity. In summary, the results of the present study indicate that forskolin, presumably by increasing intracellular cAMP, causes the following to occur in cardiac Purkinje fibers from sheep: (a) a decrease in intracellular sodium activity when the Na+ pump is functioning normally; (b) a promotion of membrane depolarization in quiescent fibers; (c) an increase in the steepness of the pacemaker potential in electrically stimulated fibers, and (d) an increase in the force of contraction. Therefore, forskolin will be a useful tool for investigating the role of cAMP in physiological function of cardiac cells.     

Electrophysiological observations on the digitalis-potassium interaction in canine Purkinje fibers.

We studied the effects of elevating potassium concentration on the membrane potential of Purkinje cells exposed to toxic concentrations of acetylstrophanthidin or ouabain. Conventional intracellular microelectrode techniques were employed. Rapid elevation of [K+]o from 2.7 to 5.4 mEq/liter resulted in an initial increase (more negative) in membrane potential of cells demonstrating ouabain-induced phase 4 depolarization. The increase in maximal diastolic potential occurred initially without suppression of phase 4 depolarization. In cells rendered inexcitable by ouabain or acetylstrophanthidin, elevation of [K+]o consistently increased membrane potential and restored excitability. In four experiments automaticity was initiated within 2 minutes after the increase in [K]o. Although automaticity reappeared, as maximal diastolic potential increased, the automatic rate slowed and then pacemaker activity was suppressed. Studies with 3H-ouabain showed that the increase in membrane potential paralleled K+-induced release of 3H-ouabain from Purkinje cells. These studies suggest that elevation of [K+]o reverses digitalis toxic manifestations in canine Purkinje fibers by causing release of cardiac glycosides bound to the membrane. The observed increase in membrane potential of ouabain-treated Purkinje fibers that occurred after [K+]o elevation was considered to be mediated in part by restoration of the Na pump and by electrogenic pumping.     

Electrotonic Inhibition and Active Facilitation of Excitability in Ventricular Muscle

Inhibition and Facilitation in Cardiac Muscle. Introduction: The effects of subthreshold electrical pulses on the response to subsequent stimulation have been described previously in experimental animal studies as well as in the human heart. In addition, previous studies in cardiac Purkinje fibers have shown that diastolic excitability may decrease after activity (active inhibition) and, to a lesser extent, following subthreshold responses (electrotonic inhibition). However, such dynamic changes in excitability have not been explored in isolated ventricular muscle, and it is uncertain whether similar phenomena may play any role in the activation pal-terns associated with propagation abnormalities in the myocardium. Methods and Results: Experiments were performed in isolated sheep Purkinje fibers and papillary muscles, and in enzymatically dissociated guinea pig ventricular myocytes. In all types of preparations introduction of a conditioning subthreshold pulse between two subthreshold pulses was followed by a transient decay in excitability (electrotonic inhibition). The degree of inhibition was directly related to the amplitude and duration of the conditioning pulse and inversely related to the postconditioning interval. Yet, inhibition could be demonstrated long after (> 1 sec) the end of the conditioning pulse. Electrotonic inhibition was found at all diastolic intervals and did not depend on the presence of a previous action potential. In Purkinje fibers, conditioning action potentials led to active inhibition of subsequent responses. In contrast, in muscle cells, such action potentials had a facilitating effect (active facilitation). Electrotonic inhibition and active facilitation were observed in both sheep ventricular muscle and guinea pig ventricular myocytes. Accordingly, during repetitive stimulation with pulses of barely threshold intensity, we observed: (1) bistability (i.e., with the same stimulating parameters, stimulus: response patterns were either 1:1 or 1:0, depending on previous history), and (2) abrupt transitions between 1:1 and 1:0 (absence of intermediate wenckebach-like patterns). Simulations utilizing an ionic model of cardiac myocytes support the hypothesis that electrotonic inhibition in well-polarized ventricular muscle is the result of partial activation of I_k following subthreshold pulses. On the other hand, active facilitation may be the result of an activity-induced decrease in the conductance of I_K1. Conclusion: Diastolic excitability of well-polarized ventricular myocardium may be transiently depressed following local responses and transiently enhanced following action potentials. On the other hand, diastolic excitability decreases during quiescence. Active facilitation and electrotonic inhibition may have an important role in determining the dynamics of excitation of the myocardium in the presence of propagation abnormalities.     

Role of the conduction system in the endocardial excitation spread in the right ventricle

The role of the Purkinje network in the excitation sequence in the endocardial surface of the right ventricle was studied using an isolated perfused canine preparation. The preparation was electrically stimulated at the proximal right bundle branch, and the activation time was mapped using contiguous bipolar electrodes or a microelectrode, or both. The earliest activation of muscle was observed at the junction between the ventricular septum and the free wall in front of the anterior papillary muscle. After the initial activation, the spread of ventricular muscle excitation in the free wall was essentially radial at a mean (± standard deviation) conduction velocity of 1.67 ± 0.20 m/s. The activation of muscle in this area was almost always preceded by the activation of Purkinje fibers by 2 to 6 ms. Thus the Purkinje system was considered indispensable for the excitation spread in this area. In the lower third of the septum, the excitation sequence was essentially similar to that of the free wall, indicating a possible contribution of the fast conducting Purkinje system. In contrast, in the upper two thirds of the septum the activation of ventricular muscle spread from apex to base with a significantly delayed conduction velocity of 0.41 ± 0.88 m/s, and it was not preceded by activation of Purkinje fibers, thus indicating the lack of involvement of the Purkinje system.Functional distribution of the anterior, posterior and lateral branches of the right bundle to the excitation spread of the right ventricular endocardial muscle was confirmed by the selective transection of one of these special conducting fibers, which suggested the etiologic significance of injury to each branch of the right bundle as a cause of various electrocardiographic patterns of incomplete right bundle branch block.