Faculty Bibliography

STUDY OBJECTIVE/OBJECTIVE:ATXII is a polypeptide toxin isolated from the sea anemone, Anemonia sulcata, known to delay sodium inactivation markedly and to induce early afterdepolarisations. The aim was to investigate the mechanism of its action. DESIGN AND MATERIALS/METHODS:The mechanism of ATXII induced early afterdepolarisations was investigated in vitro in canine endocardial preparations using standard microelectrode techniques. MEASUREMENTS AND MAIN RESULTS/RESULTS:ATXII (2 x 10(-7) M) induced cycle length dependent prolongation of plateau, more marked in Purkinje than in muscle fibres, and early afterdepolarisations in Purkinje fibres only. The calcium channel antagonists verapamil (10(-6) M, 10(-5) M) and cobalt (2-4 mM), and drugs that block calcium release from the sarcoplasmic reticulum, ryanodine (10(-6) M, 10(-5) M) and caffeine (10 mM), did not antagonise the ATXII effects. However, tetrodotoxin (5 x 10(-6) M) and lignocaine (4 x 10(-5) M) shortened the action potential and suppressed early afterdepolarisations. The effects of lignocaine were seen at concentrations that did not significantly affect Vmax. CONCLUSIONS:ATXII induced early after depolarisations are due to the effects of ATXII on Na+ entry, probably via a slowly inactivated Na+ channel population. Calcium entry through the sarcolemmal Ca2+ channels and cyclic Ca2+ release from the sarcoplasmic reticulum are not required for the genesis of early afterdepolarisations in this model.

The reality of sinoatrial Wenckebach periods (WP) has been suggested, but not proven, in the literature. We report experimental and clinical data showing WP in sinoatrial blocks. Experimental sinoatrial blocks were induced by superfusion of bepridil (10(-5) M) in 15 preparations of isolated rabbit right atria. Different types of block were observed, including Blumberger I block, i.e., sinoatrial WP. The recordings showed that the typical pattern of Blumberger type IA block and sinoatrial WP may be due to transient acceleration of the sinus rate, without change in the increment. We also observed sinoatrial WP in a 72-year-old patient on direct recordings of the sinus node (SN) electrical activity. In this case, transient acceleration of the sinus rate also seemed to be involved in the genesis of sinoatrial WP. Analysis of these clinical and experimental data showed similarities that may explain the mechanism of the WP. Usually type IA Blumberger block is said to involve a decrease in the sinoatrial increment to explain the atrial sequence (decrease in PP interval followed by a pause shorter than twice the value of the preceding cycle, then a cycle longer than the one preceding the pause). In fact, this pattern can be observed when the acceleration of the higher structure, the SN, induces block within the lower structure, i.e., the atrium.

Alpha 1-adrenoceptor agonists were shown to induce delayed afterdepolarizations (DADs) and triggered activity in the presence of elevated extracellular Ca2+. We investigated the effects of alpha 1-adrenoceptor stimulation on DADs and triggered activity in canine Purkinje fibers that survived 1-day of myocardial infarction. Endocardial preparations were studied using standard microelectrode techniques. In quiescent preparations showing no DADs and in presence of propranolol (2 x 10(-7) M), phenylephrine (10(-6) M), an alpha 1-adrenoceptor agonist induced DADs (n = 6) and differentially induced triggered activity in ischemic but not in normal Purkinje fibers (n = 4). In 8 preparations that showed subthreshold DADs, phenylephrine increased the DAD amplitude from 4.0 +/- 2.5 mV to 8.0 +/- 3.3 mV (P less than 0.03) and from 3.2 +/- 1.5 mV to 6.5 +/- 3.7 mV (P less than 0.05) at paced cycle lengths of 800 and 400 ms, respectively. Phenylephrine caused subthreshold DADs to reach threshold and result in triggered activity (n = 6). The effects of phenylephrine were abolished by 10(-6) M prazosin, an alpha 1-adrenoceptor blocker. Our results suggest that alpha 1-adrenoceptor stimulation regulates DADs and triggered activity seen in subendocardial Purkinje fibers surviving 1 day of myocardial infarction and may contribute to the spontaneous ventricular tachycardia seen in vivo at this stage.

The diterpene forskolin is widely known for its ability to directly activate adenylyl cyclase and consequently increase intracellular cAMP. In cardiac cells, one result is a cAMP-mediated increase in the L-type Ca2(+)-channel current (ICa). However, forskolin was also shown recently to affect a number of ionic channels in noncardiac cells by mechanisms that do not involve activation of adenylyl cyclase. The present study reveals such an effect of forskolin on cardiac Ca2+ channels. Indeed, under appropriate conditions, forskolin was found to cause an inhibition of ICa. Although the stimulation of adenylyl cyclase and ICa requires micromolar concentrations of forskolin, the inhibitory effect of forskolin was observed in the nanomolar range of concentrations, i.e., 2-3 orders of magnitude lower. This high affinity forskolin inhibition of ICa was observed when ICa was previously enhanced via a cAMP-dependent pathway, but not when ICa was at its basal level or when the current was elevated by the dihydropyridine Bay K 8644. The inhibitory effect occurred at a site of action remote from adenylyl cyclase, because forskolin similarly inhibited ICa that had been previously elevated by isoprenaline (a beta-adrenergic agonist) or directly by intracellular perfusion with cAMP. Under these conditions, forskolin was inhibitory when applied to either side of the cell membrane, but only in its lipid-soluble form. The inhibitory effect of forskolin appeared to be independent of membrane potential and was not accompanied by a change in the time constants of ICa activation and inactivation. This may indicate that forskolin mainly reduces the number of functional Ca2+ channels without changing the gating of individual channels. However, the reduction in ICa amplitude was not equally distributed among the different exponential components that constitute ICa, which suggests that forskolin also modifies the resting state of the channels. This novel high affinity forskolin inhibition of ICa may take place at some step in the pathway between cAMP and Ca2+ channel phosphorylation and/or at Ca2+ channels only after they have been phosphorylated.

New information about the pathophysiology of the sinus node and sino-atrial conduction has been published in the last few years. The sinus node consists of cells separated by a network of collagen fibres. This anatomical disparity explains the different electrophysiological characteristics of the node; the morphology of cellular action potentials depends on the site of recording. The dominant and most automatic pacemaker cells are situated in the cephalic region and the latent pacemaker cells in the caudal region. However, synchronisation of these different cellular activities is possible and results in a coherent signal. This complex synchronisation has been the object of several recent papers. The phenomenon of intrasinusal pacemaker shift and the stimuli which induce it have been studied in depth. In general, positive chronotropic stimuli tend to shift the dominant pacemaker towards the cephalic part and negative chronotropic stimuli towards the caudal part of the node. It is possible to assess pacemaker shift clinically and this phenomenon must be taken into consideration when studying sinus node function. Intercellular conduction and especially electrotonic conduction does not play a role in the genesis of the flux, which represents spontaneous cellular automatism, but in its mode of expression, that is to say the sinus rhythm. The pathophysiology of sinoatrial block is complex because it may be situated within and/or around the sinus node. The extrinsic or intrinsic mechanisms of these blocks may be interrelated. Variations in sinus rhythm must be taken into account in the genesis of sinoatrial block; an acceleration in rhythm may block conduction in the perisinusal region. Finally, our knowledge of the ionic fluxes underlying sinus automatism has also improved with individualization of the pacemaker current (if).(ABSTRACT TRUNCATED AT 250 WORDS)

The increase in extracellular potassium [K+]o levels during the early phase of myocardial ischemia may result in part from activation of adenosine triphosphate-sensitive K+ channels. Glyburide, a second-generation hypoglycemic sulfonylurea, is a potent blocker of these channels. We studied the effects of glyburide on [K+]o and on intramyocardial conduction delay during a 10-minute occlusion of the left anterior descending artery in the dog. K(+)-sensitive electrodes and bipolar plunge electrodes were introduced to record, respectively, [K+]o and local electrograms from close sites in midmyocardial regions in normal, border, and ischemic zones. Recordings were obtained before (control ischemia [CI]) and 20 minutes after intravenous administration of 0.15 mg/kg of glyburide (glyburide plus ischemia [G + I]). During G + I the extent of the increase in [K+]o was less compared to that during CI, and the difference was statistically significant during the first 7 minutes of ischemia in the ischemic zone and during the first 4 minutes of ischemia in the border zone. On the other hand, the degree of local intramyocardial conduction delay was significantly reduced during G + I compared to CI during the entire 10 minutes of ischemia in both the ischemic and border zones. In summary, our results have shown that glyburide significantly reduced the rise of [K+]o and intramyocardial delay during the early phase of acute ischemia and could thus attenuate the electrophysiologic consequences of ischemia that underlie the initial phase of malignant tachyarrhythmias. Although the effects of glyburide may result in part from a direct action of the drug on cardiac adenosine triphosphate-sensitive K+ channels, other metabolic antiischemic effects cannot be ruled out.

Caffeine and ryanodine are known to modulate oscillatory release of Ca2+ from the sarcoplasmic reticulum. The effects of caffeine and ryanodine on delayed afterdepolarizations (DADs) and sustained rhythmic activity in subendocardial Purkinje fibers surviving 1-day-old myocardial infarction in the dog were studied with standard microelectrode techniques. In preparations that showed sustained rhythmic activity, a high concentration of caffeine (10 mM) and ryanodine (10(-7) and 10(-6) M) slowed and terminated the sustained rhythmic activity and markedly suppressed DADs. An increase in the temperature of the tissue bath from 37 degrees to 39 degrees C did not change these results. In quiescent normal and infarcted preparations, a low concentration of caffeine (0.5 mM) differentially induced DADs in ischemic but not in normal Purkinje fibers, increased the amplitude of existing DADs, and brought subthreshold DADs to threshold potential that caused triggered activity. Our results are consistent with the hypothesis that triggered activity arising from DADs characterizes the sustained rhythmic activity in endocardial preparations 1 day after infarction and indicate an important role for the sarcoplasmic reticulum in the genesis of DADs and triggered activity in this model.

Factors that facilitate the occurrence of cholinergic atrial arrhythmias were studied on 16 rabbit biatrial preparations in spontaneous rhythm. Sinus cycle length and characteristics of the atrial action potential were measured by the microelectrode technique in the basal state, then in the presence of acetylcholine at a concentration of 1.4 x 10(-5) M. Induction of arrhythmia was attempted by programmed stimulation, using an increasing number of extrastimuli. In the presence of acetylcholine the sinus cycle length increased by 106 +/- 63 p. 100 (p less than 0.0001) and the action potential duration, measured at 90 p. 100 repolarization (APD90) decreased from 60 +/- 15 ms to 40 +/- 11 ms (p less than 0.001). Reentrant activities, which had not been found in the basal state, were induced in 5 preparations. Under acetylcholine the sinus cycle of inducible preparations was shorter than that of non inducible preparations (663 +/- 272 ms vs 1218 +/- 531 ms, p less than 0.05). The percentage of sinus cycle lengthening was significantly smaller in inducible preparations (54 +/- 31 p. 100 vs 129 +/- 60 p. 100, p less than 0.05). Although sinus cycle lengthening was different in the two types of preparations, the APD90 was shortened in the same proportions. The vulnerability of the preparations seemed to depend mainly on a frequency effect. Vagal atrial arrhythmias occurred with a relatively small reduction in sinus rhythm. It is probable that an overpotent vagal effect is less arrhythmogenic because of its more homogeneous action on tissues.

In order to determine the respective roles of catheter (Ct) physical properties and of energy levels in myocardial effects of fulguration, we delivered an electrical shock between the tip electrode of a Ct placed at the apex of the right ventricle and a large cutaneous cathodal electrode in 12 dogs. Two energy levels were used: Group A = 25 J (n = 6) and group B = 100 J (n = 6), and three Cts were studied. These Cts had different resistances (R) and active surface electrodes (S): Ct 1 (R = 0.3 omega, S = 12 mm2), Ct 2 (R = 0.3 omega, S = 2 mm2), Ct 3 (R = 2 omega, S = 13 mm2). Complex ventricular arrhythmias were observed in 5/6 cases at 100 J but only in 1/6 cases at 25 J and were independent of the Ct type. Following the shock, the effective ventricular refractory period (S1 S1 = 300 msec) increased significantly only at 100 J (11%, p = 0.03). Anatomical lesions were wider (10.6 vs. 5.2 mm, p less than 0.05) and deeper (100 vs. 55%, p less than 0.05) in the 100 J group. In contrast, there was no significant difference in the electrophysiological and anatomical changes between the three Cts. In conclusion, arrhythmogenic adverse effects of ventricular Ct fulguration are related to the delivered energy; on the contrary, they seem only slightly dependent on Ct physical properties at these energy levels; a 2 J/kg shock is not only effective but also seems to be safe.

Spatial inhomogeneity of refractory periods, as measured during clinical electrophysiological studies, is a known predisposing factor of arrhythmia. We studied effective refractory periods (ERP) and action potential duration (ADP90) on isolated human atrium. Twelve samples of right atrium obtained during cardiac surgery from patients with (n = 6) and without (n = 6) atrial fibrillation (AF) were studied by microelectrode technique. For each preparation, ERP were measured at basic cycle lengths (BCL) of 1,600, 1,200, 800, and 400 msec in five different cells located around (0.8 mm) the stimulating electrode. Dispersion of ERP was significantly greater in the AF group (96.7 +/- 9 versus 70.9 +/- 9 msec, p = 0.01). In the non-AF group, we observed a positive linear correlation between ERP and BCL (r = 0.86) ADP90 and BCL (r = 0.93). On the contrary, in the AF group this correlation was absent between ERP and BCL (r = 0.28), poor between ADP90 and BCL (r = 0.62). These results suggest that nonhomogeneous recovery of excitability (dispersion and poor adaptation) may be an important factor of arrhythmia. This inhomogeneity is present at the cellular level as well as in the entire heart.