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G Vassort

Publications and source records attributed to G Vassort.

At least 19 recordsLinked to original sources
PubMed1992

Protein kinase C enhances myosin light-chain kinase effects on force development and ATPase activity in rat single skinned cardiac cells.

Many neurohormones alter the force of cardiac contraction by variations in the intracellular Ca2+ concentration. alpha 1-Adrenergic and muscarinic stimulations, rather, modify the sensitivity of contractile proteins to Ca(2+)-calmodulin-myosin light-chain kinase (MLCK) complex induces a large increase in Ca2+ sensitivity (0.14 pCa unit) of these easily accessible myofilaments. This increase is further enhanced by up to 0.19 pCa unit when protein kinase C (PKC) is added together with MLCK. Similarly, the Ca2+ ATPase activity of skinned cells in suspension is increased in the presence of MLCK and further in the presence of both kinases. 32P-labelling and SDS/PAGE show that these changes are associated with light-chain 2 (LC2) phosphorylation together with phosphorylation of troponin I and troponin T when PKC is added. Although to a smaller extent than in smooth muscle, phosphorylation of cardiac myosin LC2 may be involved in the modulation of heart contractility.

Adenosine Triphosphatases↗
PubMed1992

Phenylephrine and ATP enhance an amiloride insensitive bicarbonate-dependent alkalinizing mechanism in rat single cardiomyocytes.

To expel the excess protons generated during a cellular acidification and to fully recover basal intracellular pH (pHi), cardiac cells rely on the amiloride-sensitive Na/H antiport. We report that rat single ventricular cardiomyocytes, loaded with the fluorescent pH indicator Snarf-1 and treated with inhibitors of the Na/H antiport, amiloride or its analogues, partially restored their pHi through a bicarbonate-dependent mechanism following an acidosis (imposed by the ammonia-pulse technique). In the presence of ethylisopropylamiloride (10 microM) or amiloride (1 mM) and 25 mM bicarbonate in the extracellular solution, the average time that cells needed to recover half of their pHi, following the removal of 20 mM NH4Cl, was 3.4 min, while the rate of proton efflux was calculated to be 2.0 mM/min. The stilbene derivative, 4-4'-di-isothiocyanostilbene-2,2'-disulphonate (DIDS 200 microM), a known blocker of anion transporters, inhibited this recovery. Both phenylephrine (100 microM, 3 microM propranolol present), an alpha 1-adrenoceptor agonist, and ATP (10 microM), a purinergic agonist, significantly enhanced the rate of proton efflux that was due to this HCO3-dependent alkalinizing mechanism. Phenylephrine and ATP also shortened by three-fold the time that a myocyte needed to recover half of its initial pHi. This bicarbonate-dependent alkalinizing mechanism could provide an additional means by which cardiac cells recover their pHi from acidosis, especially under conditions in which the Na/H antiport is inhibited. Furthermore, catecholamines and ATP, which are released under various pathophysiological conditions often associated with intracellular acidosis, could play an important role in the modulation of pHi under these conditions.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1992

Properties of the low threshold Ca current in single frog atrial cardiomyocytes. A comparison with the high threshold Ca current.

The properties of the low threshold Ca current (ICaT) in bullfrog (Rana catesbeiana) isolated atrial cardiomyocytes were studied using the whole-cell recording patch-clamp technique and compared with those of the high threshold Ca current (ICaL). In 91% of atrial cells we observed both ICaT and ICaL when collagenase and trypsin were used to dissociate the cells. But when pronase was used, only 30% of the cells exhibited ICaT. ICaT was never found in ventricular cells. ICaT could be investigated more easily when ICaL was inhibited by Cd ions (50 microM). Its kinetics were unchanged by substituting Ba for Ca, or in the presence of high concentrations of Ba. Both ICaT and ICaL exhibited reduced inactivation after high depolarizing prepulses. ICaT was found to be sensitive to dihydropyridines: 1 microM nifedipine decreased this current while 1 microM BAY K 8644 increased it; this occurred without significant variations in the steady-state inactivation curve. ICaT was more sensitive than ICaL to alpha 1-adrenergic and P2-purinergic stimulations, while ICaL was more sensitive to beta-adrenergic stimulation. Isoproterenol was still able to increase ICaT in the presence of high intracellular cAMP. Both currents were increased by 1 microM ouabain (although ICaL only transiently) and decreased by 10 microM ouabain. It is concluded that the two types of Ca channels can be observed in bullfrog atrial cells and that they are specifically altered by pharmacological agents and neuromediators. This may have implications for cardiac behavior.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed1992

A Gs protein couples P2-purinergic stimulation to cardiac Ca channels without cyclic AMP production.

P2-purinergic stimulation of the L-type Ca current induced by the external application of 100 microM ATP gamma S was investigated in rat ventricular cardiomyocytes using the whole-cell patch-clamp technique. The purinergic-induced increase in ICa was slow and monophasic and reached a steady state within 3 min. In contrast to beta-adrenergic stimulation, after a brief agonist application the current did not continue to increase on washout; recovery started immediately after agonist removal. The P2-purinergic increase in ICa was significantly less in the presence of GDP beta S, but it occurred much faster and was twice as large when a low dose of GTP gamma S (100 microM) was added to a GTP-containing internal medium. This suggests that the ICa increase was mediated by a G protein. Based on electrophoretic mobility and susceptibility to cholera toxin and anti-G alpha s serum, it is proposed that the G protein involved during purinergic-induced ICa stimulation is an isoform of Gs not coupled to the adenylyl cyclase, since the cyclic AMP level was unaffected. High intracellular GTP gamma S (1 mM) maximally activated ICa so that neither beta-adrenergic nor P2-purinergic agonists further increased ICa. In the absence of GTP and an ATP-regenerating system, GTP gamma S was much more potent in increasing basal ICa and supporting purinergic stimulation. This indicates that a nucleoside diphosphate kinase activity might replenish endogenous GTP; GTP exchange with GTP gamma S on the G protein was promoted by the P2-purinergic stimulation and led to a reversible and reproducible increase in ICa. In the presence of 3 mM internal ATP gamma S, the P2-purinergic stimulation was also reversible and reproducible. Moreover, under these conditions (ATP gamma S or GTP gamma S) the increase in ICa was not maintained during prolonged agonist application. Such an inhibition occurred slowly and irreversibly; it might be related to the threefold increase in cyclic GMP. In conclusion, we propose that extracellular ATP induces both a stimulatory and an inhibitory effect on ICa, probably mediated by subtypes of P2-purinergic receptors. An isoform of the Gs protein is likely to mediate the stimulation.

Adenosine Diphosphate↗
PubMed1992

Alpha 1-adrenergic effects on intracellular pH and calcium and on myofilaments in single rat cardiac cells.

1. The cellular effects of alpha 1-adrenoceptor stimulation by phenylephrine were studied in the presence of propranolol in single cells isolated from the ventricles of rat hearts. 2. Phenylephrine (10-100 microM) induced a biphasic pattern of inotropism in these cells: a transient negative followed by a sustained positive inotropic effect as usually observed in cardiac tissues. 3. In Snarf-1-loaded cells, phenylephrine induced an alkalinization. This effect was reversible on wash-out and inhibited by prazosin, an alpha 1-adrenoceptor antagonist. 4. The alpha 1-adrenoceptor-mediated increase in intracellular pH (pHi) was 0.1 pH unit in HEPES buffer containing 4.4 mM-NaHCO3 and in Krebs buffer containing 25 mM-NaHCO3. 5. The alkalinization was blocked by the Na(+)-H+ antiport blocker, ethylisopropylamiloride (EIPA). 6. The recovery from an acidosis induced by a NH4Cl pre-pulse was accelerated by phenylephrine. The phenylephrine-induced alkalinization was attributed to activation of the Na(+)-H+ antiport. 7. Despite its ability to increase pHi, phenylephrine did not alter Ca2+ current amplitude and kinetics. 8. Ca2+ transients recorded in Indo-1-loaded cells were not augmented by phenylephrine. Diastolic calcium level was decreased. 9. In single skinned cells, the Ca2+ sensitivity of the contractile proteins was increased by a pre-treatment with phenylephrine even when the alpha 1-adrenoceptor-mediated alkalinizing effect had been prevented by EIPA. 10. These results lead us to propose that the alpha 1-adrenergic-induced positive inotropic response of heart muscle could result from an increased sensitivity of the myofilaments to Ca2+ ions. This alpha 1-adrenoceptor-mediated Ca2+ sensitization could result both from an intracellular alkalinization and from a direct effect on contractile proteins.

Amiloride↗
PubMed1992

Prajmalium, an antiarrhythmic with positive inotropic effect: mechanism of action in rabbit single cardiomyocytes.

The propyl derivative of ajmaline, N-n-propylajamaline (prajmalium), is an antiarrhythmic compound that lacks the commonly reported negative inotropic effects of all others under clinical use. The present study was undertaken to establish and understand its effects at the cellular level in mammalian preparations. Electrical and mechanical activities were recorded from right ventricular strips and Na and L-type Ca currents (INa and ICaL, respectively) were recorded with the whole-cell patch-clamp technique in right ventricular myocytes freshly dissociated from rabbit hearts. Prajmalium decreased the maximal rate of depolarization of the action potential in a dose-dependent manner with an EC50 of 3 microM. This effect was use and frequency dependent. Action potential duration was increased by 1 microM prajamalium but decreased on applying higher concentrations. The force of contraction was slightly (15%) increased at 0.1 microM, not affected at all at 1 microM and depressed by 30% at 20 microM. In single cardiomyocytes maintained at negative holding potentials, INa was slightly depressed by prajmalium at 10 nM and reduced by 75% at 10 microM. ICaL was increased by 30 and 20% on applying prajmalium at 1 and 10 microM, respectively; on the other hand, ICaL was reduced by these two concentrations of prajmalium at less negative holding potentials. A higher prajmalium concentration (100 microM) decreased ICaL at all holding potentials studies and this effect was enhanced with depolarization. The increase in ICaL induced by prajmalium (1 microM) was also observed after ICaL had been fully beta-adrenergic and P2-purinergic stimulated by isoproterenol (1 microM) in the presence of IBMX (100 microM) and ATP (10 microM). It is concluded that prajmalium is able to increase ICaL in rabbit ventricular cells in a voltage-dependent manner, an effect that could account in part for the observed lack of negative inotropism of this antiarrhythmic in clinics.

Action Potentials↗
PubMed1991

Extracellular ATP-induced acidification leads to cytosolic calcium transient rise in single rat cardiac myocytes.

The origin of the increase in cytosolic free Ca2+ concentration ([Ca2+]i) induced by extracellular ATP was investigated in single isolated cardiac myocytes loaded with indo-1. The nucleotide added at a concentration of 10 microM triggers a few Ca2+ spikes, followed by a cluster of Ca2+ oscillations, increasing [Ca2+]i to around 200 nM from a basal value of 70 nM. Neither caffeine nor ryanodine affects the magnitude of the Ca2+ transient, but both shorten it by preventing the Ca2+ oscillations. This indicates that the latter must be related to the release of Ca2+ from the sarcoplasmic reticulum. Since ATP also induces cell depolarization (as shown by experiments using the potential sensitive dye bis-oxonol), the initial Ca2+ spikes were attributed to the opening of voltage-dependent Ca2+ channels. A small Ca2+ transient still remains under experimental conditions designed to prevent Ca2+ influx from external medium (low-Ca2+ high-Mg2+ medium containing La3+) and after depletion of the sarcoplasmic-reticulum Ca2+ load with caffeine. Under these conditions, when this Ca2+ transient was buffered by 1,2-bis-(O-aminophenoxy)ethane-NNN'N'-tetra-acetic acid, ATP was unable to trigger the initial Ca2+ spikes. These results indicate that ATP mobilizes Ca2+ ions from an intracellular pool other than the sarcoplasmic reticulum and that this Ca2+ release is responsible for the depolarization. The effects of ATP on [Ca2+]i share the same characteristics as the acidification simultaneously induced by the nucleotide (as shown by experiments using the pH-sensitive probe snarf-1). These ionic variations are highly specific to ATP and its hydrolysis-resistant analogues. They both require the presence of Mg2+ and Cl- ions in the extracellular medium, and they are prevented by pretreatment of the cells with 4,4'-di-isothiocyanostilbene or probenecid. These results suggest that: (1) the ATP-induced acidification leads to displacement of Ca2+ ions from or close to the internal face of sarcolemma; (2) the Ca2+ ions activate a non-specific membrane conductance responsible for the depolarization of the cells; (3) the depolarization leads to a Ca2+ influx, owing to the opening of the voltage-dependent Ca2+ channels; (4) this increase in Ca2+ triggers the release of Ca2+ from the sarcoplasmic reticulum, which is facilitated by the increase in inositol trisphosphate following P2-purinergic stimulation.

Adenosine Triphosphate↗
PubMed1991

Dual action of prajmalium on the Ca currents in frog isolated cardiomyocytes.

The effects of N-n-propylajmaline (prajmalium) on the Na and Ca currents of single frog atrial and ventricular cells were studied by means of the whole-cell patch-clamp technique. Prajmalium (10(-9) to 10(-6) M) depressed the Na current (INa) in a dose- and use-dependent manner. In the same range of concentrations, prajmalium induced a dual effect on the high (ICaL) and low (ICaT) threshold Ca currents (the latter being only present in atrial cells). At a low concentration (10(-9) M), prajmalium increased both Ca currents while at high concentrations (10(-6) M) it depressed them. Prajmalium appeared very potent on ICaT although this current is generally reported to be barely sensitive to agonists and drugs. The action of the drug was also accompanied by a shortening in the half-time of inactivation of the Ca currents and a slight hyperpolarizing shift of their availability curves. The increase in ICaL by prajmalium was not prevented by prazosin (10(-7) M) nor by propranolol (10(-6) M), and it was also observed after ICaL had been fully stimulated by isoproterenol (10(-7)M). Nifedipine (10(-6) M), however, was able to prevent or block the prajmalium-induced increase in ICaL. Some similarities between the actions of prajmalium and dihydropyridine agonists on Ca currents are discussed.

Animals↗
PubMed1991

Extracellular MgATP activates the Cl-/HCO3- exchanger in single rat cardiac cells.

1. The effect of extracellular MgATP on cytosolic pH (pHi) was investigated in single rat cardiac cells loaded with the pH-sensitive probe Snarf-1. 2. Basal pHi in HEPES-buffered solution (containing 4.4 mM-NaHCO3) was 7.08. MgATP induced a transient acidification followed by an alkalinization. The latter is prevented by ethylisopropylamiloride (EIPA) and has been attributed to the activation of the Na+/H+ antiport. The MgATP-induced acidification reached a maximal value of 0.42 +/- 0.03 pH units (U pH). It was concentration dependent with a K0.5 of 2.6 microM-MgATP. This acidification was also observed with the same magnitude in the presence of the more physiological Krebs-bicarbonate buffer but was greatly reduced in nominally HCO3-free HEPES. 3. The MgATP-induced acidification was prevented by 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS), probenecid and ethacrynic acid but not by bumetanide. It was dependent upon the external chloride concentration. The K0.5[Cl-] was 9 mM and the maximal acidification required 60 mM-Cl-. 4. MgATP accelerated the recovery from an alkalinization triggered by a pulse of NH4Cl. The nucleotide also facilitated the efflux of HCO3- when the cell was switched from a Krebs-bicarbonate buffer gassed with 5% CO2 to an HEPES buffer. 5. The acidification was only evoked by MgATP and its poorly hydrolysable analogues but not by the other nucleotides (ADP, GTP (guanosine triphosphate), CTP (cytidine triphosphate) UTP (urodine triphosphate), ITP (inositol triphosphate) nor by adenosine. It required the presence of Mg2+ ions. 6. These results provide evidence that MgATP activates the Cl-/HCO3- exchanger and that this activation accounts for the acidification. Such an activation could not be related to the P1- or the P2-purinergic receptors since it requires triphosphate adenylic compounds and Mg2+ ions. This leads us to suggest the existence of a putative P3-type of purinergic receptor.

Adenosine Triphosphate↗
PubMed1990

Effects of purinergic stimulation on the Ca current in single frog cardiac cells.

Ca current (ICa) was measured by whole-cell voltage clamp in single cells isolated from frog ventricle, in which the Na current was inhibited by tetrodotoxin (0.3 microM) and K currents were blocked by substituting K with 120 mM intracellular and 20 mM extracellular Cs. The influence of stimulation by ATP (0.1-100 microM) was assessed in the presence of propranolol (1 microM) or pindolol (0.1 microM), prazozin (0.1 microM) and atropine (10 microM). ATP, in the micromolar range, had two types of effect. Like other P1-purinoagonists, it antagonized the increase in ICa elicited by beta-adrenostimulation. When added alone, 1 microM ATP could increase ICa up to twofold. An increase in ICa was also observed even after it had been maximally enhanced by intracellularly applied cAMP (50 microM). Voltage dependence and kinetics of ICa were not affected. These effects were considered to be related to P2-purinoceptor activation. At higher ATP concentrations the increase in ICa was less; at 100 microM, ATP reduced ICa. The ATP-induced increase in ICa was prevented by internal perfusion of the cells with GDP [beta-S] or neomycin, respectively, to block signal transduction to phospholipase C or its phosphodiesterase activity on the polyphosphoinositides. We conclude that P2-purinoceptor stimulation increases the Ca current in frog ventricular cells by a pathway that might involve phosphoinositide turnover.

Adenosine Triphosphate↗
PubMed1990

Mechanism of extracellular ATP-induced depolarization in rat isolated ventricular cardiomyocytes.

Adenosine triphosphate (ATP) is released during neural stimulation and cardiac hypoxia and several mechanisms of its action have been reported in different tissues. ATP stimulates P1 and P2 purinergic receptors; it also activates receptor-operated channels and increases membrane permeability to small ions. In single rat ventricular cells under whole-cell patch-clamp, a stepwise application of ATP in the micromolar range affects the resting potential and membrane currents through an entirely novel mechanism of action which involves several steps. Extracellular ATP induces an inward current and depolarization of the cell, leading to automaticity. The inward current is non-specific for cations, its reversal potential is around -5 mV. The conductance change evoked by ATP is suppressed by 4,4-diisothiocyanostilbene 2,2-disulphonic acid (DIDS) and low-chloride media and is prolonged by adding intracellular bicarbonate. These effects are specific for ATP in the presence of magnesium and are not evoked by a non-hydrolysable analogue of ATP or in the presence of vanadate. Other nucleotides are ineffective. We propose that ATP hydrolysis activates the chloride/bicarbonate (Cl-/HCO3-) exchanger. The induced local acidification could then increase intracellular free calcium and as a consequence, increases the sarcolemmal conductance. Thus, a sudden release of ATP in pathological conditions would induce a depolarization which could generate ventricular arrhythmias.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1990

Inhibition by alcohols, halothane and chloroform of the Ca current in single frog ventricular cells.

Changes in the Ca current, ICa, evoked by ethanol, long chain alcohols (hexanol, octanol and dodecanol) and two halogenated anesthetics (chloroform and halothane) were studied in single isolated frog ventricular cells using the whole cell patch clamp technique. Each of these general anesthetics induced a concentration-dependent decrease in ICa which was usually followed by a partial recovery during long application (5 min). However, an increase in ICa was observed with low ethanol concentrations. The potency of the alcohols was related to their oil-water partition coefficient in that the longer the carbon chain the greater the reduction in ICa. For example, to reduce ICa to half its control amplitude required 300 mM ethanol but only 0.3 microns dodecanol. The kinetics of ICa were also altered by exposure to the alcohols. The halogenated derivatives also decreased ICa and altered the kinetics of ICa, but in a different way to the alcohols. The complex and different alterations of ICa are probably the consequence of multifactorial effects induced by the general anesthetics. Previous suggestions have included alterations in the membrane lipid bilayer, membrane surface charge and hydrophobic pockets on channel proteins. We suggest that the increase in Cai may also participate to these effects.

Action Potentials↗
PubMed1990

Calcium current in single cells isolated from normal and hypertrophied rat heart. Effects of beta-adrenergic stimulation.

The L-type calcium current was investigated in normal and hypertrophied rat ventricular myocytes as a possible cause of the action potential lengthening that has been reported during hypertrophy. Regulation of the calcium current (ICa) by a beta-adrenergic agonist (isoproterenol) was also analyzed since beta-agonist-induced positive inotropy is less marked in hypertrophied heart. Left ventricular hypertrophy was induced by stenosis of the abdominal aorta. For recording ICa, the whole-cell patch-clamp technique was used. Potassium currents were suppressed by replacing K+ ions with Cs+ ions in both the extracellular and intracellular media, and sodium current was blocked by 50 microM tetrodotoxin. The Ca2+ current was larger in hypertrophied cells (2.2 +/- 0.6 nA [n= 31]) than in normal cells (1.2 +/- 0.5 nA [n = 33]). However, if one relates ICa amplitude to the cell membrane area, as estimated by membrane capacitance measurement, no significant difference was observed in current density (8.5 +/- 2.5 pA/pF [n = 31] and 8.3 +/- 2.1 pA/pF [n = 33] in hypertrophied and in normal cells, respectively). In both cell types, ICa displayed the same voltage and time dependence. When expressed as a percentage, the maximal increase in ICa amplitude that was obtained with 100 nM isoproterenol was less in hypertrophied cells (+78%) than in normal cells (+120%). The sensitivity of ICa to beta-adrenergic stimulation was not modified: EC50 was 3.8 nM for hypertrophied cells and 4.8 nM for normal cells. Forskolin and cyclic AMP were as effective in both cell types. Stimulation of ICa by beta-adrenergic agonist was decreased in agreement with a reduced number of binding sites of beta-agonists and/or an altered coupling of the G-proteins.

Action Potentials↗
PubMed1990

Neurohormonal control of calcium sensitivity of myofilaments in rat single heart cells.

To investigate the changes in the properties of cardiac contractile proteins due to neurohormonal stimulation, different agonists were applied to single cells isolated from rat ventricle. Cells were then rapidly skinned by Triton X-100, and force was recorded after gluing the cells to a strain gauge. The skinned cells had mechanical properties very similar to those described for thin trabeculas. Tension-pCa relations were highly reproducible from one cell to another, with sarcomere length fixed at 2.1 microns. The application of alpha 1-adrenergic and muscarinic agonists, which increase the turnover of phosphatidylinositol, for 5 minutes before skinning the cells increased the sensitivity of the myofilaments to calcium, as indicated by a leftward shift of the tension-pCa relation, whereas beta-adrenergic stimulation induced a rightward shift. The increase in calcium sensitivity was also evoked by protein kinase C activators such as 1,2-dioctanoylglycerol and phorbol 12-myristate 13-acetate but not by protein kinase C itself or by purinergic agonists, although the latter also increased the turnover of phosphatidylinositol. Incubation of the skinned cells with phosphatase reversed the alterations in calcium sensitivity induced by previous agonist stimulation of the intact cells. In conclusion, this study demonstrates a potentially influential mechanism for the physiological regulation of cardiac muscle contractility.

Actin Cytoskeleton↗
PubMed1990

The mechanism of positive inotropy induced by adenosine triphosphate in rat heart.

When applied extracellularly in the micromolar range, ATP and related compounds induced a positive inotropy in the rat papillary muscle. This was also true in the rat auricle after pertussis toxin treatment. Then, in both tissues, ATP further increased the contraction after a maximal beta-adrenergic stimulation. The increase in contractile force could be related to the increase in the calcium current. The L-type calcium current was measured by whole-cell patch-clamp recording in single cells isolated from the rat ventricle after the sodium and potassium currents were inhibited by tetrodotoxin and cesium, respectively. When added alone, 10 microM ATP increased the calcium current by 60%. Adenosine 5'-O-(3-thiotriphosphate) was also able to increase calcium current. Adenosine was much less effective, and GTP, UTP, CTP, and ITP were without effect. A similar increase in calcium current was observed when ATP was added in addition to a maximal stimulation by a beta-adrenergic agonist or after internal perfusion with cyclic AMP. However, this increase was preceded by a transient decrease whose origin could not be attributed to a P1-purinergic agonistic effect of ATP. The transient decrease was not elicited by adenosine or in a magnesium-free HEPES solution and was not suppressed after pertussis toxin treatment. This effect appeared related to the variations in the holding current also observed upon ATP application. Together with vasodilation, ATP and adenine compounds induced positive inotropy. The latter effect could be attributed in part to the increase in calcium current and was independent of cyclic AMP. Both effects are complementary with the beta-adrenergic stimulation and can help healthy cells to compensate the failing zone from which ATP could be released.

1-Methyl-3-isobutylxanthine↗
PubMed1989

Inhibition of ICa in single frog cardiac cells by quinidine, flecainide, ethmozin, and ethacizin.

The effects of four class I antiarrhythmic compounds on the Ca current (ICa), recorded with whole cell patch clamp in single isolated frog ventricular cells, were compared. Na and K currents were blocked by tetrodotoxin and Cs. Quinidine and flecainide induced an apparent tonic block with a 50% effective dose (ED50) at 10 and 20 microM, respectively; there was no clear use-dependent inhibition. Ethmozin and ethacizin, two phenothiazine derivatives, exhibited both tonic and use-dependent inhibition. Ethacizin was at least 10 times more potent than ethmozin; at 1 microM, it induced a 15% tonic block and 5, 35, and 42% use-dependent block at 0.125, 1, and 2 Hz, respectively. These compounds appeared only 3- to 10-fold less efficient on the ICa than on the Na current recorded in parallel experiments, with flecainide showing the largest different potency. All four compounds shifted the availability curves by a few millivolts toward hyperpolarization, had a clear voltage-dependent inhibition, and slowed reactivation, the latter effect being more marked with less negative holding potential. Consequently, the absence of use-dependence inhibition with quinidine and flecainide could be the consequence of a very fast association of the two compounds with the open channels that would be complete during the 200-ms depolarizing pulse. Since arrhythmias are frequently associated with tissue depolarization and can be related to Ca-dependent action potential and slow conduction, the inhibition of the ICa reported above should account, in part, both for the antiarrhythmic and the negative inotropic effects of these compounds.

Animals↗
PubMed1989

Ethacizin blockade of calcium channels: a test of the guarded receptor hypothesis.

The effect on calcium channels of the sodium channel antagonist, ethacizin, was studied in isolated frog ventricular cells using the whole cell voltage-clamp methodology. Ethacizin was found to block inward calcium current in a frequency-, voltage-, and concentration-dependent manner. The frequency-dependent blocking properties were modeled by considering the drug interaction with a voltage-dependent mixture of calcium channels harboring either an accessible or an inaccessible binding site. With repetitive stimulation, the pulse-to-pulse reduction in peak current is shown to be exponential, with a rate linearly related to the interstimulus interval and the drug concentration. Observed frequency- and concentration-dependent blocks were consistent with the predictions of the model, and mixture-specific rate constants were estimated from these data. The negligible shift in channel inactivation and the reduction of apparent binding and unbinding rates with more polarized membrane potentials imply the active moiety of ethacizin blocks open channels and is trapped within the channel at resting membrane potentials. The binding rate at 0 mV is similar to that observed in studies of interactions of other open channel blocking agents with voltage- and ligand-gated channels.

Animals↗