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J Vereecke

Publications and source records attributed to J Vereecke.

At least 37 records · Page 2Linked to original sources
PubMed1991

Effects of the enantiomers of disopyramide and its major metabolite on the electrophysiological characteristics of the guinea-pig papillary muscle.

Disopyramide, a Class Ia antiarrhythmic drug, is clinically used as a racemic mixture; R(-)disopyramide and S(+)disopyramide. The major metabolite in man is desisopropyldisopyramide: R(-)desisopropyldisopyramide and S(+)desisopropyldisopyramide. The effects of the four compounds were compared on the electrophysiological characteristics of the guinea-pig papillary muscle using the standard microelectrode technique. At an external K+ concentration of 5.4 mmol/l and a stimulation frequency of 1 Hz, S(+)disopyramide (20 mumols/l) increased action potential duration (APD) by more than 18%, while it was diminished by 6% in the presence of R(-)disopyramide. Resting membrane potential amounted to -87.1 +/- 0.5 mV (n = 14) and -85.6 +/- 1.2 mV (n = 10), respectively. Also a small but significant difference in effect on the maximal rate of depolarization was observed, R(-)disopyramide being more potent, related with a slower recovery of the maximal rate of depolarization. The enantiomers of the metabolite appeared to be three times less potent than those of the parent drug in their effect on the maximal rate of depolarization. The characteristics of the enantiomers of the metabolite correlated with those of the parent drug: also the R(-)enantiomer was more potent in decreasing the maximal rate of depolarization and caused more shortening of the action potential than the S(+)enantiomer. Time constants for onset and recovery of/from rate dependent block of the maximal rate of depolarization were dependent upon the external K+ concentration, both for the enantiomers of the parent drug and those of the metabolite. Onset slowed down while recovery accelerated when external K+ was increased. Time constants were lower for the metabolite. When stimulation interval was shortened, the effect on the maximal rate of depolarisation increased. Only for the metabolite statistical significant stereoselective differences were observed at all stimulation intervals. The effects on the action potential duration were dependent upon stimulation interval; for all enantiomers the action potential duration tended to be relatively (% of control) higher at short stimulation intervals than at large stimulation intervals. The effect on the maximal rate of depolarization was also voltage dependent, but no significant differences were observed between the enantiomers, for the parent drug as well as for the metabolite.

Action Potentials↗
PubMed1991

Stereoselective effects of the enantiomers of bupivacaine on the electrophysiological properties of the guinea-pig papillary muscle.

1 Direct myocardial effects of the S(-)- and R(+)-enantiomers of bupivacaine were compared in the guinea-pig isolated papillary muscle by recording transmembrane action potentials with the standard microelectrode technique. 2 In 5.4 mM K+, at a stimulation rate of 1 Hz, the maximal rate of depolarization (Vmax) was reduced to 59.9 +/- 1.4% (n = 10) of control (mean +/- s.e.mean) in the presence of 10 microM R(+)-bupivacaine, and to 76.7 +/- 1.2% (n = 14) in the presence of the same concentration of S(-)-bupivacaine. This was mainly due to a difference in time constant at which block dissipated during the diastolic period. Recovery was slower in the presence of R(+)-bupivacaine. The slower recovery in the presence of R(+)-bupivacaine resulted also in a more pronounced frequency-dependent block of Vmax. 3 Time constants for recovery from use-dependent block became significantly faster for both enantiomers on hyperpolarization, while no significant change was observed at depolarization. At all membrane potentials recovery was slower in the presence of R(+)-bupivacaine. 4 The action potential duration (APD) was shortened to a greater extent in the presence of R(+)-bupivacaine over a large range of stimulation frequencies. 5 We conclude that S(-)-bupivacaine affects Vmax and APD in the guinea-pig papillary muscle less than the R(+)-enantiomer at different rates of stimulation and resting membrane potentials.

Action Potentials↗
PubMed1991

Atrial membranes contain nucleoside diphosphate kinase (NDPK) activity: its role in regulation of muscarinic K+ channels.

Guinea pig or rabbit atrial muscarinic K+ channels in cell-free inside-out patches can be activated in the absence of extracellular agonist and cytoplasmic G nucleotides by intracellular ATP-Mg2+. This ATP-dependent activation is compatible with the existence of a membrane-bound nucleoside diphosphate kinase (NDPK), which directly phosphorylates GK-bound GDP. We show that this ATP-dependent activation is also possible in frog atrial cells, and that atrial membranes of frog and guinea pig contain NDPK activity. The relative order of different nucleoside triphosphates (NTPs) as phosphate donors parallels the observed efficiency of these nucleotides in activation of the channels. Thus, atrial membranes contain NDPK activity, which can be responsible for the ATP-dependent activation of muscarinic K+ channels, seen in patches of atrial cells. Under physiological conditions, NDPK can act as a GTP supply in the immediate vicinity of the G protein to ensure reliable signal transduction.

Adenosine Triphosphate↗
PubMed1991

Depolarization-induced influx of sodium in response to phenylephrine in rat atrial heart muscle.

1. The effects of alpha 1-adrenoceptor stimulation on transmembrane potential, currents and ion fluxes were investigated in multicellular preparations and/or single cells obtained from the left atrium of rat hearts. 2. In multicellular preparations, phenylephrine caused a concentration-dependent positive inotropic effect, an increase in action potential duration, and a decrease in resting potential; the effects were antagonized by phentolamine. 3. In the presence of phenylephrine (100 mumol/1), two levels of resting potential were observed when the preparations were, alternately, electrically stimulated or kept at rest (-74 +/- 1 mV during activity and -62 +/- 4 mV at rest; mean +/- S.E.M.; n = 9). 4. In resting preparations, the depolarization in response to phenylephrine was eliminated in low-Na+ solution (12 mmol/l) and antagonized by tetrodotoxin (10 mumol/l). 5. The phenylephrine-induced depolarization was also seen in nominally Ca(2+)-free solution and in the presence of (-)-devapamil (1 mumol/l). 6. The alkylating agent N-ethyl-maleimide (30 mumol/l) abolished the depolarizing effect of phenylephrine. 7. Phorbol 12,13-dibutyrate (10 mumol/l) also abolished the depolarizing effect of phenylephrine. 8. Phenylephrine caused a significant increase of 22Na+ uptake in resting preparations and of 45Ca2+ uptake in beating preparations. 9. The depolarizing effect of phenylephrine was also observed in single atrial myocytes. Steady-state membrane currents in response to 500 ms depolarizing and hyperpolarizing voltage clamp steps were decreased. The cross-over of I-V curves under control and test conditions was at about -70 mV. The effects of phenylephrine were antagonized in the presence of phentolamine. 10. After suppression of potassium currents by substitution of CsCl for internal and external KCl ([KCl]o), phenylephrine had no effect on membrane currents. 11. In conclusion, we presume the following sequence of events in response to phenylephrine in rat atrial heart muscle. First, the stimulation of alpha 1-adrenoceptors decreases the K+ conductance thereby producing a depolarization in the presence of an inward current. Second, the change of the membrane potential in the depolarizing direction induces a TTX-sensitive Na+ window current which further propels the depolarization. Third, the increase in Na+ influx may increase Ca2+ influx by activating the Na(+)-Ca2+ exchange in mechanism. The greater influx of Ca2+ may contribute to the positive inotropic effect in response to phenylephrine.

Action Potentials↗
PubMed1991

Dual effect of the local anaesthetic penticainide on the Na+ current of guinea-pig ventricular myocytes.

1. The effect of the local anaesthetic penticainide (2-alkyl-(4-dialkylamino)-2-pyridyl-butyramide) on macroscopic and single-channel sodium current (INa) of guinea-pig ventricular myocytes was studied with the patch-clamp technique in the cell-attached and inside-out mode. 2. Penticainide (3-60 microM) affected the INa from the outside as well as from the cytoplasmic side. 3. Peak INa was reduced by penticainide at concentrations of 6, 30 and 60 microM, and this decrease of peak INa was more pronounced when the holding potential was more negative. Despite a reduction of peak INa, the time integral of the Na+ current was not changed (60 microM) or was even enhanced (6 microM), and this enhancement became more pronounced at less negative potentials. 4. At a concentration of 3 microM, penticainide increased both the time integral of the current and peak INa. 5. The shape of the steady-state current-voltage relationship and the steady-state inactivation curve were not influenced by penticainide. 6. In pronase-modified inside-out patches penticainide reduced INa at the beginning of a depolarizing pulse to the same extent as at the end (400 ms), indicating a very fast blockade of the bursting Na+ channel. The most prominent effects on pronase-modified single-channel INa were an increase of sweeps without activity, and a fast, repeatedly occurring block (flickering) of the bursting Na+ channel. 7. The amplitude of the unitary current was not altered. 8. It is concluded that penticainide blocks the open Na+ channel, and in addition shows the macroscopic inactivation.

Anesthetics, Local↗
PubMed1991

Ionic currents activated during hyperpolarization of single right atrial myocytes from cat heart.

Whole-cell recording techniques were used on single right atrial myocytes to study the ionic currents that may be responsible for the diverse diastolic voltage characteristics of atrial tissue. Ionic currents were activated by hyperpolarizing voltage pulses negative to -30 mV. In general, four different types of cells were identified based primarily on the ionic currents elicited during hyperpolarization. The first cell type exhibited an inward current that decayed with time at more negative voltages, reversed near the potassium equilibrium potential, inwardly rectified at more positive voltages, increased in elevated extracellular potassium, and was blocked by 3 mM barium or 10 mM cesium. This current was identified as the potassium current iK1. A second cell type exhibited a time-dependent inward current that increased at more negative voltages, had an activation range between -50 and -110 mV, had a reversal potential of -26 mV, and was blocked by 3 mM cesium. This current was identified as an if current. A third cell type exhibited an inward current that initially decayed and then became more inward with time. Barium (3 mM) abolished the initial inward current and revealed a time-dependent increasing inward current that was blocked by 3 mM cesium. This current was composed of both the iK1 and if currents. A fourth cell type exhibited only small time-independent leak currents in response to hyperpolarization. These results indicate that individual cells within the right atrium are electrophysiologically heterogeneous with respect to the types of ionic channels present in their sarcolemmal membranes. This specialization in ionic currents partially explains the diverse diastolic voltage characteristics and functional properties of atrial tissue.

Action Potentials↗
PubMed1990

Non-selective cation and dysfunctional chloride channels in the apical membrane of nasal epithelial cells cultured from cystic fibrosis patients.

Chloride channels and non-selective cation channels in the apical membranes of cultured nasal epithelial cells from three cystic fibrosis patients were investigated with the patch-clamp technique. Outwardly rectifying chloride channels were found in 31% of the inside-out patches, but activity of this channel was never observed in cell-attached patches, even after stimulation with adrenaline. In 30% of the patches with chloride channels, activation occurred immediately after excision. Most of the channels, however, activated only after a membrane depolarization of +40 to +120 mV. Once activated, the chloride channels were indistinguishable from those in nasal epithelial cells of control patients. Amiloride-insensitive, calcium- and voltage-dependent, non-selective cation channels were present in 11% of the cell-attached and 43% of the cell-free patches and could not be distinguished from those in controls. The cystic fibrosis chloride channel defect is conserved in cultured nasal epithelial cells, while a non-selective cation channel is apparently not affected.

Adolescent↗
PubMed1990

ATP-dependent activation of atrial muscarinic K+ channels in the absence of agonist and G-nucleotides.

While opening of cardiac muscarinic K+ channels is mediated by a guanine nucleotide-binding protein, GK, and normally requires both ACh and cytosolic GTP, we demonstrate that in the absence of agonist and G-nucleotides guinea-pig atrial muscarinic K+ channels are activated by cytosolic ATP (K50 = 224 microM). This activation involves a phosphorylating reaction and is most probably related to a transphosphorylation by a nucleoside diphosphate kinase (NDPK) from ATP to GK-bound GDP. This ATP-induced response is completely inhibited by nanomolar concentrations of GDP or GTP, suggesting a G-nucleotide specific binding site on NDPK.

Acetylcholine↗
PubMed1990

A combined study of sodium current and T-type calcium current in isolated cardiac cells.

Sodium currents (INa) and T-type calcium currents (ICa,T) of isolated guinea-pig ventricular myocytes were recorded using the whole-cell voltage-clamp technique. Separation of the two currents was obtained by using the difference current method in the presence and absence of 2 mM extracellular Na (Nao). Time to peak and the time constant of inactivation of. INa were about 5 times faster than that of ICa,T (test potential -30 mV), and ICa,T had an activation range positive to -50 mV, were inactivated at -50 mV, and their current/voltage relationships peaked at -22.3 +/- 1.8 mV (n = 18) and -29.3 +/- 0.5 mV (n = 18) respectively, with a reversal potential of +40.3 +/- 4 mV (n = 18) and +30 +/- 10 mV (n = 18), respectively [2 mM Nao; 5.4 mM extracellular Ca (Cao)]. INa was blocked by 30 microM tetrodotoxin (TTX), 500 microM lidocaine, partly inhibited by 1 mM amiloride, but not affected by 100 microM nickel (Ni). ICa,T was neither affected by 30 microM TTX nor 500 microM lidocaine, but blocked by 100 microM Ni, 1 mM amiloride, 10 microM R 56865 and use-dependently reduced by 5 microM flunarizine. Adenosine (500 microM) affected neither INa nor ICa,T, whereas 1 microM isoprenaline did not affect ICa,T, but slightly increased INa. Our results demonstrate that the characteristics of ICa,T are not affected by the concomitant activation of INa, and vice versa. We conclude that ICa,T are not Ca currents through Na channels.

Adenosine↗
PubMed1990

Outward-rectifying chloride channels in cultured adult and fetal human nasal epithelial cells.

The patch-clamp technique was used to characterize ion channels in the apical membranes of cultured human nasal epithelial cells, dissociated from fetal nasal mucosa and from adult nasal polyps. Outward-rectifying chloride channels were found in 4.3% of the cell-attached patches from fetal cells (n = 258) and in 3.1% of the patches from adult cells (n = 320). After excision the number of patches containing active chloride channels increased threefold to 13% of the patches from the fetal cells and 10% from adult cells. The single-channel conductance at 0 mV in symmetrical 150 mM NaCl solutions was 24.3 +/- 0.9 pS (n = 28) and 26.0 +/- 1.2 pS (n = 30), respectively, in adult and fetal cells and showed outward rectification in the potential range from -80 to +80 mV. In fetal cells as well as in adult cells the channels were anion selective, and were almost impermeable for larger anions and monovalent cations. In cell-free patches the channels were Ca2+ independent. In most of the channels the open probability was voltage independent and high (+/- 0.86); in 20% of the channels, however, the open probability increased with depolarization. In conclusion, fetal nasal epithelial cells contain chloride channels in their apical membranes with single-channel properties and regulatory mechanisms similar to those found in cells from adults.

Cells, Cultured↗
PubMed1990

Identification of a voltage- and calcium-dependent non-selective cation channel in cultured adult and fetal human nasal epithelial cells.

The apical membranes of cultured human nasal epithelial cells from adults and fetuses were investigated with the patch-clamp technique. Amiloride-insensitive, calcium- and voltage-dependent, non-selective cation channels were found in 4% of the cell-attached, and 18% of the inside-out and outside-out patches (n = 412). Multiple functional channels were present in more than 90% of these patches, with a mean of 3.9 channels per patch (n = 55). The current-voltage relationship can be described by the Goldman equations and the single channel conductance was 20.1 +/- 0.3 pS (n = 29) in adult and 20.7 +/- 0.4 pS (n = 44) in fetal cells in symmetrical 150 mM NaCl solutions. The channels were highly selective for cations: P Na/P Cl was 30 in adult and 45 in fetal experiments. They were equally permeable for K+ and Na+, somewhat less for Cs+, and impermeable for choline+ and tetraethylammonium+. The open probability was voltage dependent: it increased approximately 2-fold with 30 mV depolarization in the potential range from -60 mV to +60 mV. The channels were activated by Ca2+ concentrations of about 10(-4) M at the cytoplasmic side, but were insensitive to extracellular Ca2+ and amiloride (10(-4) M). The non-selective cation channels found in apical membranes of cultured fetal nasal epithelial cells were not different from the adult ones.

Adult↗
PubMed1990

Three different potassium channels in human atrium. Contribution to the basal potassium conductance.

We applied the cell-attached and inside-out patch-clamp technique under symmetrical isotonic potassium conditions on single human (and guinea pig) atrial cells. The human cells were isolated by a modified method to that described earlier. Our aim was twofold: 1) to study the single-channel characteristics of potassium channels in human atrial single cells, present under basal conditions (iK1 and iK(ATP] or when stimulated with 10(-5) M acetylcholine; and 2) to calculate the contribution of these three channel types to the total basal potassium conductance in human atrial cells, and to compare the results with data on guinea pig atrial cells under the same conditions. We found that in human cells 58% of the patches (n = 42/74) contained acetylcholine-sensitive potassium channels: their conductance was 42 +/- 1.2 pS and mean open time (tau o) was 1.7 +/- 0.5 msec. They showed sporadic openings in the absence of agonist, and activation by acetylcholine was G-protein dependent. In 16% of the patches (n = 7/44), adenosine (10(-4) M) activated the same channels, but the activity was lower than when stimulated by acetylcholine. In 18% of the patches (n = 9/51), an iK1 channel was present (conductance, 27 pS; tau o, 8.7 msec), whereas in the cell-attached mode, ATP-dependent channels were never seen. However, they were present in half of the inside-out patches on washout of ATPi (conductance, 73 pS; tau o, 1.4 msec). The basal potassium conductance (i.e., in the absence of any exogenous hormone or neurotransmitter) was mainly due to iK1 channels in both human and guinea pig cells, a finding that is in contrast with previous reports. However, the potassium current that is induced by acetylcholine is much higher in guinea pig than in human isolated cells; a fraction of it would suffice to fully determine the resting potassium conductance in guinea pig atrial cells, whereas it can play only a modulatory role in human cells. This difference could be important in species-specific autonomic modulation and antiarrhythmic drug action.

Acetylcholine↗
PubMed1990

Mechanism of cardiac T-type Ca channel blockade by amiloride.

Whole cell clamp experiments were used to study the mechanism of action of amiloride on cardiac T-type Ca channels in guinea pig ventricular myocytes. Two hundred fifty microM amiloride blocked the T-type Ca channel for approximately 50%, whereas this concentration did not suppress the L-type Ca channel. The Kd value for the block of the T-type Ca channel was 233 microM and the Hill coefficient was 1.1. The drug showed a rapid onset of action and a quick wash out with complete reversibility. Amiloride blocked the T-type Ca channel without frequency-dependency, i.e., block was established at the holding potential (-90 mV) without being dependent on activation and inactivation of the channel. Amiloride reduced the amplitude of the T-type Ca current at all potentials without changing steady-state activation and inactivation. Amiloride did not affect the T-type Ca current when the drug was applied intracellularly via the pipette solution. In summary, amiloride blocked the T-type Ca channel in a channel state- and voltage-independent way.

Amiloride↗
PubMed1989

Single-channel analysis of a large conductance channel in peroxisomes from rat liver.

Native membranes and Triton X-100 solubilized integral membrane proteins of peroxisomes from rat liver were reconstituted in liposomes. With the patch clamp technique, a channel was detected with a conductance of 420 +/- 30 pS and a PK/PC1 of about 3. The channel in native membrane fractions was weakly voltage dependent, residing most of the time in an open state with the possibility to shift to different substates. Solubilization changed the kinetic properties. The channel became strongly voltage dependent and closed at voltages negative to -20 mV. The estimated diameter of the channel is about 1.7 nm and might explain, at least partially, the permeability properties of the peroxisomal membrane.

Animals↗
PubMed1989

Properties of the bursting Na channel in the presence of DPI 201-106 in guinea-pig ventricular myocytes.

Single Na channel currents were measured in cell-attached patches of guinea-pig ventricular myocytes in the presence of the S-enantiomer of DPI 201-106. DPI changes the kinetic pattern of channel activity from short living openings at the beginning of a depolarizing pulse (voltage-independent mean open time about 0.4 ms between -60 and -20 mV), into longlasting burst of openings. The single channel current-voltage relation can be approximated by a straight line with a single channel conductance of 15 pS, which is the same as in the absence of DPI, and a reversal potential near the estimated Na equilibrium potential (+74 mV). The ensemble averaged Na current shows a fast peak of inward current, which partially decays within less than 10 ms, but which shows a large component which decays very slowly with a time constant of the order of 1 s (1.31 +/- 0.6 s at -30 mV, 19 measurements in 12 cell-attached patches). The slowly decaying component activates with a half-maximum potential at -55.4 +/- 2.3 mV and a slope parameter s of 4.9 +/- 1.9 mV. The half-maximum potential of the steady-state inactivation is -115.6 +/- 1.8 mV, and the slope parameter is 9.1 +/- 1.5 mV. The open time distribution can be fitted by a single exponential only at potentials negative to -40 mV. The time constant is 1.3 +/- 0.14 ms at -50 mV (7 patches).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Different conductance states of the bursting Na channel in guinea-pig ventricular myocytes.

Single channel currents flowing through Na channels were studied in cell-attached patches from guinea-pig ventricular myocytes after removal of the fast component of inactivation by the S enantiomer of DPI 201-106. In addition to openings with a single channel conductance of 15 pS, two other types of events occur. A low conductance state has a single channel conductance of 5 pS, while a medium conductance state can sometimes be seen at 8 pS. These different conductance states can coexist in the same patch. The low conductance state appears to have the same kinetic properties as the high conductance state. However it is less sensitive to block by TTX. Averaged currents obtained from exclusively low conductance openings also show a decay (time constant at -30 mV of 0.8 +/- 0.4 s) which is not significantly different from the decay of the high conductance channel. The average open and closed times are also similar to the values for the high conductance channel. The medium conductance state has a single channel conductance of 8 pS and a reversal potential which is similar to the high conductance channel. Closings of the high conductance state to the medium conductance state can be observed as well as openings from the medium to the high conductance state. The average current of sweeps showing only the medium conductance state has the same time course and shows a similar TTX-sensitivity as the average current of the high conductance state. Also the voltage range for steady-state activation and inactivation for the medium conductance channel matches that of the high conductance channel.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Properties of the block of single Na+ channels in guinea-pig ventricular myocytes by the local anaesthetic penticainide.

1. The blocking mechanism of a disopyramide derivative Penticainide (2-alkyl-(4-(dialkylamino)-2-)pyridyl-butyramide) on cardiac Na+ channels has been studied using single-channel analysis in cell-attached and inside-out patches from guinea-pig ventricular cells. Penticainide was applied in concentrations between 3 and 100 microM. The S-enantiomer of DPI 201-106 (5 microM) was used as a tool to slow the inactivation, improve the time resolution by prolonging the mean open time, and to increase the number of openings per depolarization of the channel. 2. When in cell-attached or inside-out patch experiments up to 100 microM-Penticainide was applied to the bathing solution no significant effect was observed on the probability of the channel being open or on the mean open time. 3. In cell-attached patch experiments with 100 microM-Penticainide in the pipette, the open-state probability of the Na+ channel was much lower than in the absence of Penticainide. No significant changes were found in the potential of half-maximum activation or in the slope of the activation curve. The maximum open-state probability was reduced by a factor five in the presence of Penticainide. The single-channel conductance was not affected by the drug. 4. The decrease in the probability of the channel being open was mainly due to an increased probability of observing sweeps with no activity ('nulls'). 5. A dramatic relief from the block was observed when pauses were interposed into the normal activation pattern, or when the pacing rate was reduced. 6. The distribution of the open times of the bursting Na+ channel could be fitted with two exponentials. Penticainide in the patch pipette reduced the mean open time. Also the contribution of the number of long openings to the total number of openings was reduced. 7. Closed-time distribution was also fitted with two exponentials. Penticainide in the patch pipette prolonged the long mean closed time. The contribution of the number of short closings to the total number of closings was decreased. 8. Penticainide in the patch pipette did not significantly change the time constant of the decay of the ensemble-averaged currents measured at -30 mV. Because of the close correlation between the burst duration and the time constant of inactivation (Nilius, Vereecke & Carmeliet, 1988b), we conclude that no striking effect of Penticainide on the burst duration can be expected.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗