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Y Kurachi

Publications and source records attributed to Y Kurachi.

At least 145 records · Page 8Linked to original sources
PubMed1990

Voltage-dependent inhibition of the delayed outward potassium current by OPC-8490, a novel positive inotropic agent, in isolated atrial myocytes of guinea-pig heart.

To elucidate the ionic mechanisms underlying prolongation of the action potential by OPC-8490, a novel positive inotropic agent, we examined effects of the drug on the membrane currents in isolated atrial myocytes of guinea-pig heart. The tight-seal whole cell clamp technique was used. In the current clamp condition, OPC-8490 prolonged the atrial action potential in a concentration-dependent fashion. In the voltage clamp condition, OPC-8490 affected neither the conventional L type Ca current nor the outward IK1 current significantly. The drug blocked the delayed outward K current (IK) in a concentration-dependent fashion. At a constant concentration of the drug, inhibition of IK became more marked as the membrane potential became more negative. The tail current of IK could be fitted by a sum of two exponentials (fast and slow components). OPC-8490 blocked the fast component more prominently than the slow one. Time constants of both components were not affected significantly by the drug. In conclusion, OPC-8490 prolongs the cardiac action potential mainly by blocking IK in a voltage-dependent and time-dependent fashion.

Action Potentials↗
PubMed1990

Properties of membrane currents in isolated smooth muscle cells from guinea-pig trachea.

Tracheal smooth muscle cells were enzymatically isolated from guinea-pig trachea. These cells contracted in response to acetylcholine (0.01-10 microM) in a concentration-dependent fashion. Under current-clamp conditions with 140 mM K+ in the pipette solution, the membrane potential oscillated spontaneously at around -30 mV. Under voltage-clamp conditions, there appeared spontaneous but steady oscillations of outward current (IO). On depolarization from a holding potential at -40 mV, three components of outward current were elicited: transient outward current (IT), steady-state outward current (IS) and IO. These three components of outward current reversed around the K+ equilibrium potential and were abolished by Cs+ in the pipette, indicating that K+ was the major charge carrier of these outward currents. All these three components were completely suppressed by extracellular tetraethylammonium (10 mM). Both IT and IO were depressed by quinidine (1 mM), 4-aminopyridine (10 mM) and nifedipine (100 nM), but IS was not affected. IT and IO were suppressed by a Ca2(+)-free perfusate with less than 1 nM Ca2+ in the pipette, while with 10 nM Ca2+ in the pipette, only IO was suppressed. In both conditions, IS was not affected by the Ca2(+)-free perfusate. Therefore, it is suggested that IO, IT and IS are separate types of K+ current. With Cs+ in the pipette, K+ currents were almost completely suppressed and a transient inward current was observed during depolarizing pulses. The inward current was not affected by tetrodotoxin and increased when the concentration of extracellular Ca2+ was raised, indicating that the current is a Ca2+ channel current. Even with a holding potential of -80 mV, the low-threshold inward current could not be observed. The high-threshold Ca2+ current was abolished by nifedipine (100 nM) and was enhanced by Bay K 8644 (100 nM). The order of permeation of divalent cations through the Ca2+ channel was Ba2+ greater than Sr2+ approximately Ca2+. Cd2+ blocked the Ca2+ current more effectively than Ni2+. These results may indicate that the Ca2+ current of tracheal smooth muscle cells is mainly composed of the current through an L-type Ca2+ channel.

4-Aminopyridine↗
PubMed1990

Positive cooperativity in activation of the cardiac muscarinic K+ channel by intracellular GTP.

Concentration-dependent effects of intracellular GTP on activation of the muscarinic K+ channel were examined in inside-out patches of cardiac atrial myocytes. The pipette solution contained 0.1 microM ACh. GTP (0.01-30 microM) and 0.5 mM MgCl2 were applied to the inside side of the patch membrane. K+ channels were activated with GTP concentration above 0.1 microM. Channel activation reached a maximal value with 1-3 microM GTP. It decreased at GTP concentrations larger than 3 microM, probably due to desensitization. The dependence of the open probability of the channel on intracellular GTP showed a sigmoidal relationship with a Hill coefficient of around 3. A positive cooperative effect of intracellular GTP on the K+ channel may play an important role in amplifying the signal from the membrane receptor to the K+ channel.

Animals↗
PubMed1990

Adenosine-5'-triphosphate-induced sinus tachycardia mediated by prostaglandin synthesis via phospholipase C in the rabbit heart.

Effects of adenosine 5'-triphosphate (ATP) and adenosine on cardiac sinus pacemaker activity were examined in the rabbit heart. Electrocardiograms of hearts were recorded while using the Langendorff perfusion method. Both adenosine and ATP, added to the perfusate, slowed the sinus pacemaker activity in a concentration-dependent manner. But in about 40% of the cases. ATP higher than 300 microM initially accelerated and then slowed the heart. The sinus slowing caused by adenosine and ATP was blocked by theophylline (a P1 receptor antagonist) and disappeared in the hearts pre-treated with islet-activating protein (1AP). In contrast, the ATP-induced sinus acceleration was not affected by either theophylline or IAP. In about 75% of the IAP-treated hearts. ATP persistently accelerated the sinus pacemaker. In the remaining 25% of the hearts, ATP caused junctional tachycardia, which may have masked the ATP-induced sinus acceleration. Apamin specifically blocked the ATP-induced sinus acceleration, suggesting that P2 receptors are involved. Among various adenine nucleotide analogues, the order of potency in inducing tachycardia in IAP-treated hearts is adenosine-5'-[gamma-thio]triphosphate greater than adenylyl imidodiphosphate greater than adenosine 5'-[alpha, beta-methylene]triphosphate = ATP greater than adenosine diphosphate = adenosine 5'-[beta, gamma-methylene]triphosphate. ATP-induced acceleration was partially blocked by indomethacin and aspirin (cyclooxygenase inhibitors), but not by nordihydroguaiaretic acid (a lipoxygenase inhibitor). These results suggest that cyclooxygenase and not lipoxygenase metabolites of arachidonic acid, e.g. prostaglandins, may be involved in the generation of tachycardia.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗
PubMed1990

Heparin uncouples the muscarinic receptors from GK protein in the atrial cell membrane of the guinea-pig heart.

The effects of heparin on activation of the G protein-gated muscarinic K+ channel were examined in atrial cells of guinea-pig heart. The inside-out patch clamp technique was used. The pipette solution contained 1.1 microM acetylcholine (ACh). In the inside-out patches, intracellular GTP activated the muscarinic, K+ channel. When heparin (0.05-5 units/ml) was further added to the intracellular side of the patch membrane, the channel openings were depressed in a concentration-dependent fashion. The effects of heparin were reversible after wash-out. Heparin did not affect GTP-gamma S-induced activation of the K+ channel. Therefore, it is suggested that heparin may uncouple the muscarinic receptors from GK protein in the cardiac atrial cell membrane.

Animals↗
PubMed1990

Trimebutine maleate has inhibitory effects on the voltage-dependent Ca2+ inward current and other membrane currents in intestinal smooth muscle cells.

We examined effects of trimebutine maleate on the membrane currents of the intestinal smooth muscle cells by using the tight-seal whole cell clamp technique. Trimebutine suppressed the Ba2+ inward current through voltage-dependent Ca2+ channels in a dose-dependent manner. The inhibitory effect of trimebutine on the Ba2+ inward current was not use-dependent. It shifted the steady-state inactivation curve to the left along the voltage axis. Trimebutine also had inhibitory effects on the other membrane currents of the cells, such as the voltage-dependent K+ current, the Ca2(+)-activated oscillating K+ current and the acetylcholine-induced inward current. These relatively non-specific inhibitory effects of trimebutine on the membrane currents may explain, at least in part, the dual actions of the drug on the intestinal smooth muscle contractility, i.e. inhibitory as well as excitatory.

Animals↗
PubMed1990

Glibenclamide specifically blocks ATP-sensitive K+ channel current in atrial myocytes of guinea pig heart.

Effects of glibenclamide on the control membrane ionic currents, acetylcholine or adenosine-induced K+ current, and nicorandil-induced K+ current were examined in single atrial myocytes of guinea pig heart. The nystatin-whole cell clamp technique was used. Nicorandil evoked the time-independent K+ current which is probably the current through the ATP-sensitive K+ channel. Glibenclamide inhibited this current in a concentration-dependent fashion, although it had no effect on the other currents. We concluded that glibenclamide specifically inhibits the ATP-sensitive K+ channel current in cardiac myocytes.

Adenosine Triphosphate↗
PubMed1990

Biosynthesis and functions of leukotriene C4.

1. The LTC4 synthase activity is rich in the microsomal fraction of the guinea pig spleen and lung. The enzyme was partially purified from the guinea pig lung and separated from the microsomal glutathione S-transferase (GST), by column chromatograpy. The enzyme has a specific activity of 40 nmol/min.mg, and acts preferentially on 5, 6-LTA4. Various types of cytosolic GSTs utilize all types of LTA4 isomers (5,6-, 11,12- and 14,15-LTA4) almost to the same extent, and methyl ester forms are better substrates for GST. 2. Two different types of GSTs (Yn1n1 and P) were purified from rat brain cytosol, to homogeneity. Because both types have a high LTC4 synthase activity, they may participate in the LTC4 production in the rat brain. 3. LTC4, produced in the guinea pig atrium, stimulates pertussis toxin (IAP)-sensitive muscarinic K+ channel (IK.ACh). The negative chronotropic action of alpha 1-adrenergic agonist might relate to the production of arachidonate lipoxygenase metabolites. These results together with the findings in Aplysia sensory neurons, suggest a novel mode of eicosanoid actions.

Animals↗
PubMed1989

AN-132, a new class I anti-arrhythmic agent, depresses the acetylcholine-induced K+ current in atrial myocytes.

AN-132, a newly developed anti-arrhythmic agent, effectively depressed the acetylcholine (ACh)-induced K+ current (IK.ACh), as measured with the tight-seal, whole-cell clamp technique in single atrial cells of guinea-pig. When GTP-gamma S was loaded into the cell through a pipette, IK.ACh was activated persistently, probably due to irreversible activation of G proteins by GTP-gamma S. AN-132 was much less potent to depress IK.ACh in GTP-gamma S-loaded cells than in GTP-loaded cells. In conclusion, AN-132 may block mainly the cardiac muscarinic ACh receptors to suppress IK.ACh.

Acetylcholine↗
PubMed1989

Arachidonic acid metabolites as intracellular modulators of the G protein-gated cardiac K+ channel.

Arachidonic acid is released from cell membranes in response to receptor-dependent as well as receptor-independent stimulation in various cells, including cardiac myocytes. Arachidonic acid is converted to prostaglandins by cyclooxygenase and to leukotrienes by 5-lipoxygenase, metabolites which are very biologically active and modulate cellular functions such as platelet aggregation, smooth muscle contraction and neural excitation. The molecular mechanisms underlying their modulations are, however, still badly understood. Here, we report that the 5-lipoxygenase metabolites of arachidonic acid activate the pertussis toxin-sensitive G protein-gated muscarinic K+ channel (IK.ACh): arachidonic acid activation of IK.ACh was prevented by the lipoxygenase inhibitors, nordihydroguaiaretic acid and AA-861; leukotriene A4 and C4 activated IK.ACh. The activation occurred in pertussis toxin-treated atrial cells and ceased when inside-out patches were formed but the patches were still susceptible to stimulation by GTP and to inhibition by GDP-beta-S. These results indicate that arachidonic acid metabolites may stimulate the G-protein in a receptor-independent way.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗
PubMed1989

Cardiac Ca current does not run down and is very sensitive to isoprenaline in the nystatin-method of whole cell recording.

The conventional L type Ca2+ channel current (ICa.L) was measured in single atrial and ventricular myocytes, with a new whole-cell recording technique using an ionophore, nystatin. The membrane of a cell-attached patch was gradually permeabilized by nystatin (100-200 micrograms/ml), added to the pipette solution, within 2-5 min after formation of a G omega-seal. The electrical activity of the cells was measured through the pipette. ICa.L, measured with the nystatin-whole cell recording technique, did not exhibit the so-called "run-down" phenomenon for up to 90 min. The response of ICa.L to isoprenaline was also well preserved during the measurement. The half maximal concentration for the isoprenaline-induced increase of ICa.L was 8.2 x 10(-9) M, which is a much smaller value than that reported previously. Thus, the nystatin-whole cell clamp recording is a useful technique to measure membrane currents of cardiac myocytes with preserving the physiological intracellular milieu.

Animals↗
PubMed1989

Activation of atrial muscarinic K+ channels by low concentrations of beta gamma subunits of rat brain G protein.

Effects of G protein beta gamma subunits from rat brain on cardiac K+ channel was examined in single atrial cells of guinea-pig, using patch clamp techniques. We found that 10 pM concentration of rat brain beta gamma subunits preparation could activate the atrial muscarine receptor-gated K+ channel (IK.ACh). Neither the detergent, CHAPS, used to suspend beta gamma nor the boiled beta gamma preparation activated IK.ACh. Furthermore, preincubation of beta gamma subunits preparation in Mg2+-free solution, which easily inactivated alpha-GTP-gamma S, did not affect beta gamma-activation of IK.ACh. We concluded, therefore, that beta gamma subunits themselves can activate IK.ACh.

Acetylcholine↗
PubMed1989

Alpha-adrenergic activation of the muscarinic K+ channel is mediated by arachidonic acid metabolites.

Phenylephrine (10 microM), added in the bathing solution, stimulated the cardiac muscarinic K+ channel (IK.ACh) in the cell-attached patch. The pipette solution contained 10 microM atropine and 100 microM theophylline to block the muscarinic acetylcholine and adenosine receptors, respectively. The channel activation induced by phenylephrine was blocked by prazosin, an alpha 1-antagonist, indicating that alpha 1-adrenergic receptor mediates the response. Phenylephrine-induced activation was prevented by nordihydroguaiaretic acid, a lipoxygenase inhibitor, and AA-861, a 5-lipoxygenase inhibitor, but was not affected by indomethacin, a cyclooxygenase inhibitor. These observations suggest that 5-lipoxygenase metabolites of arachidonic acid may be involved in the alpha-adrenergic activation of IK.ACh.

Animals↗
PubMed1989

Effect of adenosine and adenosine analogs on [14C]aminopyrine accumulation by rabbit parietal cells.

Adenosine receptors that modulate adenylate cyclase activity have been identified recently in a number of tissues. Adenosine A2 receptor is stimulatory to adenylate cyclase, whereas adenosine A1 receptor is inhibitory to adenylate cyclase. We investigated the effect of adenosine and its analogs on [14C]aminopyrine accumulation by rabbit parietal cells. Rabbit gastric mucosal cells were isolated by enzyme digestion. Parietal cells were enriched by nonlinear percoll gradients. [14C]Aminopyrine accumulation was used as an indicator of acid secretion. The effect of 2-chloroadenosine on histamine-stimulated [14C]aminopyrine accumulation was studied. The effects of N-ethylcarboxamideadenosine, 2-chloroadenosine, stable analogs of adenosine, and adenosine on [14C]aminopyrine accumulation were assessed. Cyclic AMP content of parietal cells was determined by radioimmunoassay. Histamine and carbachol, known secretagogues, stimulated [14C]aminopyrine accumulation. 2-Chloroadenosine did not suppress histamine-stimulated [14C]aminopyrine accumulation. 2-Chloroadenosine, N-ethylcarboxamideadenosine, and adenosine dose dependently increased [14C]aminopyrine accumulation. The order of potency was N-ethylcarboxamideadenosine greater than 2-chloroadenosine greater than adenosine. 8-Phenyltheophylline and theophylline, adenosine-receptor antagonists, or cimetidine did not have significant effects on the increase of AP uptake induced by 2-chloroadenosine. Coadministration of dipyridamole, and adenosine uptake inhibitor, augmented the effect of adenosine on [14C]aminopyrine accumulation. 2-Chloroadenosine, N-ethylcarboxamideadenosine, and adenosine each induced a significant increase in cellular cyclic AMP. We conclude that there may be adenosine A2 receptors on rabbit parietal cells which modulate gastric acid secretion.

2-Chloroadenosine↗
PubMed1989

Anti-cholinergic effects of quinidine, disopyramide, and procainamide in isolated atrial myocytes: mediation by different molecular mechanisms.

Effects of quinidine, disopyramide, and procainamide on the acetylcholine (ACh)-induced K+ channel current were examined in single atrial cells, using the tight-seal, whole-cell clamp technique. The pipette solution contained guanosine-5'-triphosphate (GTP) or guanosine-5'-O-(3-thiotriphosphate) (GTP-gamma S, a nonhydrolysable GTP analogue). In GTP-loaded cells, not only ACh but also adenosine induced a specific K+ channel current via GTP-binding proteins (G) by activating muscarinic ACh or adenosine receptors. Quinidine and disopyramide depressed the ACh-induced K+ current quite effectively. Procainamide had a weak inhibitory effect. Quinidine also depressed adenosine-induced K+ current, while the effect of disopyramide on adenosine-induced current was much smaller than that on ACh-induced current. In GTP-gamma S-loaded cells, the K+ channel was uncoupled from the receptors and was activated irreversibly, probably due to direct activation of G proteins by GTP-gamma S. Quinidine depressed the GTP-gamma S-induced K+ current just as in the cases of ACh- and adenosine-induced currents of GTP-loaded cells. Disopyramide had only a weak inhibitory effect and procainamide showed no effect. From these results, it is strongly suggested that the major mechanisms underlying the anti-cholinergic effects of quinidine, disopyramide, and procainamide are different; quinidine may inhibit the muscarinic K+ channel itself and/or G proteins, while disopyramide and high doses of procainamide may mainly block functions of muscarinic ACh receptors in atrial myocytes.

Animals↗