• Korean Journal of Veterinary Research
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• v.41 no.4
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• pp.485-495
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• 2001

It was investigated whether $Ca^{2+}$ and $K^+$ channels were involved in the inhibitory action of nitric oxide (NO) on the contractile and slow wave activity of guinea pig gastric antral circular muscle. The gastric antral circular muscle showed spontaneous phasic contraction and slow wave. NO donors, 3-morpholinosydnonimine hydrochloride (SIN-1, $0.01{\sim}100{\mu}M$) and S-nitroso-L-cysteine (CysNO, $0.001{\sim}10{\mu}M$), reduced not only the amplitude of phasic contraction but also that of slow wave in a concentration-dependent manner. Both the perfusion of $Ca^{2+}$-free solution and the administration of $Ni^{2+}$, a nonselective $Ca^{2+}$ channel blocker, reduced the phasic contraction as well as the amplitude and frequency of the slow wave. The effects of these treatments were similar to those of NO donors. Nifedipine ($10{\mu}M$), a specific L-type $Ca^{2+}$ channel blocker, abolished the phasic contraction and remarkably reduced the plateau of slow wave but had no profound effect on the upstroke of slow wave. In the whole-cell patch clamp mode, CysNO shifted the steady-state activation curve for L-type $Ca^{2+}$ current to the right and the steady-state inactivation curve to the left. Pretreatment of various $K^+$ channel blockers such as tetraethylammonium (1 mM), 4-aminopyridine (0.5 mM), glibenclamide (10 mM), apamin ($0.1{\mu}M$), and iberiotoxin ($0.1{\mu}M$) did not affect the inhibitory action of SIN-1. These results suggest that NO donors suppress mechanical and electrical activity of guinea pig gastric antral circular muscle by inhibition of L-type $Ca^{2+}$ channel rather than by activation of $K^+$ channels.

• The Korean Journal of Physiology and Pharmacology
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• v.14 no.1
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• pp.29-35
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• 2010

We have shown that myosin light chain kinase (MLCK) was required for the off-contraction in response to the electrical field stimulation (EFS) of feline esophageal smooth muscle. In this study, we investigated whether protein kinase C (PKC) may require the on-contraction in response to EFS using feline esophageal smooth muscle. The contractions were recorded using an isometric force transducer. On-contraction occurred in the presence of $N^G$-nitro-L-arginine methyl ester (L-NAME), suggesting that nitric oxide acts as an inhibitory mediator in smooth muscle. The excitatory composition of both contractions was cholinergic dependent which was blocked by tetrodotoxin or atropine. The on-contraction was abolished in $Ca^{2+}$-free buffer but reappeared in normal $Ca^{2+}$-containing buffer indicating that the contraction was $Ca^{2+}$ dependent. 4-aminopyridine (4-AP), voltage-dependent $K^+$ channel blocker, significantly enhanced on-contraction. Aluminum fluoride (a G-protein activator) increased on-contraction. Pertussis toxin (a $G_i$ inactivator) and C3 exoenzyme (a rhoA inactivator) significantly decreased on-contraction suggesting that Gi or rhoA protein may be related with $Ca^{2+}$ and $K^+$ channel. ML-9, a MLCK inhibitor, significantly inhibited on-contraction, and chelerythrine (PKC inhibitor) affected on the contraction. These results suggest that endogenous cholinergic contractions activated directly by low-frequency EFS may be mediated by $Ca^{2+}$, and G proteins, such as Gi and rhoA, which resulted in the activation of MLCK, and PKC to produce the contraction in feline distal esophageal smooth muscle.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.6
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• pp.471-477
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• 2000

In the rabbit renal artery, acetylcholine $(ACh,\;1\;nM{\sim}10\;{\mu}M)$ induced endothelium-dependent relaxation of arterial rings precontracted with norepinephrine $(NE,\;1\;{\mu}M)$ in a dose-dependent manner. $N^G-nitro- L-arginine$ (L-NAME, 0.1 mM), an inhibitor of NO synthase, or ODQ $(1\;{\mu}M),$ a soluble guanylate cyclase inhibitor, partially inhibited the ACh-induced endothelium-dependent relaxation. The ACh-induced relaxation was abolished in the presence of 25 mM KCl and L-NAME. The cytochrome P450 inhibitors, 7- ethoxyresorufin $(7-ER,\;10\;{\mu}M),$ miconazole $(10\;{\mu}M),$ or 17-octadecynoic acid $(17-ODYA,\;10\;{\mu}M),$ failed to inhibit the ACh-induced relaxation in the presence of L-NAME. 11,12-epoxyeicosatrienoic acid $(11,12-EET,\;10\;{\mu}M)$ had no relaxant effect. The ACh-induced relaxation observed in the presence of L-NAME was significantly reduced by a combination of iberiotoxin $(0.3\;{\mu}M)$ and apamin $(1\;{\mu}M),$ and almost completely blocked by 4-aminopyridine (5 mM). The ACh-induced relaxation was antagonized by $P_{2Y}$ receptor antagonist, cibacron blue $(10\;and\;100\;{\mu}M),$ in a dose-dependent manner. Furthermore, 2-methylthio-ATP (2MeSATP), a potent $P_{2Y}$ agonist, induced the endothelium-dependent relaxation, and this relaxation was markedly reduced by either the combination of iberiotoxin and apamin or by cibacron blue. In conclusion, in renal arteries isolated from rabbit, ACh produced non-NO relaxation that is mediated by an EDHF. The results also suggest that ACh may activate the release of ATP from endothelial cells, which in turn activates $P_{2Y}$ receptor on the endothelial cells. Activation of endothelial $P_{2Y}$ receptors induces a release of EDHF resulting in a vasorelaxation via a mechanism that involves activation of both the voltage-gated $K^+$ channels and the $Ca^{2+}-activated\;K^+\;channels$. The results further suggest that EDHF does not appear to be a cytochrome P450 metabolite.

• Korean Journal of Veterinary Research
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• v.34 no.1
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• pp.25-36
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• 1994

The inward tail current after a short depolarizing pulse has been known as Na-Ca exchange current activated by intracellular calcium which forms late plateau of the action potential in rabbit atrial myocytes. Chloride conductance which is also dependent upon calcium concentration has been reported as a possible tail current in many other excitable tissues. Thus, in order to investigate the exsitance of the calcium activated chloride current and its contribution to tail current, whole cell voltage clamp measurement has been made in single atrial cells of the rabbit. The current was recorded during repolarization following a brief 2 ms depolarizing pulse to +40mV from a holding potential of -70mV. When voltage-sensitive transient outward current was blocked by 2 mM 4-aminopyridine or replacement potassium with cesium, the tail current were abolished by ryanodine$(1{\mu}M)$ or diltiazem$(10{\mu}M)$ and turned out to be calcium dependent. The magnitudes of the tail currents were increased when intracellular chloride concentration was increased to 131 mM from 21 mM. The current was decreased by extracellular sodium reduction when intracellular chloride concentration was low(21 mM), but it was little affected by extracellular sodium reduction when intracellual chloride concentration was high(131 mM). The current-voltage relationship of the difference current before and after extracellular sodium reduction, shows an exponential voltage dependence with the largest magnitude of the current occurring at negative potentials, with is similar to current-voltage relationship at negative potentials, which is similar to current-voltage relationship of Na-Ca exchange current. The current was also decreased by $10{\mu}M$ niflumic acid and 1 mM bumetanide, which is well known anion channel blockers. The reversal potentials shifted according to changes in chloride concentration. The current-voltage relationships of the niflumic acid-sensitive currents in high and low concentration of chloride were well fitted to those predicted as chloride current. From the above results, it is concluded that calcium activated chloride component exists in the tail current with Na-Ca exchange current and it shows the reversal of tail current. Therefore it is thought that in the physiologic condition it leads to rapid end of action potential which inhibits calcium influx and it contributes to maintain the low intracellular calcium concentration with Na-Ca exchange mechanism.

• The Korean Journal of Pain
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• v.27 no.3
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• pp.229-238
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• 2014

Background: A toxic dose of bupivacaine produces vasodilation in isolated aortas. The goal of this in vitro study was to investigate the cellular mechanism associated with bupivacaine-induced vasodilation in isolated endothelium-denuded rat aortas precontracted with phenylephrine. Methods: Isolated endothelium-denuded rat aortas were suspended for isometric tension recordings. The effects of nifedipine, verapamil, iberiotoxin, 4-aminopyridine, barium chloride, and glibenclamide on bupivacaine concentration-response curves were assessed in endothelium-denuded aortas precontracted with phenylephrine. The effect of phenylephrine and KCl used for precontraction on bupivacaine-induced concentration-response curves was assessed. The effects of verapamil on phenylephrine concentration-response curves were assessed. The effects of bupivacaine on the intracellular calcium concentration ($[Ca^{2+}]_i$) and tension in aortas precontracted with phenylephrine were measured simultaneously with the acetoxymethyl ester of a fura-2-loaded aortic strip. Results: Pretreatment with potassium channel inhibitors had no effect on bupivacaine-induced relaxation in the endothelium-denuded aortas precontracted with phenylephrine, whereas verapamil or nifedipine attenuated bupivacaine-induced relaxation. The magnitude of the bupivacaine-induced relaxation was enhanced in the 100mM KCl-induced precontracted aortas compared with the phenylephrine-induced precontracted aortas. Verapamil attenuated the phenylephrine-induced contraction. The magnitude of the bupivacaine-induced relaxation was higher than that of the bupivacaine-induced $[Ca^{2+}]_i$ decrease in the aortas precontracted with phenylephrine. Conclusions: Taken together, these results suggest that toxic-dose bupivacaine-induced vasodilation appears to be mediated by decreased calcium sensitization in endothelium-denuded aortas precontracted with phenylephrine. In addition, potassium channel inhibitors had no effect on bupivacaine-induced relaxation. Toxic-dose bupivacaine-induced vasodilation may be partially associated with the inhibitory effect of voltage-operated calcium channels.

• The Korean Journal of Pharmacology
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• v.32 no.3
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• pp.301-309
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• 1996

Since it has been reported that the depolarization-induced NE release is inhibited by activation of presynaptic $A_1-adenosine$ heteroreceptor in hippocampus, a large body of experimental data on the post-receptor mechanism of this process has been accumulated. But, the post-receptor mechanism of presynaptic $A_1-adenosine$ receptor on the NE release has not been clearly elucidated yet. Therefore, it was attempted to clarify the participation of $K^+-channel$ in the post-receptor mechanisms of the $A_1-adenosine$ receptor-mediated control of NE release in this study. Slices from rat hippocampus were equilibrated with $^3H-norepinephrine$ and the release of the labelled products was evoked by electrical stimulation (3 Hz, 5 $VCm^{-1}$, 2 ms, rectangular pulses), and the influence of various agents on the evoked tritium-outflow was investigated. Adenosine, in concentrations ranging from $1{\sim}30\;{\mu}M$, decreased the NE release in a dose-dependent manner, without affecting the basal rate of release. 4AP $(1{\sim}30{\mu}M)$, a specific A-type $K^+-channel$ blocker, increased the evoked NE release in a dose-related fashion, and the basal rate of release is increased by 10 and $30{\mu}M$. TEA $(1{\sim}10{\mu}\;M)$, a nonspecific $K^+-channel$ blocker, increased the evoked NE release in a dose-dependent manner without affecting basal release. The adenosine effects were significantly inhibites by 3 ${\mu}M$ 4AP and 10 mM TEA treatment. 4AP $(30{\mu}M)-$ and TEA (10 mM)-induced increments of evoked NE release were completely abolished in $Ca^{++} free, but these were recoverd in low $Ca^{++} medium. And the effects of $K^+-channel$ blockers in low $Ca^{++} medium were inhibites and abolishes by $Mg^{++} (4 mM) adding and TTX $(0.3{\mu}M)$ adding medium, respectively. These results suggest that the decrement of the evoked NE-release by $A_1-adenosine$ receptor is mediated by 4AP and TEA sensitive $K^+-channel$.