• The Korean Journal of Physiology and Pharmacology
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• 제2권2호
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• pp.225-232
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• 1998

To investigate the possible involvement of outward potassium ($K^+$) currents in nitric oxide-induced relaxation in intestinal smooth muscle, we used whole-cell patch clamp technique in freshly dispersed guinea-pig ileum longitudinal smooth muscle cells. When cells were held at -60 mV and depolarized from -40 mV to -50 mV in 10 mV increments, sustained outward $K^+$ currents were evoked. The outward $K^+$ currents were markedly increased by the addition of 10 ${\mu}M$ sodium nitroprusside (SNP). 10 ${\mu}M$ S-nitroso-N-acetylpenicillamine (SNAP) and 1 mM 8-Bromo-cyclic GMP (8-Br-cGMP) also showed a similar effect to that of SNP. 1 mM tetraethylammonium (TEA) significantly reduced depolarization-activated outward $K^+$ currents. SNP-enhanced outward $K^+$ currents were blocked by the application of TEA. High EGTA containing pipette solution (10 mM) reduced the control currents and also inhibited the SNP-enhanced outward $K^+$ currents. 5 mM 4-aminopyridine (4-AP) significantly reduced the control currents but showed no effect on SNP-enhanced outward $K^+$ currents. 0.3 ${\mu}M$ apamin and 10 ${\mu}M$ glibenclamide showed no effect on SNP-enhanced outward $K^+$ currents. 10 ${\mu}M$ 1H-[1,2,4]oxadiazolo [4,3-a]quinoxaline-1-one (ODQ), a specific inhibitor of soluble guanylate cyclase, significantly blocked SNP-enhanced $K^+$ currents. We conclude that NO donors activate the $Ca^{2+}-activated$ $K^+$ channels in guinea-pig ileal smooth muscle via activation of guanylate cyclase.

• The Korean Journal of Physiology and Pharmacology
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• 제1권2호
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• pp.169-183
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• 1997

We have investigated the two types of voltage-dependent outward potassium (K) currents, i.e. delayed rectifier K current ($I_{K(V)}$) and 'A-like' transient outward K current ($I_{to}$) with patch-clamp technique in single smooth muscle cells (SMCs) isolated from rabbit basilar artery, and investigated the characteristics of them. The time-courses of activation were well fitted by exponential function raised to second power ($n^2$) in $I_{K(V)}$ and fourth power ($n^4$) in $I_{to}$. The activation, inactivation and recovery time courses of $I_{to}$ were much faster than that of $I_{K(V)}$. The steady-state activation and inactivation of $I_{K(V)}$ was at the more hyperpolarized range than that of $I_{to}$ contrary to the reports in other vascular SMCs. Tetraethylammonium chloride (TEA; 10 mM) markedly inhibited $I_{K(V)}$ but little affected $I_{to}$. 4-Aminopyridine (4-AP) had similar inhibitory potency on both currents. While a low concentration of $Cd^{2+}$ (0.5 mM) shifted the current- voltage relationship of $I_{to}$ to the positive direction without change of maximum conductance, $Cd^{2+}$ did not cause any appreciable change for $I_{K(V)}$.

• 대한화학회지
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• 제53권3호
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• pp.308-324
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• 2009

현재 수행하는 작업은 피페리딘 또는 암모늄 아세테이트 존재하에서 malononitrile와 $\beta$- aroylacrylic acid 유도체의 DMF 용매조건에서 상호작용에 대한 연구이며, 형성된 화합물을 이용한 퓨즈 되고 단리된 이중고리화 시스템의 합성에 관한 것이다. $\beta$-aroylacrylic acid (3)이 DMF 용매와 피페리딘 촉매조건에malononitrile와 반응하여 4H-피란유도체(4)를 형성한다. 촉매를 암모늄아세테이트로 바꿈으로서 피리딘 유도체를 얻었다. 또한 N-말레암산 유도체 (19)와 (27)은 말레 무수물과 함께 (4)와 (5)의 반응을 경유하여 합성되었다. 마이클 첨가 반응에서 이용되는 메틸렌화합물에 관한 이 연구는 B-aroylacrylic acid의 경우와 유사하게 형성된 말레암산 유도체의 반응성에 대한 것이다.

• The Korean Journal of Physiology and Pharmacology
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• 제12권2호
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• pp.73-77
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• 2008

Recent studies have documented that testosterone relaxes several smooth muscles by modulating $K^+$ channel activities. Smooth muscles of seminal vesicles playa fundamental role in ejaculation, which might involve testosterone. This study was aimed to assess the role of testosterone in seminal vesicular motility by studying its effects on contractile agents and on the ion channels of single vesicular myocytes in a rabbit model. The contractile responses of circular smooth muscle strips of rabbit seminal vesicles to norepinephrine ($10{\mu}M$), a high concentration of KCI (70 mM), and testosterone ($10{\mu}M$) were observed. Single vesicular myocytes of rabbit were isolated using proteolytic enzymes including collagenase and papain. Inside-out, attached, and whole-cell configurations were examined using the patch clamp technique. The applications of $10{\mu}M$ norepinephrine or 70 mM KCl induced tonic contractions, and $10{\mu}M$ testosterone (pharmacological concentration) evoked dose-dependent relaxations of these precontracted strips. Various $K^+$ channel blockers, such as tetraethylammonium (TEA; $10{\mu}M$), iberiotoxin ($0.1{\mu}M$), 4-aminopyridine (4-AP, $10{\mu}M$), or glibenclamide ($10{\mu}M$) rarely affected these relaxations. Single channel data (of inside-out and attached configurations) of BK channel activity were also hardly affected by testosterone ($10{\mu}M$). On the other hand, however, testosterone reduced L-type $Ca^{2+}$ currents significantly, and found to induce acute relaxation of seminal vesicular smooth muscle and this was mediated, at least in part, by $Ca^{2+}$ current inhibition in rabbit.

• 대한화학회지
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• 제30권2호
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• pp.145-151
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• 1986

3-CN, 4-CN, $3-NH_2$ 및 $4-NH_2$ 치환피리딘의 해리상수를 온도(15 ∼ 40${\circ}C$)와 압력(1-2500bar)을 변화시키면서 측정하였다. 측정방법은 전도도법을 적용했으나 낮은 해리상수를 가진 물질에서도 적용되는 피리딘류의 염용액의 전도도를 측정하므로 계산가능한 방법을 모색하여 응용하였다. 이 방법은 어떤 조건에서의 해리상수를 알고 있어야 하는 것과 실험값을 외삽하여 피리딘염의 전도도를 알아야 하는 번거로움도 있으나 측정이 가능하다는 것을 확인할 수 있다. 이들 치환체의 해리상수는 온도가 증가하면 커지고 압력이 높아지면 해리상수는 작아진다는 것을 알 수 있었다. 또 반응의 열역학적 성질을 계산하여 반응성도 고찰하였다.

• Animal cells and systems
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• 제1권1호
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• pp.81-86
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• 1997

We studied the mechanism of arachidonic acid on the secretion of a neurotransmitter in rat pheochromocytoma PC12 cells. Arachidonic acid inhibited the 70 mM $K^+$-induced secretion of norepinephrine. Arachidonic acid also inhibited the 70 mM $K^+$-induced $Ca^{2+}$ mobilization which is due to the opening of the voltage-sensitive $Ca^{2+}$ channels (VSCC). Both the half maximal inhibitory concentration ($IC_{50}$) of the norepinephrine secretion and VSCC coincided at 30 uM. The major oxidized metabolites of arachidonic acid, prostaglandins did not mimic the inhibitory effect of arachidonic acid. Nordihydroguaiaretic acid (NDGA) and indomethacin which are inhibitors of lipoxygenase and cyclooxygenase, respectively, did not block the inhibitory effect of arachidonic acid. The results suggest that arachidonic acid serves as a signal itself, not in the form of metabolites. The pretreatment of various $K^+$ channel blockers such as 4-aminopyridine, tetraethylarnmonium, glipizide, or glibenclamide also did not show any effect on the inhibitory effect of arachidonic acid. Through these results we suggest that arachidonic acid regulates VSCC directly and affects the secretion of neurotransmitters.

• 대한약리학회지
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• 제27권2호
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• pp.99-108
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• 1991

Racemic ketamine을 사용하여 개구리의 좌골신경 및 toe muscle에 대한 작용을 관찰하였다. 실험방법으로는 214 mM sucrose을 사용하여 서로 다른 두 종류의 투여 방법으로 세포막의 활동 전압에 대한 영향을 electric recording으로 관찰하였다. 즉, intracellular 투여는 single sucrose gap technique으로, extracellar 투여는 double sucrose gap technique을 사용하였으며 그 실험 결과는 아래와 같았다. 1. Racemic ketamine은 개구리의 좌골신경 및 toe muscle의 활동전압을 intracellular 및 extracellular 투여시 모두 의의 있게 억제하였다. 2. 개구의 toe muscle에서 $K^+$-수축을 억제하였다. 3. naloxone은 ketamine의 억제작용을 완전히 차단하지는 못하였다.

• The Korean Journal of Physiology
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• 제26권1호
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• pp.27-35
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• 1992

We used the whole cell patch clamp technique to examine the ionic basis for the tail current after depolarizing pulse in single atrial myocytes of the rabbit. We recorded the tail currents during various repolarizations after short depolarizing pulse from a holding potential of -70 mV. The potassium currents were blocked by external 4-aminopyridine and replacement of internal potassium with cesium. The current was reversed to the outward direction above +10 mV. High concentrations of intracellular calcium buffer inhibited the activation of the current. Diltiazem and ryanodine blocked it too. These data suggest that the current is activated by intracellular calcium released from sarcoplasmic reticulumn. When the internal chloride concentration was increased, the inward tail current was increased. The current was partially blocked by the anion transport blocker niflumic acid. The current voltage curve of the niflumic acid sensitive current component shows outward rectification and is well fitted to the current voltage curve of the theoretically predicted chloride current calculated from the constant field equation. The currents recorded in rabbit atrial myocytes, with the method showing isolated outward Na Ca exchange current in ventricular cells of the guinea pig, suggested that chloride conductance could be activated with the activation of Na/ca exchange current. From the above results it is concluded that a chloride sensitive component which is activated by intracellular calcium contributes to tail currents in rabbit atrial cells.

• 약학회지
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• 제41권4호
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• pp.399-406
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• 1997

The effects of different $K^+$ channel blockers were investigated on the non-adrenergic non-cholinergic (NANC) relaxations in the circular muscle of the rabbit proximal stomach. Non-selective blockers of $K^+$ channels, 4-aminopyridine (4-AP, 3~30${\mu}M$) and tetraethylammonium (TEA, 100~1000${\mu}M$) significantly enhanced the NANC relaxations in a concentration-dependent manner. The enhancement was more prominent for the NANC relaxations induced by the electric field stimulation (EFS) with lower frequencies. Blockers of large conductance $Ca^{2+}$-activated $K^+$ channels, charybdotoxin and iberiotoxin, a blocker of small conduntance $Ca^{2+}$-activated $K^+$ channels, apamin and a blocker of ATP-sensitive $K^+$ channels, glibenclamide had no effect on the NANC relaxations, respectively. Exogeneous administration of nitric oxide (NO, 1~30${\mu}M$) caused concentration-dependent relaxations which showed a similarity to those obtained with EFS. None of the $K^+$ channel blockers had an effect on the concentration-dependent relaxation in response to NO. These results suggest that prejunctional $K^+$ channels regulate the release of NO from the NANC nerve in the rabbit proximal stomach as the inhibition of prejunctional $K^+$ channels increases the NANC relaxation induced by the EFS.

• The Korean Journal of Physiology and Pharmacology
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• 제12권4호
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• pp.131-135
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• 2008

The profile of membrane currents was investigated in differentiated neuronal cells derived from human neural stem cells (hNSCs) that were obtained from aborted fetal cortex. Whole-cell voltage clamp recording revealed at least 4 different currents: a tetrodotoxin (TTX)-sensitive $Na^+$ current, a hyperpolarization-activated inward current, and A-type and delayed rectifier-type $K^+$ outward currents. Both types of $K^+$ outward currents were blocked by either 5 mM tetraethylammonium (TEA) or 5 mM 4-aminopyridine (4-AP). The hyperpolarization-activated current resembled the classical $K^+$ inward current in that it exhibited a voltage-dependent block in the presence of external $Ba^{2+}$ (30 ${\mu}$M) or $Cs^+$ (3${\mu}$M). However, the reversal potentials did not match well with the predicted $K^+$ equilibrium potentials, suggesting that it was not a classical $K^+$ inward rectifier current. The other $Na^+$ inward current resembled the classical $Na^+$ current observed in pharmacological studies. The expression of these channels may contribute to generation and repolarization of action potential and might be regarded as functional markers for hNSCs-derived neurons.