• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 1997년도 학술발표회
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• pp.17-17
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• 1997

We investigated the relationship between the voltage-operated calcium channel current and the corresponding [Ca$^{2＋}$]i change (Ca$^{2＋}$-transient) in guinea-pig gastric myocyte. Fluorescence microspectroscopy was combined with conventional whole-cell patch clamp technique and fura-2 (80 $\mu$M) was added into the CsCl-rich pipette solution.(omitted)

• The Korean Journal of Physiology and Pharmacology
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• 제3권6호
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• pp.623-630
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• 1999

Decreased cardiac contractility occurs in endotoxicosis, but little is known about the ionic mechanism responsible for myocardial dysfunction. In this study, we examined the changes in $Ca{2+}$ and $K^+$ currents in cardiac myocytes from endotoxin-treated rat. Ventricular myocytes were isolated from normal and endotoxemic rats (ex vivo), that were treated for 10 hours with Salmonella enteritidis lipopolysaccharides (LPS; 1.5 mg/kg) intravenously. Normal cardiac myocytes were also incubated for 6 hours with 200 ng/ml LPS (in vitro). L-type $Ca{2+}$ current $(I_{Ca,L})$ and transient outward $K^+$ current $(I_{to})$ were measured using whole cell patch clamp techniques. Peak $I_{Ca,L}$ was reduced in endotoxemic myocytes (ex vivo; 6.00.4 pA/pF, P＜0.01) compared to normal myocytes (control; 10.90.6 pA/pF). Exposure to endotoxin in vitro also attenuated $I_{Ca,L}$ (8.40.4 pA/pF, P＜0.01). The amplitude of $(I_{to})$ on depolarization to 60 mV was reduced in endotoxin treated myocytes (16.51.5 pA/pF, P＜0.01, ex vivo; 20.00.9 pA/pF, P＜0.01 , in vitro) compared to normal myocytes (control; 24.71.0 pA/pF). There was no voltage shift in steady-state inactivation of $I_{Ca,L}$ and $(I_{to})$ between groups. These results suggest that endotoxin reduces $Ca{2+}$ and $K^+$ currents of rat cardiac myocytes, which may lead to cardiac dysfunction.

• The Korean Journal of Physiology and Pharmacology
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• 제23권3호
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• pp.219-227
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• 2019

Polycystic kidney disease 2-like-1 (PKD2L1), polycystin-L or transient receptor potential polycystin 3 (TRPP3) is a TRP superfamily member. It is a calcium-permeable non-selective cation channel that regulates intracellular calcium concentration and thereby calcium signaling. Although the calmodulin (CaM) inhibitor, calmidazolium, is an activator of the PKD2L1 channel, the activating mechanism remains unclear. The purpose of this study is to clarify whether CaM takes part in the regulation of the PKD2L1 channel, and if so, how. With patch clamp techniques, we observed the current amplitudes of PKD2L1 significantly reduced when co-expressed with CaM and $CaM{\triangle}N$. This result suggests that the N-lobe of CaM carries a more crucial role in regulating PKD2L1 and guides us into our next question on the different functions of two lobes of CaM. We also identified the predicted CaM binding site, and generated deletion and truncation mutants. The mutants showed significant reduction in currents losing PKD2L1 current-voltage curve, suggesting that the C-terminal region from 590 to 600 is crucial for maintaining the functionality of the PKD2L1 channel. With PKD2L1608Stop mutant showing increased current amplitudes, we further examined the functional importance of EF-hand domain. Along with co-expression of CaM, ${\triangle}EF$-hand mutant also showed significant changes in current amplitudes and potentiation time. Our findings suggest that there is a constitutive inhibition of EF-hand and binding of CaM C-lobe on the channel in low calcium concentration. At higher calcium concentration, calcium ions occupy the N-lobe as well as the EF-hand domain, allowing the two to compete to bind to the channel.

• 대한한의학회지
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• 제19권2호
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• pp.450-467
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• 1998

To research the characteristics of ion currents induced by inhibitory and excitatory herbs of oriental medicine, we used nystatin-perforated patch clamp technique under voltage clamp condition in periaqueductal gray neuron dissociated from Sprauge-Dawley rat, 10-15 days old. The results are as follows. 1. Ion current induced by $10mg/m{\ell}$ of Bupleuri Radix was inhibited $59.50{\pm}4.29%$ by $10^{-4}M$ bicuculline(p>0.01) but inhibition of $10.75{\pm}4.77%$ by $10^{-4}M$ tubocurarine and $4.75{\pm}4.23%$ by $10^{-4}M$ verapamil had no statistical significance(p>0.05). So ion current induced by Bupleuri Radix revealed only GABA induced $Cl^-$ current, not acetylcholine and $Ca^{2+}$ current. 2. Ion current induced by $20mg/m{\ell}$ of Coptidis Rhizoma was inhibited $47.20{\pm}7.88%$ by $10^{-4}M$ bicuculline(p<0.01) but $3.20{\pm}2.33%$ inhibition by $10^{-4}M$ tubocurarine and $1.00{\pm}1.00%$ inhibition by $10^{-4}M$ verapamil had no significance(p>0.05). So ion current induced by Coptidis Rhizoma revealed only GABA induced $Cl^-$ current, not acetylcholine and $Ca^{2+}$ current. 3. Ion current induced by $20mg/m{\ell}$ of Ecliptae Herba was inhibited $55.00{\pm}4.92%$ by $10^{-4}M$ bicuculline (p<0.01), and also inhibited $15.00{\pm}4.26%$ by $10^{-4}M$ tubocurarine(p<0.05), but inhibition of $6.00{\pm}3.03%$ by $10^{-4}M$ verapamil had no significance(p>0.05). So ion current induced by Ecliptae Herba showed GABA activated $Cl^-$ current and acetylcholine activated cation current, not $Ca^{2+}$ current 4. Ion current induced by $5mg/m{\ell}$ of Liriopis Tuber was inhibited $15.20{\pm}4.57%$ by $10^{-4}M$ bicuculline<0.05) and also inhibited $14.00{\pm}3.00%$ by $10^{-4}M$ tubocurarine(p<0.05), but inhibition of $5.20{\pm}4.80%$ by $10^{-4}M$ verapamil had no significance(p>0.05). So ion current induced by Liriopis Tuber showed GABA. activated $Cl^-$ current and acetylcholine activated cation current, not $Ca^{2+}$ current. 5. Ion current induced by $5mg/m{\ell}$ of Aconiti Tuber was inhibited $97.00{\pm}1.34%$ by $10^{-4}M$ bicuculline(p<0.01), $80.00{\pm}9.83%$ by $10^{-4}M$ tubocurarine(p<0.01), and $24.00{\pm}6.18%$ by $10^{-4}M$ verapamil(p<0.05). So ion current induced by Aconiti Tuber revealed GABA activated $Cl^-$ current and acetylcholine activated cation current and $Ca^{2+}$ current. 6. Ion current induced by $10mg/m{\ell}$ of Zingiberis Rhizoma was inhibited $33.00{\pm}7.43%$ by $10^{-4}$ bicuculline(p<0.05), $10.20{\pm}1.83%$ by $10-^{-4}M$ tubocurarine(p<0.01), and $14.00{\pm}2.16%$ by $10^{-4}M$ verapamil(p<0.01) So ion current induced by Zingiberis Rhizoma revealed GABA activated $Cl^-$ current and acetylcholine activated cation outtent and $Ca^{2+}$ current. 7. Ion current induced by $10mg/m{\ell}$ of Boshniakiae Herba was inhibited $65.00{\pm}13.75%$ by $10^{-4}M$ bicuculline(p<0.05), $38.00{\pm}9.24%$ by $10^{-4}M$ tubocurarine(p<0.05), and $33.25{\pm}7.42%$ by $10^{-4}M$ verapamiHp<0.05). So ion current induced by Bpshniakiae Herba revealed GABA activated $Cl^-$ current and acetylcholine activated cation current and $Ca^{2+}$ current. These results suggest that a point of difference between inhibitory and excitatory herbs is existence of$Ca^{2+}$ current.

• 한국수정란이식학회지
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• 제16권2호
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• pp.79-89
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• 2001

배분화과정시 나타나는 $Ca^{2+}$ 변화에 미치는 $E_2$의 영향을 알아보고자 whole cell voltage clamp 기법, 방사선 등위원소 면역측정법, 그리고 공초점 현미경을 통하여 $E_2$처리 후 나타나는 $Ca^{2+}$ 전류 변화 및 세포내 $Ca^{2+}$ 농도 변화를 조사하였다. 생쥐의 미성숙 난자는 난소의 난포를 천자하고, 배란난자는 과배란 처리 후 난관에서 회수하였다. 수정란은 과배란 처리 후 수컷 생쥐와 교미를 유도한 후 각각의 단계에 맞는 수정란을 채란하였다. 혈중 $E_2$의 농도는 심장을 천자하여 혈액을 채취한 후 배발달 단계와 호르몬 처리 시간이 일치하는 혈액만을 사용하였다. 본 실험의 결과를 요약하면 다음과 같다. 1. $E_2$처리시 미성숙난자의 제 1극체 형성률 (성숙의 지표)은 $E_2$를 처리하지 않은 난자(83% : 83/100)보다 $E_2$를 처리한 난자 (94%, 94/100)에서 유의적 (P<0.05)으로 높게 나타났다. 2. $E_2$를 처리하였을 때 $Ca^{2+}$ 내향전류의 변화는 -10 mV에서 -1.23$\pm$0.01 nA (n=15)에서 -1.50$\pm$0.03 nA (n=15)로 122% 상승함으로써 유의한 (P<0.05) 변화를 보였다. 3. $E_2$를 처리하지 않은 난자 및 수정란을 1로 한 후 $E_2$를 처리한 난자 및 수정란의 변화를 상대적인 값으로 표시하였다. $E_2$처리한 난자는 1.22$\pm$0.17 (n=10), $E_2$처리한 전핵배는 1.20$\pm$0.14 (n=10), $E_2$처리한 2세포기배는 1.07$\pm$0.01 (n=10), 4세포기배는 1.05$\pm$0.09 (n=10)를 나타냄으로써 수정란의 단계마다 $E_2$의 반응 결과가 차이가 남을 알 수 있었다. 4. $E_2$농도 곡선에서 PMSG 처리 후 $E_2$의 혈중농도는 계속적인 상승을 보이다가 배란시기에 최고치를 나타내었으며, 배란 후 다시 감소하여 8세포기에서는 급격한 감소현상이 나타났다. 이후 다시 상실기를 거쳐 배반포기 임신기간동안 $E_2$의 농도가 상승하였다. 5. $E_2$처리 후 세포내 $Ca^{2+}$ 농도변화의 결과로, $E_2$를 처리하지 않은 난자들의 세포내 $Ca^{2+}$ 농도는 836.4$\pm$131.2 (n=10), $E_2$를 처리한 난자들은 1736.4$\pm$192.0 (n=10)로써 유의한 (P<0.05) 차이를 보였다. 이상의 결과로부터 $E_2$처리에 의한 세포내 $Ca^{2+}$ 농도 상승은 $E_2$가 $Ca^{2+}$ 통로를 자극함으로써 세포바깥의 $Ca^{2+}$이 세포안으로 이동하여 나타나는 변화로 생각된다.

• 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.

• Journal of Welding and Joining
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• 제12권4호
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• pp.76-84
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• 1994

Droplet transfer modes due to welding conditions and the effect of Ca in welding wire on droplet transfer were investigated. Droplet transfer mode in CO$_{2}$ welding was classified into 2 modes, that is, short circuit and globular transfer, with increasing welding current and voltage. With increasing Ca content in wire, repulsive pressure due to vaporization of Ca was considerably increased. In short circuit transfer region, arcing time was increased and droplet transfer cycle was decreased, with increasing Ca content. In globular transfer region, welding condition for globular transfer was lower current region, with increasing Ca content.

• The Korean Journal of Physiology
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• 제23권2호
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• pp.313-328
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• 1989

The electrophysiological properties of the inward current contributing to the late plateau phase of the action potential were investigated using the whole cell clamp technique and intracellular dialysis in single atrial cells isolated from the rabbit heart. The inward current was activated by various repolarizing pulses after a brief depolarizing pulse to +40 mV for 2 ms and its time course was similar to that of the late plateau of the action potential. The current was fully activated above the potential of -40 mV and abolished by intracellular EGTA. Ryanodine of $1{\mu}M$ also abolished the late plateau and the inward current. Reduced $Na_o\;to\;30%\;and\;20\;mM\;Na_1$ diminished the late plateau together with the inward current. Diltiazem blocked the activation of the current and Ni in the concentration of $40{\sim}200\;{\mu}M$ decreased the development of the late plateau and the inward current. Fully activated current-voltage relation of the inward current showed exponential voltage dependency which was steeper in more hyperplarizing range. The above findings suggest that the inward current was activated by intracellular calcium and contribute the late plateau phase of the action potential. It could be concluded that the inward current would be the inward component of Na-Ca exchange.

• 공업화학
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• 제23권5호
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• pp.474-479
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• 2012

막결합 축전식 탈염(MCDI) 기술을 이용하여 $Na^{+}$와 $Ca^{2+}$ 이온이 혼합된 용액에서 $Ca^{2+}$ 이온의 선택적 제거 가능성을 연구하였다. 양이온교환막인 CMX막에 대한 $Ca^{2+}$ 이온의 선택성을 확인하기 위해 흡착평형 실험을 실시하였다. 그리고 MCDI 셀을 이용해 혼합용액(5 meq/L NaCl + 2 meq/L $CaCl_{2}$)에 대한 탈염실험을 수행하였다. 흡착평형 실험결과 용액과 CMX막에서 $Ca^{2+}$ 이온의 당량분율은 각각 28.6, 87.2%를 보여 CMX막이 $Ca^{2+}$ 이온에 대해 높은 선택성을 갖는 것을 확인하였다. MCDI 셀에 일정한 전류를 인가하면서 셀 전위가 1.0 V에 도달할 때까지 탈염실험을 실시하였다. 그 결과 흡착된 이온의 총량은 인가한 전류밀도에 큰 영향을 받지 않고 일정하였다. 그러나 흡착된 이온 중 $Ca^{2+}$ 이온의 비율은 전류밀도에 반비례하였으며 200, 300, 500, $700\;A/m^{2}$의 전류밀도에서 각각 81.4, 78.4, 77.0, 74.5%로 나타났다. 이러한 결과는 낮은 전류밀도에서 CMX막에 흡착된 $Ca^{2+}$ 이온의 비율이 높았기 때문인 것으로 판단된다.

• The Korean Journal of Physiology and Pharmacology
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• 제21권2호
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• pp.241-249
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• 2017

Plasma membrane hyperpolarization associated with activation of $Ca^{2+}$-activated $K^+$ channels plays an important role in sperm capacitation during fertilization. Although Slo3 (slowpoke homologue 3), together with the auxiliary ${\gamma}^2$-subunit, LRRC52 (leucine-rich-repeat-containing 52), is known to mediate the pH-sensitive, sperm-specific $K^+$ current KSper in mice, the molecular identity of this channel in human sperm remains controversial. In this study, we tested the classical $BK_{Ca}$ activators, NS1619 and LDD175, on human Slo3, heterologously expressed in HEK293 cells together with its functional interacting ${\gamma}^2$ subunit, hLRRC52. As previously reported, Slo3 $K^+$ current was unaffected by iberiotoxin or 4-aminopyridine, but was inhibited by ~50% by 20 mM TEA. Extracellular alkalinization potentiated hSlo3 $K^+$ current, and internal alkalinization and $Ca^{2+}$ elevation induced a leftward shift its activation voltage. NS1619, which acts intracellularly to modulate hSlo1 gating, attenuated hSlo3 $K^+$ currents, whereas LDD175 increased this current and induced membrane potential hyperpolarization. LDD175-induced potentiation was not associated with a change in the half-activation voltage at different intracellular pHs (pH 7.3 and pH 8.0) in the absence of intracellular $Ca^{2+}$. In contrast, elevation of intracellular $Ca^{2+}$ dramatically enhanced the LDD175-induced leftward shift in the half-activation potential of hSlo3. Therefore, the mechanism of action does not involve pH-dependent modulation of hSlo3 gating; instead, LDD175 may modulate $Ca^{2+}$-dependent activation of hSlo3. Thus, LDD175 potentially activates native KSper and may induce membrane hyperpolarization-associated hyperactivation in human sperm.