• 제목/요약/키워드: patch-clamp technique

검색결과 155건 처리시간 0.028초

Effects of Ethanol on GABA-Activated Chloride Current in Sprague-Dawley rat Hippocampal Neurons

  • Sohn, Yeong-Jae;Chung, In-Kyo;Kim, Inn-Se;Cho, Goon-Jae;Chung, Yong-Za;Il Yun
    • Journal of Life Science
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    • 제9권2호
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    • pp.15-18
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    • 1999
  • Tempting to further understand the molecular mechanism of pharmacological action of ethanol, we investigated the acute effects of ethanol on the GABA-activated current (IGABA) of the cultured Sprague-Dawley rat hippocampal neurons in primary culture using the whole-cell patch-clamp technique. Patch-clamp recordings revealed that ethanol potentiated the Cl- current in a concentration-dependent manner(1-300mM) in the majority of the cell studied. This study demonstrates that ethanol can potentiate IGABA in mammalian central neurons.

Effects of Samchulkunbi-tang in Cultured Interstitial Cells of Cajal of Murine Small Intestine

  • Kim, Jung Nam;Kwon, Young Kyu;Kim, Byung Joo
    • 동의생리병리학회지
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    • 제27권1호
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    • pp.112-117
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    • 2013
  • We studied the modulation of pacemaker activities by Samchulkunbi-tang (SCKB) in cultured interstitial cells of Cajal (ICC) from murine small intestine with the whole-cell patch-clamp technique. Externally applied SCKB produced membrane depolarization in the current-clamp mode. The pretreatment with $Ca^{2+}$-free solution and thapsigargin, a $Ca^{2+}$-ATPase inhibitor in endoplasmic reticulum, abolished the generation of pacemaker potentials and suppressed the SCKB-induced action. The application of flufenamic acid (a nonselective cation channel blocker) abolished the generation of pacemaker potentials by SCKB. However, the application of niflumic acid (a chloride channel blocker) did not inhibit the generation of pacemaker potentials by SCKB. In addition, the membrane depolarizations were inhibited by not only GDP-${\beta}$-S, which permanently binds G-binding proteins, but also U-73122, an active phospholipase C inhibitor. These results suggest that SCKB modulates the pacemaker activities by nonselective cation channels and external $Ca^{2+}$ influx and internal $Ca^{2+}$ release via G-protein and phospholipase C-dependent mechanism. Therefore, the ICC are targets for SCKB and their interaction can affect intestinal motility.

The Effect of Ethanol on 5-Hydrosytryptamine Receptor-Mediated Ion Current in Cultured NCB-20 Neuroblastoma Cells

  • Woo, Hyo-Geyng;Chung, In-Kyo;Cho, Goon-Jae;Chung, Yong-Za;Il Yun
    • Journal of Life Science
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    • 제9권2호
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    • pp.82-85
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    • 1999
  • The effects of ethanol on 5-hydrosytryptamine(5-HT3) receptor-mediated ion current were evaluated in whole-cell patch-clamp recordings from NCB-20 neuroblastoma cells. The physiologic and pharmacologic properties of 5-HT-activated ion current in NCB-20 cells indicated that it was mediated by 5-HT3 receptors. Ethanol(25-100mM) potentiated 5-HT3 receptor-mediated current in a concentration-dependent manner.

천수근이 중심회백질 신경세포에서 이온통로 조절작용에 미치는 영향 (Modulation of Harpagophytum procumbens on ion channels in acutely dissociated periaqueductal gray neurons of rats)

  • 신민철;장현경;장미현;김창주;김연희;김이화
    • Korean Journal of Acupuncture
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    • 제20권4호
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    • pp.17-29
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    • 2003
  • 목적:천수근은 염증성 관절염과 염증성 장 질환 등에 사용되어왔다. 천수근이 중심회백질 신경세포에서 하행성 진통작용에 미치는 영향을 연구하였다. 방법: 중심회백질 신경세포를 분리하여 전위고정하에서 nystatin-perforated patch-clamp technique을 시행하였다. 결과: 천수근에 의하여 유발되는 이온전류는 GABA, glycine, 그리고 glutamate 수용체를 모두 활성화시켰다.

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[$Ca^{2+}-activated\;Cl^-$ Current in Gastric Antral Myocytes

  • Lee, Moo-Yeol;Bang, Hyo-Weon;Uhm, Dae-Yong;Rhee, Sang-Don
    • The Korean Journal of Physiology
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    • 제28권2호
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    • pp.143-150
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    • 1994
  • The whole-cell mode of the patch clamp technique was used to study $Ca^{2+}-activated\;Cl^-\;current$ $(I_{Cl_{Ca}})$ in gastric antral myocytes. Extracellular application of caffeine evoked $Ca^{2+}-activated\;current$. In order to isolate the chloride current from background current, all known systems were blocked with specific blockers. The current-voltage relationship of caffeine-induced current showed outward rectification and it reversed at around $E_{Cl^-}$. The shift of reversal potential upon the alteration of external and internal chloride concentrations was well fitted with results which were calculated by the Nernst equation. Extracellular addition of N-phenylanthranilic acid and niflumic acid which are known anion channel blockers abolished the caffeine induced current. Intracellular application of a high concentration of EGTA also abolished this current. Application of c-AMP, c-GMP, heparin, or $AIF^-_4$ made no remarkable changes to this current. Sodium replacement with the impermeable cation N-methylglucamine or with $Cd^{2+}$ rarely affected this current. From the above results it is suggested that the caffeine induced current was a $Cl^-$ current and it was activated by intracellular $Ca^{2+}$.

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The Alteration of $Ca^{2+}-activated\;K^+$ Channels in Coronary Arterial Smooth Muscle Cells Isolated from Isoproterenol-induced Cardiac Hypertrophy in Rabbit

  • Kim, Na-Ri;Han, Jin;Kim, Eui-Yong
    • The Korean Journal of Physiology and Pharmacology
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    • 제5권2호
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    • pp.147-156
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    • 2001
  • It has been proposed that $Ca^{2+}-activated$ K $(K_{Ca})$ channels play an essential role in vascular tone. The alterations of the properties of coronary $K_{Ca}$ channels have not been studied as a possible mechanism for impaired coronary reserve in cardiac hypertrophy. The present studies were carried out to determine the properties of coronary $K_{Ca}$ channels in normal and hypertrophied hearts. These channels were measured from rabbit coronary smooth muscle cells using a patch clamp technique. The main findings of the present study are as follows: (1) the unitary current amplitudes and the slope conductance of coronary $K_{Ca}$ channels were decreased without changes of the channel kinetics in isoproterenol-induced cardiac hypertrophy; (2) the sensitivity of coronary $K_{Ca}$ channels to the changes of intracellular concentration of $Ca^{2+}$ was reduced in isoproterenol-induced cardiac hypertrophy. From above results, we suggest for the first time that the alteration of $K_{Ca}$ channels are involved in impaired coronary reserve in isoproterenol-induced cardiac hypertrophy.

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Effects of Prostaglandin $F_{2{\alpha}}$ on Membrane Potentials and $K^+$ Currents in Rabbit Middle Cerebral Arterial Cells

  • Kim, Na-Ri;Han, Jin;Kim, Won-Gue;Kim, Eui-Yong
    • The Korean Journal of Physiology and Pharmacology
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    • 제4권4호
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    • pp.301-309
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    • 2000
  • The purpose of our investigation was to examine the effects of prostaglandin $F_{2{\alpha}}\;(PGF_{2{\alpha}})$ on membrane potentials, $Ca^{2+}-activated\;K^+\;(K_{Ca})$ channels, and delayed rectifier $K^+(K_V)$ channels using the patch-clamp technique in single rabbit middle cerebral arterial smooth muscle cells. $PGF_{2{\alpha}}$ significantly hyperpolarized membrane potentials and increased outward whole-cell K currents. $PGF_{2{\alpha}}$ increased open-state probability of $K_{Ca}$ channels without the change of the open and closed kinetics. $PGF_{2{\alpha}}$ increased the amplitudes of $K_V$ currents with a leftward shift of activation and inactivation curves and a decrease of activation time constant. Our results suggest that the activation of $K_{Ca}$ and $K_V$ channels, at least in part, may lead to attenuate or counteract vasoconstriction by $PGF_{2{\alpha}}$ in middle cerebral artery.

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The Effects of Intracellular Monocarboxylates on the ATP-sensitive Potassium Channels in Rabbit Ventricular Myocytes

  • Kim, Na-Ri;Han, Jin;Kim, Eui-Yong;Ho, Won-Kyung;Earm, Yung E.
    • The Korean Journal of Physiology and Pharmacology
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    • 제2권5호
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    • pp.581-589
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    • 1998
  • A regulating mechanism of the ATP-sensitive potassium channels $(K_{ATP}\;channels)$ is yet to fully explained. This study was carried out to investigate the effects of intracellular application of monocarboxylates (acetate, formate, lactate, and pyruvate) on $K_{ATP}$ channels in isolated rabbit ventricular myocytes. Single channel currents of $K_{ATP}$ channels were recorded using the excised inside-out or permeabilized attached (open-cell) patch-clamp technique at room temperature. Intracellular application of acetate, formate and pyruvate led to an inhibition of channel activity, whereas intracellular application of lactate increased channel activity. These effects were reversible upon washout. Analysis of single channel kinetics showed that monocarboxylates did not affect open-time constant and close-time constant. These results suggest that monocarboxylates participate in modulating $K_{ATP}$ channels activity in cardiac cells and that modulation of $K_{ATP}$ channels activity may resolve the discrepancy between the low $K_i$ in excised membrane patches and high levels of intracellular ATP concentration during myocardial ischemia or hypoxia.

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Inhibition of Pacemaker Activity of Interstitial Cells of Cajal by Hydrogen Peroxide via Activating ATP-sensitive $K^+$ Channels

  • Choi Seok;Parajuli Shankar Prasad;Cheong Hyeon-Sook;Paudyal Dilli Parasad;Yeum Cheol-Ho;Yoon Pyung-Jin;Jun Jae-Yeoul
    • The Korean Journal of Physiology and Pharmacology
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    • 제11권1호
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    • pp.15-20
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    • 2007
  • To investigate whether hydrogen peroxide($H_2O_2$) affects intestinal motility, pacemaker currents and membrane potential were recorded in cultured interstitial cells of Cajal(ICC) from murine small intestine by using a whole-cell patch clamp. In whole cell patch technique at $30^{\circ}C$, ICC generated spontaneous pacemaker potential under current clamp mode(I=0) and inward currents(pacemaker currents) under voltage clamp mode at a holding potential of -70 mV. When ICC were treated with $H_2O_2$ in ICC, $H_2O_2$ hyperpolarized the membrane potential under currents clamp mode and decreased both the frequency and amplitude of pacemaker currents and increased the resting currents in outward direction under voltage clamp mode. Also, $H_2O_2$ inhibited the pacemaker currents in a dose-dependent manner. Because the properties of $H_2O_2$ action on pacemaker currents were same as the effects of pinacidil(ATP-sensitive $K^+$ channels opener), we tested the effects of glibenclamide(ATP-sensitive $K^+$ channels blocker) on $H_2O_2$ action in ICC, and found that the effects of $H_2O_2$ on pacemaker currents were blocked by co- or pre- treatment of glibenclamide. These results suggest that $H_2O_2$ inhibits pacemaker currents of ICC by activating ATP-sensitive $K^+$ channels.

세포외 분비시 막 캐패시턴스를 측정하기 위한 위상감지법(phase detector technique)의 이론적 분석. (Theoretical Analysis of Phase Detector Technique for the Measurement of Cell Membrane Capacitance During Exocytosis)

  • Cha, Eun-Jong;Goo, Yong-Sook;Lee, Tae-Soo
    • 한국의학물리학회지:의학물리
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    • 제3권2호
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    • pp.43-57
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    • 1992
  • 위상감지법(phase detector technique)은 세포의 막 캐패시턴스(membrane capacitance)를 실시간적으로 측정할 수 있는 유일한 방법이나 측정이 행해지는 동안 세포의 상태가 끊임없이 변화하기 때문에 피할 수 없는 측정오차가 존재한다. 본 연구는 이 오차의 근원을 분석하여 위상감지법의 실용한계를 규정하고자 하였다. 이론적 분석에 기초하여 다음과 같은 사실을 밝힐 수 있었다. 1) access conductance와 membrane conductance의 변화에 기인하는 측정오차를 줄이기 위해서는 초기 위상치를 올바로 선택하여야 한다. 2) 이 때 세포를 여기시키기 위해 인가하는 전압의 주파수를 알맞게 선택하여야 한다. 3) 그러나 초기 위상치가 정해진 이후의 위상 변화는 막 캐패시턴스의 측정에 큰 영향을 미치지 않는다. 4) 초기 위상을 적절히 선택하였다 하더라도 세포외 분비시 막 캐패시턴스가 크게 증가하는 경우에는 비례상수에 오차가 발생한다. 이 때 발생하는 오차는 측정기간 동안 비례상수를 되풀이하여(iteration) 보정함으로써 방지할 수 있다. 이상의 결과는 향후 위상감지법을 사용할 때 유용한 설용한계를 제공하리라 생각된다.

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