• Title/Summary/Keyword: Patch-clamp

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Decursin from Angelica gigas Nakai Blocks hKv1.5 Channel

  • Kwak, Yong-Geun;Choi, Bok-Hee;Kim, Dae-Keun;Eun, Jae-Soon
    • Biomolecules & Therapeutics
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    • v.19 no.1
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    • pp.33-37
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    • 2011
  • Decursin was purified from Angelica gigas Nakai, and its effects on the human Kv1.5 (hKv1.5) currents were recorded in mouse fibroblasts ($Ltk^-$ cells) by whole-cell patch-clamp technique. Decursin inhibited hKv1.5 current in a concentration-dependent manner, with an $IC_{50}$ value of $2.7\;{\mu}M$ at +60 mV. Decursin accelerated the inactivation kinetics of the hKv1.5 channel, and slowed the deactivation kinetics of the hKv1.5 current, resulting in a tail crossover phenomenon. Also, decursin inhibited the hKv1.5 current in a use-dependent manner. These results strongly suggest that decursin is a kind of open-channel blocker of the hKv1.5 channel.

GATING MECHANISM AND VOLTAGE-DEPENDENT BLOCK BY EXTERNAL DIVALENT CATIONS OF THE DELAYED RECTIFIER K CHANNEL IN RABBIT SINO-ATRIAL NODE CELLS

  • Ho, Won-Kyung;Lee, Suk-Ho;Earm, Yung-E
    • Proceedings of the Korean Biophysical Society Conference
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    • 1996.07a
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    • pp.5-5
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    • 1996
  • In sino-atrial node cells which act as the normal pacemaker of the heart, K conductance in resting state is minimal due to the absence of inward rectifier K channels K conductance only increases when the membrane is depolarized by the activation of the delayed rectifier K current I$\_$k/. In the present study, we investigated the gating mechanism of$\_$k/ using the whole cell patch clamp technique in isolated single sinoatrial cells of the rabbit. (omitted)

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CONTRIBUTION OF DIFFERENT TYPES OF $Ca^{2+}$ CHANNELS TO CATECHOLAMINE SECRETION IN RAT CHROMAFFIN CELLS

  • Goo, Yong-Sook;Jina Roh;Lee, Jung-Hwa;Cha, Eun-Jong
    • Proceedings of the Korean Biophysical Society Conference
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    • 1996.07a
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    • pp.42-42
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    • 1996
  • Adrenal chromaffin cells secrete catecholamine in response to acetylcholine. The secretory response has absolute requirement for extracellular calcium, indicating that $Ca^{2+}$ influx through voltage operated $Ca^{2+}$ channels (VOCC) is the primary trigger of the secretion cascade. Although the existence of various types of $Ca^{2+}$ channels has been explore using patch clamp techique in adrenal chromaffin cells, the contribution of different types of $Ca^{2+}$ channels to catecholamine secretion remains to be establised. (omitted)omitted)

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Purinoceptor and Intracellular $Ca^{2+}$ Regulation in Rat Prostate N euronencocrine Cells

  • Kim, Jun-Hee;Nam, Joo-Hyun;Uhm, Dae-Yong;Kim, Sung-Joon
    • Proceedings of the Korean Biophysical Society Conference
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    • 2002.06b
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    • pp.45-45
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    • 2002
  • Extracellular ATP regulates a wide range of cellular function including the growth of prostate gland. Purinoceptors (ATP receptors) are divided into P2X (ligand-gated ion channels) and P2Y (G-protein-coupled receptor) subfamilies. In the present study, we investigated the types of purinoceptors in rat prostate neuroendocrine (RPNE) cells using whole-cell patch clamp technique, intracellular $Ca^{2+}$ measurement and RT-PCR analysis.(omitted)d)

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Actin Filaments Regulate the Stretch Sensitivity of Large Conductance $Ca^{2+}$-Activated $K^+$ Channel in Rabbit Coronary Arterial Smooth Muscle Cells

  • Lin Piao;Earm, Yung-E;Wonkyung Ho
    • Proceedings of the Korean Biophysical Society Conference
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    • 2002.06b
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    • pp.35-35
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    • 2002
  • The large conductance $Ca^{2+}$ -activated $K^{+}$ channels ($BK_{Ca}$) in vascular smooth muscle have been considered to function as a negative feedback in pressure-induced vasoconstriction. In the present study, the function of cytoskeletons in the regulation of $BK_{Ca}$ and its stretch sensitivity was investigated. Using the inside-out patch clamp technique, we recorded single channel activities of $BK_{Ca}$ with 150 mM KCl in the bath solution (pCa=6.5).(omitted)itted)

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Lithospermic acid modulate the Action potential duration by increasing Ica current in the rat ventricular myocyte

  • An, Seong-Hun;Kang, Dae-Gill;Lee, Ho-Sup;Lee, Suk-Ho;Earm, Yung-E
    • Proceedings of the Korean Biophysical Society Conference
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    • 2001.06a
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    • pp.55-55
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    • 2001
  • We observed the APD of rat ventricle myocyte and the effects of Lithospermic acid that was separated at Salvia miltiorrhiza having used in Oriental medicine by using classical whole cell patch clamp technique. We classified APD into APD30mV, APD0mV, APD-50mV, APD-60mV by cell membrane potential and the mean of cell resting membrane potential was -69.44${\pm}$1.72 mV.(omitted)

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Inhibition of Acetylcholine-activated $K^+$ Current by Chelerythrine and Bisindolylmaleimide I in Atrial Myocytes from the Mice

  • Hana Cho;Youm, Jae-Boum;Earm, Yung-E;Ho, Won-Kyung
    • Proceedings of the Korean Biophysical Society Conference
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    • 2001.06a
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    • pp.54-54
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    • 2001
  • The effects of protein kinase C inhibitors, chelerythrine and bisindolylmaleimide I, on acetylcholine activated $K^{+}$ currents ( $I_{KACh}$) were examined in atrial myocytes of mice using patch clamp technique. Chelerythrine and bisindolylmaleimide I inhibited $I_{KACh}$ in reversible and dose-dependent manners. Half maximal effective concentrations were 0.49 $\pm$ 0.01 $\mu$M for chelerythrine and 98.69 $\pm$ 12.68 nM for bisindolylmaleimide I.(omitted)

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Role of Protein Kinase C on Norepinephrine Induced Inhibition of Calcium Current in Rat Sympathetic Neurons (흰쥐 교감신경세포에서 Norepinephrine 에 의한 칼슘전류 억제에 미치는 Protein Kinase C 의 역할)

  • ;Keith S. Elmslie
    • Progress in Medical Physics
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    • v.11 no.1
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    • pp.29-38
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    • 2000
  • The signal transduction pathway for most neurotransmitter induced inhibition of $Ca^{2+}$ channels in sympathetic neurons involves a G-protein mediated, membrane-delimited mechanism without the participation of any known protein kinase. However, activation of protein kinase C (PKC) has been proposed as one of the intracellular mechanisms mediating some neurotransmitter induced $Ca^{2+}$ channel inhibition. In the present study, we investigated the effects of phorbol-12, 13-dibutyrate (PDBu) on $Ca^{2+}$ channel currents of acutely dispersed neurons from adult rat superior cervical ganglion (SCG) neurons using whole cell variant of the patch clamp technique. PDBu (500 nM), the activator of PKC, increased $Ca^{2+}$ channel currents and retarded the deactivation of tail currents. The effects of PDBu were voltage dependent and the maximal increase in the current amplitudes was observed between -10 to 10 mV (n=4). PDBu attenuated $Ca^{2+}$ current inhibition induced by norepinephrine (NE), which modulates $Ca^{2+}$ channels via a pertussis toxin (PTX)-sensitive pathway. Inhibition of PDBu by staurosporine (1 $\mu$M) blocked the effects of PDBu on current amplitudes and NE-induced G-protein mediated inhibition of $Ca^{2+}$ currents. Further experiment should be done to know if G-protein or $Ca^{2+}$ channel itself is the target of PKC phosphorvlation.phosphorvlation.

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Characterization of Intermediate Conductance $K^+$ Channels in Submandibular Gland Acinar Cells

  • Cho, Sung-Man;Piao, Zheng-Gen;Kim, Yoon-Bae;Kim, Joong-Soo;Park, Kyung-Pyo
    • The Korean Journal of Physiology and Pharmacology
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    • v.6 no.6
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    • pp.305-309
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    • 2002
  • There are some evidences that $K^+$ efflux evoked by muscarinic stimulation is not mainly mediated by large conductance $K^+$ (BK) channels in salivary gland. In this experiment, we therefore characterised non BK channels in rat submandibular gland acinar cells and examined the possibility of agonist effect on this channel using a patch clamp technique. Two types of $K^+$ channels were observed in these cells. BK channels were observed in 3 cells from total 6 cells and its average conductance was $152{\pm}7$ pS (n=3). The conductance of the another types of $K^+$ channel was estimated as $71{\pm}7$ pS (n=6). On the basis of the conductance of this channel, we defined this channel as intermediate conductance $K^+$ (IK) channels, which were observed from all 6 cells we studied. When we increased $Ca^{2+}$ concentration of the bath solution in inside-out mode, the IK channel activity was greatly increased, suggesting this channel is $Ca^{2+}$ sensitive. We next examined the effect of carbachol (CCh) and isoproterenol on the activity of the IK channels. $10^{-5}$ M isoproterenol significantly increased the open probability (Po) from $0.08{\pm}0.02$ to $0.21{\pm}0.03$ (n=4, P<0.05). Application of $10^{-5}$ M CCh also increased Po from $0.048{\pm}0.03$ to $0.55{\pm}0.33$ (n=5, P<0.05) at the maximum channel activity. The degree of BK channel activation induced by the same concentration of CCh was lower than that of IK channels; Po value was $0.011{\pm}0.003$ and $0.027{\pm}0.005$ in control and during CCh stimulation (n=3), respectively. The result suggests that IK channels exist in salivary acinar cells and its channel activity is regulated by muscaricinic and ${\beta}-adrenergic$ agonist. We conclude that IK channels also play a putative role in secretion as well as the BK channels in rat submandibular gland acinar cells.

Effect of Phorbol ester on $K^+$channel in an G292 osteoblast-like cell (G292 세포에서 $K^+$통로에 대한 phorbol ester의 효과)

  • Kim, Mi-Kyung;Park, Su-Byung
    • The korean journal of orthodontics
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    • v.32 no.3 s.92
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    • pp.227-234
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    • 2002
  • In order to investigate the action mechanism of protein kinase C on $K^+$ channel in osteoblastic cell, effects of phorbol 12, 13-dibutyrate on human osteoblast-like cells (G292) were studied by patch clamp technique with cell-attacked configuration. 111 this experiment, 45pS ion channel was dominant in G292 cell line according to their approximate conductances in symmetrical 140mM KCl saline at holding potential of 60mV. In torrent-voltage relationship, reversal potential was 5.5mV at the condition of potassium enriched saline in the pipette and -27 mV at the condition of standard extracellular saline In the pipette. Phorbol 12, 13-dibutyrate 10nM increased the open probability of 45pS channel and staurosporine, an inhibitor of protein kinase C, suppressed this effect. Phorbol 12,13-dibutyrate moved the reversal potential of 45pS channel to more negative potential and increased the single channel current at the same membrame potential. In order to check the activation of protein kinase C in G292 cell by phorbol 12,13-dibutyrate, western blot of protein kinase C was performed. Phorbol 12,13-dibutyrate $0.1{\mu}M$ translocated protein kinase C from cellular compartment to membrane compartment of the cell. These findings suggest that phorbol 12,13-dibutyrate, one of phorbol esters, activate 45pS channel In G292 cell and affect cell membrane potential, that regulate cellular function.