• Title/Summary/Keyword: Activation Channels

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Identification of phospholipase Cβ downstream effect on transient receptor potential canonical 1/4, transient receptor potential canonical 1/5 channels

  • Ko, Juyeon;Myeong, Jongyun;Kwak, Misun;Jeon, Ju-Hong;So, Insuk
    • The Korean Journal of Physiology and Pharmacology
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    • v.23 no.5
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    • pp.357-366
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    • 2019
  • $G{\alpha}_q$-coupled receptor stimulation was implied in the activation process of transient receptor potential canonical (TRPC)1/4 and TRPC1/5 heterotetrameric channels. The inactivation occurs due to phosphatidylinositol 4,5-biphosphate ($PI(4,5)P_2$) depletion. When $PI(4,5)P_2$ depletion was induced by muscarinic stimulation or inositol polyphosphate 5-phosphatase (Inp54p), however, the inactivation by muscarinic stimulation was greater compared to that by Inp54p. The aim of this study was to investigate the complete inactivation mechanism of the heteromeric channels upon $G{\alpha}_q$-phospholipase $C{\beta}$ ($G{\alpha}_q-PLC{\beta}$) activation. We evaluated the activity of heteromeric channels with electrophysiological recording in HEK293 cells expressing TRPC channels. TRPC1/4 and TRPC1/5 heteromers undergo further inhibition in $PLC{\beta}$ activation and calcium/protein kinase C (PKC) signaling. Nevertheless, the key factors differ. For TRPC1/4, the inactivation process was facilitated by $Ca^{2+}$ release from the endoplasmic reticulum, and for TRPC1/5, activation of PKC was concerned mostly. We conclude that the subsequent increase in cytoplasmic $Ca^{2+}$ due to $Ca^{2+}$ release from the endoplasmic reticulum and activation of PKC resulted in a second phase of channel inhibition following $PI(4,5)P_2$ depletion.

The Design of 128 Channels Cardiac-Activation Pre-Amplifier (128 채널 심장전기도 전치 증폭기의 설계)

  • Yoo, Sun-Kook;Chang, Byung-Chul;Jung, Dong-Il;Han, Young-Oh
    • The Transactions of the Korean Institute of Electrical Engineers D
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    • v.50 no.11
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    • pp.550-556
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    • 2001
  • The computerized cardiac analysis system, which acquires and analyzes the electrical activation signal propagating along the surface of the heart, is indispensible equipment for the open heart surgery and electrical cardiac study. In this paper, the design requirement and the electrical circuit analysis are performed to construct the multi-channel cardiac activation pre-amplifier necessary for a signal conditioning circuit. The general 64 channel configuration is expanded into 128 channels to enhance the spatial resolution on the mapped surface of the heart. The 128 channels pre-amplifier consists of input circuit, differential amplifier, right leg driven circuit and isolation part. It has distinct features; high voltage protection, leakage current limitation, isolation and the maximization of common mode rejection ratio with respect to the half-cell potential difference due to different electrode materials. The final pre-amplifier circuit is assembled with 8 boards, each of which composing of 16 channels.

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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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    • v.4 no.4
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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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Cytosolic domain regulates the calcium sensitivity and surface expression of BEST1 channels in the HEK293 cells

  • Kwon Woo Kim;Junmo Hwang;Dong-Hyun Kim;Hyungju Park;Hyun-Ho Lim
    • BMB Reports
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    • v.56 no.3
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    • pp.172-177
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    • 2023
  • BEST family is a class of Ca2+-activated Cl- channels evolutionary well conserved from bacteria to human. The human BEST paralogs (BEST1-BEST4) share significant amino acid sequence homology in the N-terminal region, which forms the transmembrane helicases and contains the direct calcium-binding site, Ca2+-clasp. But the cytosolic C-terminal region is less conserved in the paralogs. Interestingly, this domain-specific sequence conservation is also found in the BEST1 orthologs. However, the functional role of the C-terminal region in the BEST channels is still poorly understood. Thus, we aimed to understand the functional role of the C-terminal region in the human and mouse BEST1 channels by using electrophysiological recordings. We found that the calcium-dependent activation of BEST1 channels can be modulated by the C-terminal region. The C-terminal deletion hBEST1 reduced the Ca2+-dependent current activation and the hBEST1-mBEST1 chimera showed a significantly reduced calcium sensitivity to hBEST1 in the HEK293 cells. And the C-terminal domain could regulate cellular expression and plasma membrane targeting of BEST1 channels. Our results can provide a basis for understanding the C-terminal roles in the structure-function of BEST family proteins.

Expression of Low Voltage-Activated $Ca^{2+}$ Channels in Xenopus Oocytes

  • Lee, Jung-Ha;Han, Dong-Pyo
    • Journal of Microbiology and Biotechnology
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    • v.11 no.4
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    • pp.614-618
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    • 2001
  • Low-threshold T-type $Ca^{2+}$ channels are distinctive voltage-operated gates for external $Ca^{2+}$ entry around a resting membrane potential due to their low voltage activation. These phenomena have already been extensively studied due to their relevance in diverse physiological functions. Recently, three T-type $Ca^{2+}$ channel ${\alpha}$$_1$subunits were cloned and their biophysical properties were characterized after expression in mammalian expression systems. In this study, ${\alpha_IG} and {\alpha_IH}$ low-threshold $Ca^{2+}$ channels were expressed and characterized in Xenopus oocytes after adding 5' and 3'untranslated portions of a Xenopus ${\beta}$ globin to improve their expression levels. The added portions dramatically enhanced the expression levels of the ${\alpha_IG} and {\alpha_IH}$ T-type channels. When currents were recorded in 10 mM $Ba^{2+}$ as the charge carrier, the activation thresholds were about -60 mV, peak currents appeared at -20 mV, and the reversal potentials were between +40 and +45. The activation time constants were very similar to each other, while the inactivation time constants of the ${\alpha_IG}$ currents were smaller than those of ${\alpha_IH}$. Taken together, the electrophysiological properties of the ${\alpha_IG} and {\alpha_IH}$ channels expressed in Xenopus oocytes were similar to the previously reported characteristics of low-threshold $Ca^{2+}$ channel currents.

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Transient receptor potential melastatin type 7 channels are involved in zinc-induced apoptosis in gastric cancer

  • Kim, Byung-Joo
    • Animal cells and systems
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    • v.15 no.2
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    • pp.123-130
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    • 2011
  • Transient receptor potential melastatin 7 (TRPM7) channels are novel $Ca^{2+}$-permeable non-selective cation channels that are ubiquitously expressed. Activation of TRPM7 channels has been shown to be involved in the survival of gastric cancer cells. Here we show evidence suggesting that TRPM7 channels play an important role in $Zn^{2+}$- mediated cellular injury. Using a combination of electrophysiology, pharmacological analysis, small interfering RNA (siRNA) methods and cell death assays, we showed that activation of TRPM7 channels augmented $Zn^{2+}$-induced apoptosis of AGS cells, the most common human gastric adenocarcinoma cell line. The $Zn^{2+}$-mediated cytotoxicity was inhibited by the non-specific TRPM7 blockers $Gd^{3+}$ or 2 aminoethoxydiphenyl borate (2-APB) and TRPM7 specific siRNA. In addition, we showed that overexpression of TRPM7 channels in HEK293 cells increased $Zn^{2+}$- induced cell injury. Thus, TRPM7 channels may represent a novel target for physiological disorders where $Zn^{2+}$ toxicity plays an important role.

The imaging of 128 channels cardiac activation signal (128채널 심장전기도 신호의 영상화)

  • Han, Young-Oh
    • The Journal of the Korea institute of electronic communication sciences
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    • v.5 no.2
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    • pp.133-137
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    • 2010
  • In this study, The imaging of 128 channels cardiac activation signal is implemented. It is performed within one minutes and very useful in open-heart operation with analysis in a short time. Especially the keeping of data and the moving of system is facility because it was implemented with being based on PC. The cardiologist can call data and analysis them anytime after a surgery.

The Downregulation of Somatic A-Type $K^+$ Channels Requires the Activation of Synaptic NMDA Receptors in Young Hippocampal Neurons of Rats

  • Kang, Moon-Seok;Yang, Yoon-Sil;Kim, Seon-Hee;Park, Joo-Min;Eun, Su-Yong;Jung, Sung-Cherl
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.2
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    • pp.135-141
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    • 2014
  • The downregulation of A-type $K^+$ channels ($I_A$ channels) accompanying enhanced somatic excitability can mediate epileptogenic conditions in mammalian central nervous system. As $I_A$ channels are dominantly targeted by dendritic and postsynaptic processings during synaptic plasticity, it is presumable that they may act as cellular linkers between synaptic responses and somatic processings under various excitable conditions. In the present study, we electrophysiologically tested if the downregulation of somatic $I_A$ channels was sensitive to synaptic activities in young hippocampal neurons. In primarily cultured hippocampal neurons (DIV 6~9), the peak of $I_A$ recorded by a whole-cell patch was significantly reduced by high KCl or exogenous glutamate treatment to enhance synaptic activities. However, the pretreatment of MK801 to block synaptic NMDA receptors abolished the glutamate-induced reduction of the $I_A$ peak, indicating the necessity of synaptic activation for the reduction of somatic $I_A$. This was again confirmed by glycine treatment, showing a significant reduction of the somatic $I_A$ peak. Additionally, the gating property of $I_A$ channels was also sensitive to the activation of synaptic NMDA receptors, showing the hyperpolarizing shift in inactivation kinetics. These results suggest that synaptic LTP possibly potentiates somatic excitability via downregulating $I_A$ channels in expression and gating kinetics. The consequential changes of somatic excitability following the activity-dependent modulation of synaptic responses may be a series of processings for neuronal functions to determine outputs in memory mechanisms or pathogenic conditions.

Activation of ATP-sensitive Potassium Channels by the Predominant Metabolite of Isoflurane in Rabbit Ventricular Myocytes

  • Han, Jin;Kim, Na-Ri;Kim, Eui-Yong;Kim, Sung-Ju;Cho, Kang-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.2
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    • pp.165-175
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    • 2001
  • Background: Recent in vivo experimental evidence suggests that isoflurane-induced cardioprotection may involve $K_{ATP}$ channel activation. However, it was demonstrated that isoflurane inhibited $K_{ATP}$ channel activities in the inside-out patch mode. To explain this discrepancy, the present investigation tested the hypothesis that a metabolite of isoflurane, trifluoroacetic acid (TFA), contributes to isoflurnae-induced cardioprotection via $K_{ATP}$ channel activation during myocardial ischemia and reperfusion. Methods: Single ventricular myocytes were isolated from rabbit hearts by an enzymatic dissociation procedure. Patch-clamp techniques were used to record single-channel currents. $K_{ATP}$ channel activities were assessed before and after the application of TFA with the inside-out patch mode. Results: TFA enhanced channel activity in a concentration-dependent fashion. The concentration of TFA for half-maximal activation and the Hill coefficient were 0.03 mM and 1.2, respectively. TFA did not affect the single channel conductance of $K_{ATP}$ channels. Analysis of open and closed time distributions showed that TFA increased burst duration and decreased the interburst interval without changes in open and closed time distributions shorter than 5 ms. TFA diminished ATP sensitivity of $K_{ATP}$ channels in a concentration-response relationship for ATP. Conclusions: TFA, a metabolite of isoflurane, enhanced $K_{ATP}$ channel activity in a concentration-dependent fashion. These results imply that TFA could mediate isoflurane-induced cardioprotection via $K_{ATP}$ channel activation during myocardial ischemia and reperfusion.

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The design of filters for multi-channel cardiac activation main amplifier (다중 채널 심장전기도 주 증폭기를 위한 필터 설계)

  • Han, Young-Oh
    • The Journal of the Korea institute of electronic communication sciences
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    • v.5 no.2
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    • pp.180-184
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    • 2010
  • In this paper, the filter design and simulation are performed to construct the multi-channel cardiac activation main amplifier. The main amplifier consists of sample and holder, variable high pass filter, notch filter, variable low pass filter. The general 64 channel configuration is expanded into 128 channels to enhance the spatial resolution and the filter is designed for 128 channels cardiac activation main amplifier.