• 제목/요약/키워드: patch-clamp technique

검색결과 155건 처리시간 0.03초

Altered Delayed Rectifier $K^+$ Current of Rabbit Coronary Arterial Myocytes in Isoproterenol-Induced Hypertrophy

  • Kim, Na-Ri;Han, Jin;Kim, Eui-Yong
    • The Korean Journal of Physiology and Pharmacology
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    • 제5권1호
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    • pp.33-40
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    • 2001
  • The aim of present study was to define the cellular mechanisms underlying changes in delayed rectifier $K^+\;(K_{DR})$ channel function in isoproterenol-induced hypertrophy. It has been proposed that $K_{DR}$ channels play a role in regulation of vascular tone by limiting membrane depolarization in arterial smooth muscle cells. The alterations of the properties of coronary $K_{DR}$ channels have not been studied as a possible mechanism for impaired coronary reserve in cardiac hypertrophy. The present study was carried out to compare the properties of coronary $K_{DR}$ 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 study are as follows: (1) the $K_{DR}$ current density was decreased without changes of the channel kinetics in isoproterenol-induced hypertrophy; (2) the sensitivity of coronary $K_{DR}$ channels to 4-AP was increased in isoproterenol-induced hypertrophy. From the above results, we suggest for the first time that the alteration of $K_{DR}$ channels may limit vasodilating responses to several stimuli and may be involved in impaired coronary reserve in isoproterenol-induced hypertrophy.

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Effect of Capsaicin on Delayed Rectifier $K^+$ Current in Adult Rat Dorsal Root Ganglion Neurons

  • Hahn, Jung-Hyun;Chung, Sung-Kwon;Bang, Hyo-Weon
    • The Korean Journal of Physiology and Pharmacology
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    • 제4권1호
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    • pp.9-14
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    • 2000
  • $K^+$ currents play multiple roles in the excitability of dorsal root ganglion (DRG) neurons. Influences on these currents change the shape of the action potential, its firing threshold and the resting membrane potential. In this study, whole cell configuration of patch clamp technique had been applied to record the blocking effect of capsaicin, a lipophilic alkaloid, on the delayed rectifier $K^+$ current in cultured small diameter DRG neurons of adult rat. Capsaicin reduced the amplitude of $K^+$ current in dose dependent manner, and the concentration-dependence curve was well described by the Hill equation with $K_D$ value of $19.1{\mu}M.$ The blocking effect of capsaicin was reversible. Capsaicin $(10 {\mu}M)$ shifted the steady-state inactivation curve in the hyperpolarizing direction by about 15 mV and increased the rate of inactivation. The voltage dependence of activation was not affected by capsaicin. These multiple effects of capsaicin may suggest that capsaicin bind to the region of $K^+$ channel, participating in inactivation process.

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Effects of Adenosine on the Ionic Channel Activated by Metabolic Inhibition in Rabbit Ventricular Myocytes

  • Han, Jin;Kim, Eui-Yong;Ho, Won-Kyung;Earm, Yung-E
    • The Korean Journal of Physiology
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    • 제30권1호
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    • pp.1-9
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    • 1996
  • The objective of the present study was to characterize the role of adenosine in regulation of ATP-sensitive $K^+\;channel\;(K_{ATP}\;channel)$ activity in isolated rabbit ventricular myocytes using the patch clamp technique. Internal adenosine had little effects on KaTr channel activity. In an outside-out patch with intrapipette GTP and ATP, external adenosine stimulated $K_{ATP}\;channel$ activity. In an inside-out Patch with intrapipette adenosine, ATP reduced $K_{ATP}\;channel$ activity, and GTP stimulated $K_{ATP}\;channel$ activity. Adenosine receptor activation shifted the half-maximal inhibition Of $K_{ATP}\;channel\;from\;70\;to\;241\;{\mu}m$. These results Suggest that activation of adenosine receptors stimulates $K_{ATP}\;channels$ in rabbit ventricular myocytes by reducing the apparent affinity of the channel for ATP. The effect may be important for activating $K_{ATP}\;channels$ during early phase of myocardial ischemia.

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Inactivation of N-Type Calcium Current in Rat Sympathetic Neurons

  • Lee, Mi-Sun;Goo, Yong-Sook
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2001년도 학술 발표회 진행표 및 논문초록
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    • pp.26-26
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    • 2001
  • Inactivation of N-type calcium current has been reported to be both voltage dependent and Ca$\^$2+/ dependent. We have investigated the effects of Ba$\^$2+/ and Ca$\^$2+/ on N-channel inactivation in rat superior cervical ganglion neurons using the whole cell configuration of patch clamp technique. Inactivation was larger in Ca$\^$2+/ than in Ba$\^$2+/ even with 20 mM BAPTA.(omitted)

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Changes in Intracellular $Ca^{2+}$ Concentration Induced by L-Type $Ca^{2+}$ Channel Current in Guinea-Pig Gastric Myocytes

  • Kim, Ki-Whan
    • 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)

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Protein Kinase Modulates the $GABA_c$ Currents in Cone-horizontal Cell Axon-terminals Isolated from Catfish Retina

  • Paik, Sun-Sook;Lee, Sung-Jong;Jung, Chang-Sub;Bai, Sun-Ho
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 1999년도 학술발표회 진행표 및 논문초록
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    • pp.54-54
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    • 1999
  • Protein kinase modulation of gamma-aminobutyric acid C (GABA$_{c}$) currents in freshly dissociated catfish retinal cone-horizontal cell axon-terminals was studied under voltage clamp with the use of the whole cell patch-clamp technique. Responses to pulses of GABA were monitored in intracellular application of adenosin 3',5'-cycle monophophate (cAMP)-dependent protein kinase (PKA) and protein kinase C (PKC) activators, and their inhibitors or inactive analogues.(omitted)d)

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Effect of Propofol on Ion Channels in Acutely Dissociated Dorsal Raphe Neuron of Sprague-Dawley Rats

  • Lee, Bong-Jae;Kwon, Moo-ll;Shin, Min-Chul;Kim, Youn-Jung;Kim, Chang-Ju;Kim, Soon-Ae;Kim, Ee-Hwa;Chung, Joo-Ho
    • The Korean Journal of Physiology and Pharmacology
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    • 제5권2호
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    • pp.189-197
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    • 2001
  • To investigate propofol's effects on ionic currents induced by ${\gamma}-aminobutyric$ acid (GABA) and glycine as well as on those produced by the nicotinic acetylcholine- and glutamate-responsive channels, rat dorsal raphe neurons were acutely dissociated and the nystatin-perforated patch-clamp technique under voltage-clamp conditions was used to observe their responses to the administration of propofol. Propofol evoked ion currents in a dose-dependent manner, and propofol $(10^{-4}\;M)$ was used to elicit ion currents through the activation of $GABA_A,$ glycine, nicotinic acetylcholine and glutamate receptors. Propofol at a clinically relevant concentration $(10^{-5}\;M)$ potentiated $GABA_A-,$ glycine- and NMDA receptor-mediated currents. The potentiating action of propofol on $GABA_A-,$ glycine- and NMDA receptor-mediated responses involved neither opioid receptors nor G-proteins. Apparently, propofol modulates inhibitory and excitatory neurotransmitter-activated ion channels either by acting directly on the receptors or by potentiating the effects of the neurotransmitters, and this modulation appears to be responsible for the majority of the anaesthetic and/or adverse effects.

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Modulation of the aqueous extract of Bupleuri radix on glycine-induced current in the acutely dissociated rat periaqueductal gray neurons

  • Sung, Yun-Hee;Shin, Mal-Soon;Kim, Tae-Soo;Lee, Sang-Won;Kim, Youn-Jung;Shin, Hye-Sook;Kim, Hong;Kim, Chang-Ju
    • Advances in Traditional Medicine
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    • 제7권5호
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    • pp.549-555
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    • 2008
  • Bupleuri radix (Umbelliferae), the dried root of Bupleurum Chinense DC, has been clinically used to mitigate pain sensation. The descending pain control system consists of three major components, and modulation of pain in the periaqueductal gray is the most extensively studied descending pain control system. However, the relation of Bupleuri radix on the descending pain control system has not been clarified. In the present study, modulation of the aqueous extract of Bupleuri radix on glycine-induced ion current in the acutely dissociated periaqueductal gray neurons was investigated by using nystatin-perforated patch-clamp technique under voltage-clamp condition. In the present results, the glycine-induced ion current was significantly suppressed by 0.1 mg/ml Bupleuri radix, while treatment with $10^{-5}\;M$ naltrexone, opioid receptor antagonist, alleviated Bupleuri radix-induced inhibition on glycine-induced ion current. The present study showed that the aqueous extract of Bupleuri radix may activate descending pain control system through inhibition on glycine-induced ion current in the periaqueductal gray neurons and this effect is mediated by opioid receptors.

Bile Acid Modulation of Gastroinstinal Smooth Muscle Contraction and Ionic Currents

  • Lee, Hye-Kyung;Lee, Kyoung-Hwa
    • The Korean Journal of Physiology and Pharmacology
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    • 제4권4호
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    • pp.333-338
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    • 2000
  • We have examined whether bile acids can affect the electrical and mechanical activities of circular smooth muscle of canine colon and ileum, using isometric tension measurement or patch clamp technique. It was found that a dilution of canine bile $(0.03{\sim}2%\;by\;volume)$ enhanced or inhibited the amplitude of spontaneous contractions. An individual component of bile, deoxycholic acid (DCA) enhanced the frequency and amplitude of the spontaneous contractile activity at $10^{-6}\;M,$ while DCA at $10^{-4}\;M$ inhibited the contraction. Similarly, the response to cholic acid was excitatory at $10^{-5}\;M$ and inhibitory at $3{\times}10^{-4}\;M.$ Taurocholic acid at $10^{-4}\;M$ enhanced the amplitude of muscle contraction. Electrically, canine bile at 1% reversibly depolarized the colonic myocytes under current clamp mode. Bile acids also elicited non-selective cation currents under voltage clamp studies, where $K^+$ currents were blocked and the $Cl^-$ gradient was adjusted so that $E_{Cl}^-$ was equal to -70 mV, a holding potential. The non-selective cation current might explain the depolarization caused by bile acids in intact muscles. Furthermore, the bile acid regulation of electrical and mechanical activities of intestinal smooth muscle may explain some of the pathophysiological conditions accompanying defects in bile reabsorption.

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External pH Effects on Delayed Rectifier $K^+$ Currents of Small Dorsal Root Ganglion Neuron of Rat

  • Kim, Young-Ho;Hahn, Jung-Hyun;Lim, In-Ja;Chung, Sung-Kwon;Bang, Hyo-Weon
    • The Korean Journal of Physiology and Pharmacology
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    • 제2권2호
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    • pp.165-172
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    • 1998
  • Under certain pathophysiological conditions, such as inflammation and ischemia, the concentration of H^+$ ion in the tissue surrounding neurons is changed. Variations in H^+$ concentration are known to alter the conduction and/of the gating properties of several types of ion channels. Several types of K^+$ channels are modulated by pH. In this study, the whole cell configuration of the patch clamp technique has been applied to the recording of the responses of change of external pH on the delayed rectifier K^+$ current of cultured DRG neurons of rat. Outward K^+$ currents were examined in DRG cells, and the Charybdotoxin and Mn^{2+}$ could eliminate Ca^{2+}-dependent$ K^+$ currents from outward K^+$ currents. This outward K^+$ current was activated around -60 mV by step depolarizing pulses from holding potential -70 mV. Outward K^+$ currents were decreased by low external pH. Activation and steady-state inactivation curve were shifted to the right by acidification, while there was small change by alkalization. These results suggest that H^+$ could be alter the sensory modality by changing and modifying voltage-dependent K^+$ currents, which participated in repolarization.

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