1.Potassium Currents in Isolated Deiters' Cells of Guinea Pig.
Jong Woo CHUNG ; Eui Chol NAM ; Won Tae KIM ; Jae Boum YOUM ; Chae Hun LEEM
The Korean Journal of Physiology and Pharmacology 2013;17(6):537-546
Deiters' cells are the supporting cells in organ of Corti and are suggested to play an important role in biochemical and mechanical modulation of outer hair cells. We successfully isolated functionally different K+ currents from Deiters' cells of guinea pig using whole cell patch clamp technique. With high K+ pipette solution, depolarizing step pulses activated strongly outward rectifying currents which were dose-dependently blocked by clofilium, a class III anti-arrhythmic K+ channel blocker. The remaining outward current was transient in time course whereas the clofilium-sensitive outward current showed slow inactivation and delayed rectification. Addition of 5 mM tetraethylammonium (TEA) further blocked the remaining current leaving a very fast inactivating transient outward current. Therefore, at least three different types of K+ current were identified in Deiters' cells, such as fast activating and fast inactivating current, fast activating slow inactivating current, and very fast inactivating transient outward current. Physiological role of them needs to be established.
Animals
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Ear, Inner
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Guinea Pigs*
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Guinea*
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Hair
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Hearing
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Organ of Corti
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Pharmacology
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Potassium Channels
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Potassium*
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Quaternary Ammonium Compounds
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Tetraethylammonium
2.Calcium-dependent chloride channels in plasma membrane of oocytes from toad, Bufo bufo gargarizans.
Yan-Li PI ; Ji-Hua MA ; Pei-Hua ZHANG ; Jing-Jing DUAN
Acta Physiologica Sinica 2006;58(5):471-476
In this paper, membrane current properties of the fully-grown oocytes from toad, Bufo bufo gargarizans, were studied by using two-microelectrode voltage clamp technique. Axion of adult female toad was destroyed, and then ovarian lobes containing oocytes in stage I to VI were removed and incubated in Ca(2+)-free ND96 solution with collagenase (1.5 mg/ml) for 1 h. Subsequently, the oocytes were washed in Ca(2+)-free ND96 solution for 10 min to completely remove the follicular layer. For the experiments only the oocytes in stage V and VI were selected and used during 1 to 5 d. The membrane was depolarized from a holding potential of -80 mV to +60 mV in 10 mV step. It was found that a sustained outward current was elicited by depolarization. Potassium channel blockers (tetraethylammonium chloride, TEA, 10 mmol/L and 4-aminopyridine, 4-AP, 10 mmol/L) reduced the outward current to (23.4+/-0.72)% of the maximum. However, further addition of chloride channel blocker (5-nitro-2, 3-phenypropylamino benzoate, NPPB, 30 micromol/L) could almost completely block the outward current to (2.1+/-0.08)% of the maximum. In the presence of TEA and 4-AP, removal of extracellular Ca(2+) or adding verapamil (40 micromol/L), could also reduce the outward current to (2.2+/-0.04) % and (3.1+/-0.15) % of the maximum, respectively. It is concluded that calcium-dependent chloride channels exist in plasma membrane of Bufo bufo gargarizans oocytes, besides potassium channels.
4-Aminopyridine
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toxicity
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Animals
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Bufo bufo
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Calcium
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metabolism
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Cell Membrane
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metabolism
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Chloride Channels
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drug effects
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physiology
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Female
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Nitrobenzoates
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pharmacology
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Oocytes
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metabolism
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Tetraethylammonium Compounds
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pharmacology
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Verapamil
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pharmacology
3.Effects of vanadate on vascular contractility and membrane potential in the rabbit aorta.
Sang Man CHUNG ; Duck Sun AHN ; Hye Sun SEOK ; Yong JEONG ; Bok Soom KANG
Yonsei Medical Journal 1992;33(1):14-23
Isolated rabbit aortic ring with intact endothelial cell preparations precontracted with NE (10(-7) M) were relaxed by vanadate in a dose dependent manner (from 0.2 to 2 mM). Application of vanadate and ACh during the tonic phase of high K+(100 mM)-induced contraction showed a slight relaxation in contrast to that in NE-induced contraction, but sodium nitroprusside (10 microM) more effectively relaxed the aortic ring preparations in high K+ contraction than that of vanadate. Vanadate-induced relaxation in NE-contracted aortic rings was reversed by application of BaCl2 (50 microM) or glibenclamide (10 microM). Furthermore, Vanadate hyperpolarized membrane potential of smooth muscle cells in endothelium-intact aortic strips and this effect was abolished by application of glibenclamide. The above results suggest that vanadate release EDHF (Endothelium-Derived Hyperpolarizing Factor), in addition to EDRF (Endothelium-Derived Relaxing Factor) from endothelial cell. This EDHF hyperpolarize the smooth muscle cell membrane potential via opening of the ATP-sensitive K+ channel and close a voltage dependent Ca++ channel. So it is suggested that the vanadate-induced relaxation of rabbit thoracic aortic rings may be due to the combined effects of EDRF and EDHF.
Animal
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Aorta/drug effects/physiology
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In Vitro
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Membrane Potentials/drug effects
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Potassium/pharmacology
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Potassium Channels/physiology
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Rabbits
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Support, Non-U.S. Gov't
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Tetraethylammonium Compounds/pharmacology
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Vanadates/*pharmacology
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Vasodilation/*drug effects
4.Sodium-activated Potassium Current in Guinea pig Gastric Myocytes.
Young Chul KIM ; Jae Hoon SIM ; Tong Mook KANG ; Hikaru SUZUKI ; Seung Ryul KIM ; Seong Chun KWON ; Wen Xie XU ; Sang Jin LEE ; Ki Whan KIM
Journal of Korean Medical Science 2007;22(1):57-62
This study was designed to identify and characterize Na+ -activated K+ current (I(K(Na))) in guinea pig gastric myocytes under whole-cell patch clamp. After whole-cell configuration was established under 110 mM intracellular Na+ concentration ([Na+]i) at holding potential of -60 mV, a large inward current was produced by external 60 mM K+([K+] degree). This inward current was not affected by removal of external Ca2+. K+ channel blockers had little effects on the current (p>0.05). Only TEA (5 mM) inhibited steady-state current to 68+/-2.7% of the control (p<0.05). In the presence of K+ channel blocker cocktail (mixture of Ba2+, glibenclamide, 4-AP, apamin, quinidine and TEA), a large inward current was activated. However, the amplitude of the steadystate current produced under [K+]degree (140 mM) was significantly smaller when Na+ in pipette solution was replaced with K+ - and Li+ in the presence of K+ channel blocker cocktail than under 110 mM [Na+]i. In the presence of K+ channel blocker cocktail under low Cl- pipette solution, this current was still activated and seemed K+ -selective, since reversal potentials (E(rev)) of various concentrations of [K+]degree-induced current in current/voltage (I/V) relationship were nearly identical to expected values. R-56865 (10-20 microgram), a blocker of IK(Na), completely and reversibly inhibited this current. The characteristics of the current coincide with those of IK(Na) of other cells. Our results indicate the presence of IK(Na) in guinea pig gastric myocytes.
Tetraethylammonium Compounds/pharmacology
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Stomach/*physiology
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Sodium/metabolism/*pharmacology
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Potassium Channels/*physiology
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Potassium Channel Blockers/pharmacology
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Myocytes, Smooth Muscle/*physiology
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Membrane Potentials
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Male
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Guinea Pigs
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Female
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Chlorides/pharmacology
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Calcium/metabolism
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Animals
5.Effect of nitric oxide-induced tyrosine phosphorylation of calcium-activated potassium channel alpha subunit on vascular hyporesponsiveness in rats.
Rong ZHOU ; Liang-ming LIU ; De-yao HU
Chinese Journal of Traumatology 2005;8(4):209-215
OBJECTIVETo study the effect of nitric oxide-induced tyrosine phosphorylation of large-conductance calcium-activated potassium (BK(Ca)) channel alpha subunit on vascular hyporesponsiveness in rats.
METHODSA total of 46 Wistar rats of either sex, weighing 250 g +/- 20 g, were used in this study. Models of vascular hyporesponsiveness induced by hemorrhagic shock (30 mm Hg for 2 hours) in vivo and by L-arginine in vitro were established respectively. The vascular responsiveness of isolated superior mesenteric arteries to norepinephrine was observed. Tyrosine phosphorylation of BK(Ca) alpha subunit was evaluated with methods of immunoprecipitation and Western blotting.
RESULTSIn the smooth muscle cells of the superior mesenteric arteries, the expression of BK(Ca) alpha subunit tyrosine phosphorylation increased following hemorrhagic shock, and L-arginine could induce BK(Ca )channel alpha subunit tyrosine phosphorylation in a time- and dose-dependent manner. L-NAME (Nomega-nitro-L-arginine-methyl-ester), a nitric oxide synthetase inhibitor, could partly restore the decreased vasoresponsiveness of the superior mesenteric arteries after hemorrhagic shock in rats. Down-regulating the protein tyrosine phosphorylation with genistein, a widely-used special protein tyrosine kinase inhibitor, could partly improve the decreased vasoresponsiveness of the superior mesenteric arteries induced by L-arginine in vitro, while up-regulating the protein tyrosine phosphorylation with Na(3)VO(4), a protein tyrosine phosphatase inhibitor, could further decrease the nitric oxide-induced vascular hyporesponsiveness, which could be partly ameliorated by 0.1 mmol/L tetrabutylammonium chloride (TEA), a selective BK(Ca )inhibitor at this concentration.
CONCLUSIONSNitric oxide can induce the tyrosine phosphorylation of BK(Ca) alpha subunit, which influences the vascular hyporesponsiveness in hemorrhagic shock rats or induced by L-arginine in vitro.
Animals ; Arginine ; pharmacology ; Cells, Cultured ; Female ; Male ; Mesenteric Artery, Superior ; drug effects ; physiopathology ; Nitric Oxide ; physiology ; Phosphorylation ; Potassium Channels, Calcium-Activated ; metabolism ; Protein Subunits ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; metabolism ; Tetraethylammonium Compounds ; pharmacology ; Tyrosine ; metabolism ; Vasoconstriction ; drug effects
6.Roles of potassium channel in effects of resveratrol on isolated myocardial contractility and heart rate research in guinea pig.
Gui-ying WANG ; Cui-miao SONG ; Li-nan ZHANG ; Qian LI ; Hua YUE ; Jing-kun FENG ; Na WANG
China Journal of Chinese Materia Medica 2007;32(13):1317-1319
OBJECTIVETo study the effects of resvaratrol derivatives on spontaneous HR and CF of isolated guinea pig atrium.
METHODThe dose-effect curve of resvaratrol was observed. The possible mechanism of potassium channels responsible for changes of CF and HR after administering with resvaratrol was measured.
RESULTResvaratrol reduced the spontaneous HR and weakened the CF in a dose-dependent manner ranging from 10(-6) to 3 x 10(-4) mol x L(-1) (P < 0.05). As compared with Res group, the effects were partly blocked by Gli (P < 0.05) and TEA (P < 0.01), but not blocked by 4-AP, BaCl2, Atropine.
CONCLUSIONResvaratrol can induce negative chronotropic action and negative (inotropic action. The mechanism(s) may relate to the opening of K(ATP) and Kc(Ca).
Animals ; Barium Compounds ; pharmacology ; Cardiotonic Agents ; administration & dosage ; isolation & purification ; pharmacology ; Chlorides ; pharmacology ; Dose-Response Relationship, Drug ; Female ; Glyburide ; pharmacology ; Guinea Pigs ; Heart Rate ; drug effects ; In Vitro Techniques ; KATP Channels ; antagonists & inhibitors ; Male ; Myocardial Contraction ; drug effects ; Plants, Medicinal ; chemistry ; Potassium Channel Blockers ; pharmacology ; Potassium Channels, Calcium-Activated ; antagonists & inhibitors ; Potassium Channels, Inwardly Rectifying ; antagonists & inhibitors ; Stilbenes ; administration & dosage ; isolation & purification ; pharmacology ; Tetraethylammonium ; pharmacology