Modulation by Divalent Cations of GABAρ1 Receptor From Human Retina Expressed in Xenopus Oocytes

WANG Qin-wen; WANG FENG; YANG JI-QING; LI ZHI-WANG

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Volume 14 Issue 4

Dec. 2001

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Article Navigation > Biomedical and Environmental Sciences > 2001 > 14(4): 269-277

WANG Qin-wen, WANG FENG, YANG JI-QING, LI ZHI-WANG. Modulation by Divalent Cations of GABAρ1 Receptor From Human Retina Expressed in Xenopus Oocytes[J]. Biomedical and Environmental Sciences, 2001, 14(4): 269-277.

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WANG Qin-wen, WANG FENG, YANG JI-QING, LI ZHI-WANG. Modulation by Divalent Cations of GABAρ1 Receptor From Human Retina Expressed in Xenopus Oocytes[J]. Biomedical and Environmental Sciences, 2001, 14(4): 269-277.

Modulation by Divalent Cations of GABAρ1 Receptor From Human Retina Expressed in Xenopus Oocytes

Research Center of Basic Medicine, Wuhan Institute of Children Disease,

Abstract

Objective To investigate functional homooligomeric GABAρ1 receptors expressed in Xenopus oocytes and the modulation of divalent cations. Methods　GABAρ1 cDNA from human retina was transcribed in vitro to obtain sense ρ1 mRNA, which was microinjected into Xenopus oocytes. Two-electrodes voltage clamp technique was performed to record GABA-induced currents. Results　 Expressed receptors were found to have similar properties to GABAC receptors characterized in the retina. Cl-currents induced by GABA were blocked by picrotoxin instead of bicuculline. GABA-induced currents reversed at -19±2.5 mV, and EC50 was 3.3 μmol/L. Zn+　+ modulated GABA-induced currents with an IC50=9.6 μ mol/L. Ni+　+, Cu+　+ and Cd+　+ inhibited GABA ρ1 obviously, too. Their rank order of potency was Zn+　+>Ni+　+>Cu+　+>Cd+　+. Conclusion　Zinc(10　μmol/L) inhibited GABA-induced currents in a competitive manner, and its action was sensitive to extracellular pH. Site-directed mutagenesis revealed that substitution of a single histidine residue (H44 and H48) failed to affect zinc sensitivity.
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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

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Modulation by Divalent Cations of GABAρ1 Receptor From Human Retina Expressed in Xenopus Oocytes

Research Center of Basic Medicine, Wuhan Institute of Children Disease,

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instead of /
in vitro

Abstract: Objective To investigate functional homooligomeric GABAρ1 receptors expressed in Xenopus oocytes and the modulation of divalent cations. Methods　GABAρ1 cDNA from human retina was transcribed in vitro to obtain sense ρ1 mRNA, which was microinjected into Xenopus oocytes. Two-electrodes voltage clamp technique was performed to record GABA-induced currents. Results　 Expressed receptors were found to have similar properties to GABAC receptors characterized in the retina. Cl-currents induced by GABA were blocked by picrotoxin instead of bicuculline. GABA-induced currents reversed at -19±2.5 mV, and EC50 was 3.3 μmol/L. Zn+　+ modulated GABA-induced currents with an IC50=9.6 μ mol/L. Ni+　+, Cu+　+ and Cd+　+ inhibited GABA ρ1 obviously, too. Their rank order of potency was Zn+　+>Ni+　+>Cu+　+>Cd+　+. Conclusion　Zinc(10　μmol/L) inhibited GABA-induced currents in a competitive manner, and its action was sensitive to extracellular pH. Site-directed mutagenesis revealed that substitution of a single histidine residue (H44 and H48) failed to affect zinc sensitivity.

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