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人类红细胞中的酸敏感外向整流阴离子通道。

Acid-sensitive outwardly rectifying anion channels in human erythrocytes.

作者信息

Kucherenko Yuliya V, Mörsdorf Daniel, Lang Florian

机构信息

Department of Physiology, University of Tübingen, Tübingen, Germany.

出版信息

J Membr Biol. 2009 Jul;230(1):1-10. doi: 10.1007/s00232-009-9179-z. Epub 2009 Jul 2.

Abstract

Acid-sensitive outwardly rectifying anion channels (ASOR) have been described in several mammalian cell types. The present whole-cell patch-clamp study elucidated whether those channels are expressed in erythrocytes. To this end whole-cell recordings were made in human erythrocytes from healthy donors treated with low pH and high osmotic pressure. When the pipette solution had a reduced Cl(-) concentration, treatment of the cells with Cl(-)-containing normal and hyperosmotic (addition of sucrose and polyethelene glycol 1000 [PEG-1000] to the Ringer) media with low pH significantly increased the conductance of the cells at positive voltages. Channel activity was highest in the PEG-1000 media (95 and 300 mM PEG-1000, pH 4.5 and 4.3, respectively) where the current-voltage curves demonstrated strong outward rectification and reversed at -40 mV. Substitution of the Cl(-)-containing medium with Cl(-)-free medium resulted in a decrease of the conductance at hyperpolarizing voltages, a shift in reversal potential (to 0 mV) and loss of outward rectification. The chloride currents were inhibited by chloride channels blockers DIDS and NPPB (IC(50) for both was approximately 1 mM) but not with niflumic acid and amiloride. The observations reveal expression of ASOR in erythrocytes.

摘要

酸敏感外向整流阴离子通道(ASOR)已在多种哺乳动物细胞类型中被描述。本全细胞膜片钳研究阐明了这些通道是否在红细胞中表达。为此,对来自健康供体的人类红细胞进行全细胞记录,这些红细胞用低pH和高渗透压处理。当移液管溶液中的Cl⁻浓度降低时,用含Cl⁻的正常和高渗(向林格液中添加蔗糖和聚乙二醇1000 [PEG - 1000])低pH培养基处理细胞,在正电压下显著增加了细胞的电导。通道活性在PEG - 1000培养基(分别为95和300 mM PEG - 1000,pH 4.5和4.3)中最高,此时电流 - 电压曲线显示出强烈的外向整流,并在 - 40 mV处反转。用无Cl⁻培养基替代含Cl⁻培养基导致超极化电压下电导降低、反转电位偏移(至0 mV)以及外向整流消失。氯电流受到氯通道阻滞剂DIDS和NPPB的抑制(两者的IC₅₀约为1 mM),但不受氟尼酸和阿米洛利的影响。这些观察结果揭示了红细胞中ASOR的表达。

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