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A6细胞的基底外侧膜钾离子电导

Basolateral membrane potassium conductance of A6 cells.

作者信息

Broillet M C, Horisberger J D

机构信息

Institut de Pharmacologie, Université de Lausanne, Switzerland.

出版信息

J Membr Biol. 1991 Oct;124(1):1-12. doi: 10.1007/BF01871359.

Abstract

To study the properties of the basolateral membrane conductance of an amphibian epithelial cell line, we have adapted the technique of apical membrane selective permeabilization (Wills, N.K., Lewis, S.A., Eaton, D.C. 1979b, J. Membrane Biol. 45:81-108). Monolayers of A6 cells cultured on permeable supports were exposed to amphotericin B. The apical membrane was effectively permeabilized, while the high electrical resistance of the tight junctions and the ionic selectivity of the basolateral membrane were preserved. Thus the transepithelial current-voltage relation reflected mostly the properties of the basolateral membrane. Under "basal" conditions, the basolateral membrane conductance was inward rectifying, highly sensitive to barium but not to quinidine. After the induction of cell swelling either by adding chloride to the apical solution or by lowering the osmolarity of the basolateral solution, a large outward-rectifying K+ conductance was observed, and addition of barium or quinidine to the basolateral side inhibited, respectively, 82.4 +/- 1.9% and 90.9 +/- 1.0% of the transepithelial current at 0 mV. Barium block was voltage dependent; the half-inhibition constant (Ki) varied from 1499 +/- 97 microM at 0 mV to 5.7 +/- 0.5 microM at -120 mV. Cell swelling induces a large quinidine-sensitive K+ conductance, changing the inward-rectifying basolateral membrane conductance observed under "basal" conditions into a conductance with outward-rectifying properties.

摘要

为了研究两栖动物上皮细胞系基底外侧膜电导的特性,我们采用了顶膜选择性通透技术(Wills, N.K., Lewis, S.A., Eaton, D.C. 1979b, J. Membrane Biol. 45:81 - 108)。将培养在可渗透支持物上的A6细胞单层暴露于两性霉素B。顶膜被有效通透,而紧密连接的高电阻和基底外侧膜的离子选择性得以保留。因此,跨上皮电流 - 电压关系主要反映了基底外侧膜的特性。在“基础”条件下,基底外侧膜电导呈内向整流,对钡高度敏感,但对奎尼丁不敏感。在通过向顶侧溶液中添加氯离子或降低基底外侧溶液的渗透压诱导细胞肿胀后,观察到一个大的外向整流钾电导,并且向基底外侧添加钡或奎尼丁分别在0 mV时抑制了82.4±1.9%和90.9±1.0%的跨上皮电流。钡阻断具有电压依赖性;半抑制常数(Ki)在0 mV时为1499±97 μM,在 - 120 mV时为5.7±0.5 μM。细胞肿胀诱导出一个对奎尼丁敏感的大钾电导,将“基础”条件下观察到的内向整流基底外侧膜电导转变为具有外向整流特性的电导。

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