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一种克隆成骨细胞样细胞系的电生理学:钙激活钾通道存在的证据。

Electrophysiology of a clonal osteoblast-like cell line: evidence for the existence of a Ca2+-activated K+ conductance.

作者信息

Dixon S J, Aubin J E, Dainty J

出版信息

J Membr Biol. 1984;80(1):49-58. doi: 10.1007/BF01868689.

Abstract

Intracellular microelectrode measurements were made on a well-characterized osteoblast-like clonal cell line isolated from a rat osteosarcoma. In serum-free medium, stable membrane potentials of -42 +/- 9 mV (SD, n = 190) were recorded. Ion substitution experiments suggested that this membrane potential is primarily a Na+/K+ diffusion potential. Input resistance was correlated strongly with colony size, ranging from 49 +/- 18 M omega (SD, n = 14) for colonies of 1-3 cells, to 4 +/- 4 M omega (SD, n = 164) for colonies of 100 or more cells. These results are consistent with the existence of low resistance intercellular junctions. Application of the carboxylic calcium ionophore A23187 by pressure microejection onto the cell surface resulted in a transient hyperpolarization and concomitant decrease in input resistance. Both these effects are consistent with an increased K+ conductance. Ion substitution experiments demonstrated that the degree of hyperpolarization was dependent on the external concentration of both K+ and Ca2+. Quinine, a blocker of Ca2+-activated K+ channels, inhibited the ionophore-induced hyperpolarization in a dose-dependent manner. It was concluded that these cells exhibit a Ca2+-activated K+ conductance.

摘要

在从大鼠骨肉瘤分离出的一种特征明确的成骨细胞样克隆细胞系上进行了细胞内微电极测量。在无血清培养基中,记录到稳定的膜电位为-42±9mV(标准差,n = 190)。离子置换实验表明,这种膜电位主要是一种Na+/K+扩散电位。输入电阻与集落大小密切相关,1 - 3个细胞的集落输入电阻为49±18MΩ(标准差,n = 14),而100个或更多细胞的集落输入电阻为4±4MΩ(标准差,n = 164)。这些结果与低电阻细胞间连接的存在相一致。通过压力微喷射将羧酸钙离子载体A23187施加到细胞表面,导致瞬时超极化并伴随输入电阻降低。这两种效应都与K+电导增加相一致。离子置换实验表明,超极化程度取决于细胞外K+和Ca2+的浓度。奎宁是一种Ca2+激活的K+通道阻滞剂,它以剂量依赖的方式抑制离子载体诱导的超极化。得出的结论是,这些细胞表现出Ca2+激活的K+电导。

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