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大鼠脑cDNA表达的钠通道的膜片钳特性分析

Patch clamp characterization of sodium channels expressed from rat brain cDNA.

作者信息

Stühmer W, Methfessel C, Sakmann B, Noda M, Numa S

出版信息

Eur Biophys J. 1987;14(3):131-8. doi: 10.1007/BF00253837.

Abstract

Sodium currents, INa, were recorded from Xenopus laevis oocytes which had been injected with mRNA synthesized by in vitro transcription of the rat brain sodium channel II cDNA (Noda et al. 1986 a, b). Patch pipettes were used to apply depolarizing voltage steps and to record macroscopic sodium currents of between 50 and 750 pA from cell-attached patches of the oocyte membrane. With a combination of whole-cell and patch clamp recording, the properties of the implanted sodium channels could be studied in detail. They were analyzed according to the model of Hodgkin and Huxley (1952 a) assuming three activation gates. The activation of the sodium currents is characterized by an equilibrium potential of -29 mV and an apparent gating charge of 8.7 e0. At -64 mV half of the sodium currents were inactivated. From single-channel current recordings, an elementary sodium channel conductance of 19 pS and an average open time of 0.43 ms were obtained at -32 mV membrane potential and 16 degrees C. The single-channel and activation properties of rat brain sodium channel II are therefore comparable to those found in peripheral nerve and skeletal muscle, but inactivation occurs at less negative potentials. This could be a specific property of the brain sodium channels and may underlie the maintained inward sodium currents reported in brain neurones (French and Gage 1985).

摘要

钠电流(INa)是从非洲爪蟾卵母细胞中记录的,这些卵母细胞已注射了通过大鼠脑钠通道II cDNA的体外转录合成的mRNA(野田等人,1986年a,b)。膜片钳吸管用于施加去极化电压阶跃,并记录来自卵母细胞膜贴附式膜片的50至750 pA的宏观钠电流。结合全细胞和膜片钳记录,可以详细研究植入的钠通道的特性。根据霍奇金和赫胥黎(1952年a)的模型,假设存在三个激活门,对它们进行了分析。钠电流的激活特征在于平衡电位为-29 mV,表观门控电荷为8.7 e0。在-64 mV时,一半的钠电流失活。从单通道电流记录中,在-32 mV膜电位和16摄氏度下,获得了19 pS的基本钠通道电导和0.43 ms的平均开放时间。因此,大鼠脑钠通道II的单通道和激活特性与在周围神经和骨骼肌中发现的特性相当,但失活发生在更正的电位。这可能是脑钠通道的一个特定特性,可能是脑神经元中报道的持续内向钠电流的基础(弗伦奇和盖奇,1985年)。

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