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通过在非洲爪蟾胚胎细胞中表达外源钾通道基因对当前特性进行修饰。

Modifications of current properties by expression of a foreign potassium channel gene in Xenopus embryonic cells.

作者信息

Spruce A E, Moody W J

机构信息

Department of Pharmacology, Medical School, University of Birmingham, Edgbaston, UK.

出版信息

J Membr Biol. 1995 Dec;148(3):255-62. doi: 10.1007/BF00235043.

Abstract

The development of excitable cells is characterized by highly organized patterns of expression of ion channels. During the terminal differentiation of Xenopus muscle somites, potassium currents are expressed first just after Stage 15 (early-mid neurula), following a long period during which no voltage-dependent currents can be detected in any cell in the dorsal embryo. We have investigated whether early expression of a foreign delayed rectifier potassium channel may affect this endogenous pattern of electrical development. We injected the purified cRNA of the mammalian brain Shaker-like potassium channel, Kv1.1, into fertilized Xenopus eggs. The resulting currents were analyzed in blastomeres during a 12-hr period prior to Stage 15 and in differentiating muscle cells after Stage 15. In injected embryos, a high fraction of blastomeres expressed a delayed rectifier-type current. The Kv1.1 current could be distinguished from the endogenous muscle delayed potassium current (IK,X) by its very different voltage dependence. Separation of currents based on this difference indicated that, in injected embryos, IK,X appeared much earlier in development than in control embryos. Furthermore, even in cells which expressed solely Kv1.1-type current, the sensitivity of the current to dendrotoxin declined dramatically during development, approaching that of IK,X. These data suggest an interaction between Kv1.1 and endogenous channel subunits, and/or modification of the Kv1.1 protein by the embryonic cells in ways not seen in Xenopus oocytes or mammalian cell lines.

摘要

可兴奋细胞的发育特征是离子通道表达具有高度组织化的模式。在非洲爪蟾肌肉体节的终末分化过程中,钾电流在第15阶段(神经胚早期至中期)刚结束后首先表达,在此之前的很长一段时间里,背侧胚胎中的任何细胞都检测不到电压依赖性电流。我们研究了外源延迟整流钾通道的早期表达是否会影响这种内源性电发育模式。我们将哺乳动物脑Shaker样钾通道Kv1.1的纯化cRNA注射到非洲爪蟾受精卵中。在第15阶段之前的12小时内对卵裂球中的电流进行分析,并在第15阶段之后对分化中的肌肉细胞进行分析。在注射后的胚胎中,很大一部分卵裂球表达了延迟整流型电流。Kv1.1电流可以通过其非常不同的电压依赖性与内源性肌肉延迟钾电流(IK,X)区分开来。基于这种差异对电流进行分离表明,在注射后的胚胎中,IK,X在发育中出现的时间比对照胚胎早得多。此外,即使在仅表达Kv1.1型电流的细胞中,该电流对树突毒素的敏感性在发育过程中也显著下降,接近IK,X的敏感性。这些数据表明Kv1.1与内源性通道亚基之间存在相互作用,和/或胚胎细胞以非洲爪蟾卵母细胞或哺乳动物细胞系中未观察到的方式对Kv1.1蛋白进行修饰。

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