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首页> 外文期刊>Neuroscience Letters: An International Multidisciplinary Journal Devoted to the Rapid Publication of Basic Research in the Brain Sciences >Human embryonic kidney (HEK293) cells express endogenous voltage-gated sodium currents and Na v 1.7 sodium channels.
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Human embryonic kidney (HEK293) cells express endogenous voltage-gated sodium currents and Na v 1.7 sodium channels.

机译：人类胚胎肾脏（HEK293）细胞表达内源性电压门控钠电流和Na v 1.7钠通道。

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摘要

Human embryonic kidney (HEK293) cells are widely used for the heterologous expression of voltage- and ligand-gated ion channels. Patch clamp analysis of HEK293 cells in the whole-cell configuration identified voltage-gated, rapidly inactivating inward currents. Peak current amplitudes ranged from less than 100 pA to more than 800 pA, with the majority (84 of 130 cells) in the 100-400 pA range. Transient inward currents were separated into three components on the basis of sensitivity to cadmium and tetrodotoxin (TTX). Application of cadmium (300 microM) reduced current amplitude to 65% of control, consistent with the existence of current carried by a cadmium-sensitive nonspecific cation channel previously identified in HEK293 cells. Application of TTX (500 nM) reduced current amplitude by 47%, consistent with the existence of current carried by a TTX-sensitive voltage-gated sodium channel. Joint application of cadmium and TTX was additive, reducing current amplitude to 28% of control. The residual cadmium- and TTX-resistant currents represent a third pharmacologically distinct component of the rapidly inactivating inward current that was not characterized further. The pyrethroid insecticide tefluthrin (10 microM) prolonged the inactivation of transient currents and induced slowly decaying tail currents, effects that are characteristic of sodium channel modification by pyrethroids. The use of sodium channel isoform-specific primers in polymerase chain reaction amplifications on HEK293 cell first-strand cDNA detected the consistent expression of the human Na(v)1.7 sodium channel isoform in cells that expressed the TTX-sensitive component of current. These results provide evidence for an endogenous TTX-sensitive sodium current in HEK293 cells that is associated primarily with the expression of the Na(v)1.7 sodium channel isoform.
机译：人类胚胎肾脏（HEK293）细胞被广泛用于电压和配体门控离子通道的异源表达。全细胞配置的HEK293细胞的膜片钳分析确定了电压门控的，迅速失活的内向电流。峰值电流幅度范围从小于100 pA到大于800 pA，大多数（130个电池中的84个）在100-400 pA范围内。根据对镉和河豚毒素（TTX）的敏感性，将瞬态内向电流分为三个部分。镉（300 microM）的施加将电流幅度减小到控制的65％，这与先前在HEK293细胞中鉴定出的对镉敏感的非特异性阳离子通道所载电流的存在相一致。 TTX（500 nM）的应用将电流幅度降低了47％，这与TTX敏感的电压门控钠通道所承载的电流的存在相一致。镉和TTX的联合应用是可加性的，可将电流幅度降低至对照组的28％。残留的抗镉和TTX的电流代表了快速失活的内向电流的第三个药理学独特成分，目前尚未进一步表征。拟除虫菊酯杀虫剂tefluthrin（10 microM）延长了瞬态电流的灭活并诱导了缓慢衰减的尾电流，这是拟除虫菊酯修饰钠通道的特征。在HEK293细胞第一链cDNA上的聚合酶链反应扩增中使用钠通道亚型特异性引物可检测出表达电流的TTX敏感成分的细胞中人Na（v）1.7钠通道亚型的一致表达。这些结果为HEK293细胞内源性TTX敏感性钠电流提供了证据，该电流主要与Na（v）1.7钠通道亚型的表达有关。

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