Comparison of the pharmacological properties of rat Na(V)1.8 with rat Na(V)1.2a and human Na(V)1.5 voltage-gated sodium channel subtypes using a membrane potential sensitive dye and FLIPR.

Receptors & channels Pub Date : 2004-01-01 DOI:10.3109/10606820490270410

R. Vickery, S. Amagasu, R. Chang, N. Mai, E. Kaufman, J. Martin, J. Hembrador, M. O'Keefe, C. Gee, D. Marquess, Jacqueline A.M. Smith

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引用次数: 30

Abstract

A novel, membrane potential sensitive dye and a fluorescence imaging plate reader (FLIPR) have been used to characterize the pharmacological properties of rat Na(v)1.8 voltage-gated sodium channels (VGSC) in parallel with rat Na(v)1.2a and human Na(v)1.5 VGSC subtypes, respectively. The sensitivity of recombinant Na(v)1.2a-CHO, Na(v)1.5-293-EBNA, and Na(v)1.8-F-11 cells to VGSC activators was subtype dependent. Veratridine evoked depolarization of Na(v)1.2a-CHO and Na(v)1.5-293-EBNA cells with pEC(50) values of 4.78 +/- 0.13 and 4.84 +/- 0.12, respectively (n = 3), but had negligible effect on Na(v)1.8-F-11 cells (pEC(50) < 4.5). Type I pyrethroids were without significant effect at all subtypes. In contrast, the type II pyrethroids deltamethrin and fenvalerate evoked direct depolarization of Na(v)1.8-F-11 and Na(v)1.5-293-EBNA cells. Deltamethrin potentiated the veratridine-evoked response in Na(v)1.8-F-11 cells by > or =20-fold, in contrast to a

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利用膜电位敏感染料和FLIPR比较大鼠Na(V)1.8与大鼠Na(V)1.2a和人Na(V)1.5电压门控钠通道亚型的药理学性质。

利用一种新型的膜电位敏感染料和荧光成像板阅读器(FLIPR)分别表征了大鼠Na(v)1.8电压门控钠通道(VGSC)与大鼠Na(v)1.2a和人Na(v)1.5 VGSC亚型的药理学性质。重组Na(v)1.2a-CHO、Na(v)1.5-293-EBNA和Na(v)1.8-F-11细胞对VGSC激活剂的敏感性呈亚型依赖性。Veratridine对Na(v)1.2a-CHO和Na(v)1.5-293-EBNA细胞的去极化作用分别为pEC(50)值4.78 +/- 0.13和4.84 +/- 0.12 (n = 3)，对Na(v)1.8-F-11细胞的去极化作用可忽略(pEC(50) < 4.5)。I型拟除虫菊酯对所有亚型均无显著影响。II型拟除虫菊酯类杀虫剂溴氰菊酯和氰戊菊酯诱导Na(v)1.8-F-11和Na(v)1.5-293-EBNA细胞直接去极化。溴氰菊酯能使Na(v)1.8-F-11细胞的缬草碱诱发反应增强>或=20倍，而Na(v)1.2a和Na(v)1.5细胞的缬草碱诱发反应增强<或=3倍。河豚毒素(TTX)对VGSC激活剂诱导的Na(v)1.8-F-11细胞去极化有抑制作用，呈双相浓度-反应曲线。计算得到的pIC(50)值分别为8.05 +/- 0.25 (n = 4)和4.32 +/- 0.21 (n = 4)，对应于TTX对内源性TTX敏感(TTX- s)和重组Na(v)1.8 TTX耐药(TTX- r) VGSCs的抑制作用。除TTX外，许多离子通道阻滞剂对Na(v)1.8、Na(v)1.2a和Na(v)1.5 VGSC亚型的效价相似。综上所述，这些高通量FLIPR检测是测定不同VGSC亚型化合物相对效力的有价值的工具，并可能被证明对鉴定新的亚型选择性抑制剂有用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Receptors & channels

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