Huang Chien-Jung, Harootunian Alec, Maher Michael P, Quan Catherine, Raj Christopher D, McCormack Ken, Numann Randal, Negulescu Paul A, González Jesús E
Vertex Pharmaceuticals Incorporated, 11010 Torreyana Road, San Diego, California 92121, USA.
Nat Biotechnol. 2006 Apr;24(4):439-46. doi: 10.1038/nbt1194. Epub 2006 Mar 19.
Voltage-gated ion channels regulate many physiological functions and are targets for a number of drugs. Patch-clamp electrophysiology is the standard method for measuring channel activity because it fulfils the requirements for voltage control, repetitive stimulation and high temporal resolution, but it is laborious and costly. Here we report an electro-optical technology and automated instrument, called the electrical stimulation voltage ion probe reader (E-VIPR), that measures the activity of voltage-gated ion channels using extracellular electrical field stimulation and voltage-sensitive fluorescent probes. We demonstrate that E-VIPR can sensitively detect drug potency and mechanism of block on the neuronal human type III voltage-gated sodium channel expressed in human embryonic kidney cells. Results are compared with voltage-clamp and show that E-VIPR provides sensitive and information-rich compound blocking activity. Furthermore, we screened approximately 400 drugs and observed sodium channel-blocking activity for approximately 25% of them, including the antidepressants sertraline (Zoloft) and paroxetine (Paxil).
电压门控离子通道调节多种生理功能，是许多药物的作用靶点。膜片钳电生理学是测量通道活性的标准方法，因为它满足电压控制、重复刺激和高时间分辨率的要求，但操作繁琐且成本高昂。在此，我们报告了一种电光技术和自动化仪器，称为电刺激电压离子探针读数器（E-VIPR），它使用细胞外电场刺激和电压敏感荧光探针来测量电压门控离子通道的活性。我们证明，E-VIPR能够灵敏地检测药物对人胚胎肾细胞中表达的神经元人III型电压门控钠通道的效力和阻断机制。将结果与电压钳进行比较，表明E-VIPR提供了灵敏且信息丰富的化合物阻断活性。此外，我们筛选了约400种药物，观察到其中约25%具有钠通道阻断活性，包括抗抑郁药舍曲林（左洛复）和帕罗西汀（帕罗西汀）。
