Evaluation of toxicity equivalent factors of paralytic shellfish poisoning toxins in seven human sodium channels types by an automated high throughput electrophysiology system
Evaluation of toxicity equivalent factors of paralytic shellfish poisoning toxins in seven human sodium channels types by an automated high throughput electrophysiology system
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DOI：
10.1007/s00204-014-1444-y
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发表时间：
2016-02-01
影响因子：
6.1
通讯作者：
Botana, Luis M.
中科院分区：
文献类型：
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作者：
Alonso, Eva;Alfonso, Amparo;Botana, Luis M.
尽管电压门控钠通道（Na（v））是麻痹性贝类毒素（PSP）的细胞靶点，膜片钳电生理学是研究分子与这些通道直接相互作用的最有效方法，但由于难以将其转化为可靠的通量系统，目前该技术仍被简化为更具体的分析。PSP检测的实际功能方法基于使用受体而不是功能性Na（v）通道的结合测定。目前，自动化膜片钳平台的可用性，也与人类Na（V）通道的稳定转染细胞系，使我们能够引入这种特定的和选择性的方法，快速筛选海洋毒素检测。利用可获得的纯PSP标准品的优势，我们计算了9个PSP类似物的毒性等效因子（TEFs），获得了可靠的TEFs在人类目标中，以弥补官方分析方法的不足，并验证自动膜片钳技术作为一种快速和可靠的筛选方法与钠通道相互作用的海洋毒素。这项工作的主要观察结果是TEF的变化很大，这取决于所选择的通道亚型，值得注意的是1.7通道亚型的效力变化以及Na（v）1.6和1.2通道对PSP筛选的适用性。
Although voltage-gated sodium channels (Na (v) ) are the cellular target of paralytic shellfish poisoning (PSP) toxins and that patch clamp electrophysiology is the most effective way of studying direct interaction of molecules with these channels, nowadays, this technique is still reduced to more specific analysis due to the difficulties of transforming it in a reliable throughput system. Actual functional methods for PSP detection are based in binding assays using receptors but not functional Na (v) channels. Currently, the availability of automated patch clamp platforms and also of stably transfected cell lines with human Na (v) channels allow us to introduce this specific and selective method for fast screenings in marine toxin detection. Taking advantage of the accessibility to pure PSP standards, we calculated the toxicity equivalent factors (TEFs) for nine PSP analogs obtaining reliable TEFs in human targets to fulfill the deficiencies of the official analytic methods and to verify automated patch clamp technology as a fast and reliable screening method for marine toxins that interact with the sodium channel. The main observation of this work was the large variation of TEFs depending on the channel subtype selected, being remarkable the variation of potency in the 1.7 channel subtype and the suitability of Na (v) 1.6 and 1.2 channels for PSP screening.