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Automated Intracellular Pharmacological Electrophysiology for Ligand-Gated Ionotropic Receptor and Pharmacology Screening
Molecular Pharmacology ( IF 3.2 ) Pub Date : 2021-07-01 , DOI: 10.1124/molpharm.120.000195
Riley E Perszyk 1 , Mighten C Yip 1 , Ona L McConnell 1 , Eric T Wang 1 , Andrew Jenkins 2 , Stephen F Traynelis 2 , Craig R Forest 2
Affiliation  

Communication between neuronal cells, which is central to brain function, is performed by several classes of ligand-gated ionotropic receptors. The gold-standard technique for measuring rapid receptor response to agonist is manual patch-clamp electrophysiology, capable of the highest temporal resolution of any current electrophysiology technique. We report an automated high-precision patch-clamp system that substantially improves the throughput of these time-consuming pharmacological experiments. The patcherBotPharma enables recording from cells expressing receptors of interest and manipulation of them to enable millisecond solution exchange to activate ligand-gated ionotropic receptors. The solution-handling control allows for autonomous pharmacological concentration-response experimentation on adherent cells, lifted cells, or excised outside-out patches. The system can perform typical ligand-gated ionotropic receptor experimentation protocols autonomously, possessing a high success rate in completing experiments and up to a 10-fold reduction in research effort over the duration of the experiment. Using it, we could rapidly replicate previous data sets, reducing the time it took to produce an eight-point concentration-response curve of the effect of propofol on GABA type A receptor deactivation from likely weeks of recording to ∼13 hours of recording. On average, the rate of data collection of the patcherBotPharma was a data point every 2.1 minutes that the operator spent interacting with the patcherBotPharma. The patcherBotPharma provides the ability to conduct complex and comprehensive experimentation that yields data sets not normally within reach of conventional systems that rely on constant human control. This technical advance can contribute to accelerating the examination of the complex function of ion channels and the pharmacological agents that act on them.

中文翻译:

用于配体门控离子型受体和药理学筛选的自动化细胞内药理学电生理学

神经元细胞之间的通讯是大脑功能的核心,由几类配体门控离子型受体进行。测量受体对激动剂的快速反应的金标准技术是手动膜片钳电生理学,它能够实现任何当前电生理学技术的最高时间分辨率。我们报告了一种自动化的高精度膜片钳系统,它大大提高了这些耗时的药理学实验的吞吐量。patcherBot制药能够记录表达感兴趣受体的细胞并对其进行操作,以实现毫秒溶液交换以激活配体门控离子型受体。溶液处理控制允许对贴壁细胞、提升细胞或切除的外贴片进行自主药理学浓度反应实验。该系统可以自主执行典型的配体门控离子型受体实验方案,在完成实验方面具有很高的成功率,并且在实验期间可以将研究工作量减少多达 10 倍。使用它,我们可以快速复制以前的数据集,从而将产生异丙酚对 GABA A 型受体失活影响的八点浓度-反应曲线的时间从可能的记录周缩短到约 13 小时的记录。Pharma是操作员与 patcherBot Pharma交互所花费的每 2.1 分钟的一个数据点。patcherBot Pharma提供了进行复杂而全面的实验的能力,这些实验产生的数据集通常无法达到依赖于持续人工控制的传统系统的范围。这一技术进步有助于加速对离子通道的复杂功能和作用于它们的药物的检查。
更新日期:2021-07-12
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