A new electro-optical approach for conductance measurement: an assay for the study of drugs acting on ligand-gated ion channels
A new electro-optical approach for conductance measurement: an assay for the study of drugs acting on ligand-gated ion channels
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一种新的电导测量电光方法：研究作用于配体门控离子通道的药物的测定方法
作者：
A. Menegon;S. Pitassi;N. Mazzocchi;L. Redaelli;R. Rizzetto;J. Rolland;C. Poli;M. Imberti;A. Lanati;F. Grohovaz
配体门控离子通道参与了许多病理生理过程，并代表了一个相关的，但具有挑战性的药物发现的目标。我们提出了一种创新的电光方法，他们的分析能够获得膜电导值从局部膜电位的变化所施加的测试电流脉冲和快速电压敏感的荧光染料测量。我们开发了一种名为“ionChannel光学高含量显微镜”（ionChannelΩ）的药物检测系统，从而充分利用了这种专有方法的潜力。通过测试参比药物对表达不同配体门控离子通道的细胞的响应来验证该自动化平台。此外，FLIPR和自动膜片钳的双盲比较进行设计作为拮抗剂的P2RX7受体的分子。ionChannelΩ在所有测试中均被证明具有高度可靠性，比电生理技术更快、更具成本效益。总体而言，ionChannelΩ适合于研究配体门控离子通道，由于当前测定的局限性，配体门控离子通道受到较少关注。
Ligand gated ion channels are involved in many pathophysiological processes and represent a relevant, although challenging, target for drug discovery. We propose an innovative electro-optical approach to their analysis able to derive membrane conductance values from the local membrane potential changes imposed by test current pulses and measured by fast voltage-sensitive fluorescent dyes. We exploited the potential of this proprietary method by developing a drug testing system called “ionChannel Optical High-content Microscope” (ionChannelΩ). This automated platform was validated by testing the responses of reference drugs on cells expressing different ligand-gated ion channels. Furthermore, a double-blind comparison with FLIPR and automated patch-clamp was performed on molecules designed to act as antagonists of the P2RX7 receptor. ionChannelΩ proved highly reliable in all tests, resulting faster and more cost-effective than electrophysiological techniques. Overall, ionChannelΩ is amenable to the study of ligand gated ion channels that are receiving less attention due to limitations in current assays.