San Raffaele Scientific Institute, via Olgettina 60, 20132, Milan, Italy.
San Raffaele University, via Olgettina 58, 20132, Milan, Italy.
Sci Rep. 2017 Mar 21;7:44843. doi: 10.1038/srep44843.
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.
配体门控离子通道参与许多病理生理过程，是药物发现的一个相关但具有挑战性的目标。我们提出了一种创新的光电分析方法，能够从测试电流脉冲引起的局部膜电位变化中得出膜电导值，并通过快速电压敏感荧光染料进行测量。我们通过开发一种名为“离子通道光学高内涵显微镜（ionChannelΩ）”的药物测试系统来利用这种专利方法的潜力。该自动化平台通过测试在表达不同配体门控离子通道的细胞上参考药物的反应进行了验证。此外，还与 FLIPR 和自动化膜片钳技术进行了双盲比较，比较的分子设计为 P2RX7 受体拮抗剂。ionChannelΩ 在所有测试中都非常可靠，比电生理技术更快、更具成本效益。总的来说，ionChannelΩ适用于研究由于当前检测方法的限制而受到较少关注的配体门控离子通道。
