Werley Christopher A, Brookings Ted, Upadhyay Hansini, Williams Luis A, McManus Owen B, Dempsey Graham T
Q-State Biosciences, Cambridge, Massachusetts.
Curr Protoc Pharmacol. 2017 Sep 11;78:11.20.1-11.20.24. doi: 10.1002/cpph.25.
A key challenge for establishing a phenotypic screen for neuronal excitability is measurement of membrane potential changes with high throughput and accuracy. Most approaches for probing excitability rely on low-throughput, invasive methods or lack cell-specific information. These limitations stimulated the development of novel strategies for characterizing the electrical properties of cultured neurons. Among these was the development of optogenetic technologies (Optopatch) that allow for stimulation and recording of membrane voltage signals from cultured neurons with single-cell sensitivity and millisecond temporal resolution. Neuronal activity is elicited using blue light activation of the channelrhodopsin variant 'CheRiff'. Action potentials and synaptic signals are measured with 'QuasAr', a rapid and sensitive voltage-indicating protein with near-infrared fluorescence that scales proportionately with transmembrane potential. This integrated technology of optical stimulation and recording of electrical signals enables investigation of neuronal electrical function with unprecedented scale and precision. © 2017 by John Wiley & Sons, Inc.
建立神经元兴奋性表型筛选的一个关键挑战是高通量且准确地测量膜电位变化。大多数探测兴奋性的方法依赖于低通量、侵入性方法，或者缺乏细胞特异性信息。这些局限性推动了用于表征培养神经元电特性的新策略的发展。其中包括光遗传学技术（Optopatch）的开发，该技术能够以单细胞灵敏度和毫秒级时间分辨率刺激并记录培养神经元的膜电压信号。使用蓝光激活通道视紫红质变体“CheRiff”来引发神经元活动。动作电位和突触信号通过“QuasAr”进行测量，“QuasAr”是一种快速且灵敏的具有近红外荧光的电压指示蛋白，其荧光强度与跨膜电位成比例变化。这种光刺激和电信号记录的集成技术能够以前所未有的规模和精度研究神经元的电功能。© 2017约翰威立国际出版公司
