美国斯坦福大学Michael Z. Lin与法国PSL研究大学Stéphane Dieudonné等研究人员合作开发了能够在清醒行为小鼠中进行超快双光子成像的电压指示蛋白。该研究于2019年12月12日发表于国际一流学术期刊《细胞》。

研究人员报告了ASAP3，这是一种遗传编码的电压指示蛋白，其通过生理电压、亚毫秒级激活动力学和双光子激发下的完全响应性具有51％的荧光变化。研究人员还介绍了一种用于体内千赫兹速率双光子采样的超快局部体积激发（ULoVE）方法，具有更高的稳定性和灵敏度。结合以体细胞为目标的ASAP3变体和ULoVE，研究人员在深处显示了几分钟的峰值和亚阈值事件的单次试验跟踪，这具有亚细胞分辨率，并能够在几天内重复采样。在视觉皮层中，研究人员使用针对性的ASAP3来说明运动引起的细胞类型依赖性亚阈值调节。因此，ASAP3和ULoVE能够在清醒行为期间在位于深处的遗传定义的神经元中高速光学记录电活动。

据介绍，用细胞分辨率光学研究大脑深部位置的电压，对于理解神经元回路如何处理信息将非常有用。

附：英文原文

Title: Ultrafast Two-Photon Imaging of a High-Gain Voltage Indicator in Awake Behaving Mice

Author: Vincent Villette, Mariya Chavarha, Ivan K. Dimov, Jonathan Bradley, Lagnajeet Pradhan, Benjamin Mathieu, Stephen W. Evans, Simon Chamberland, Dongqing Shi, Renzhi Yang, Benjamin B. Kim, Annick Ayon, Abdelali Jalil, Franois St-Pierre, Mark J. Schnitzer, Guoqiang Bi, Katalin Toth, Jun Ding, Stéphane Dieudonné, Michael Z. Lin

Issue&Volume: 2019/12/12

Abstract: Optical interrogation of voltage in deep brain locations with cellular resolutionwould be immensely useful for understanding how neuronal circuits process information.Here, we report ASAP3, a genetically encoded voltage indicator with 51% fluorescencemodulation by physiological voltages, submillisecond activation kinetics, and fullresponsivity under two-photon excitation. We also introduce an ultrafast local volumeexcitation (ULoVE) method for kilohertz-rate two-photon sampling in vivo with increased stability and sensitivity. Combining a soma-targeted ASAP3 variantand ULoVE, we show single-trial tracking of spikes and subthreshold events for minutesin deep locations, with subcellular resolution and with repeated sampling over days.In the visual cortex, we use soma-targeted ASAP3 to illustrate cell-type-dependentsubthreshold modulation by locomotion. Thus, ASAP3 and ULoVE enable high-speed opticalrecording of electrical activity in genetically defined neurons at deep locationsduring awake behavior.

DOI: 10.1016/j.cell.2019.11.004

Source: https://www.cell.com/cell/fulltext/S0092-8674(19)31225-5