Bright and photostable chemigenetic indicators for extended in vivo voltage imaging
Bright and photostable chemigenetic indicators for extended in vivo voltage imaging
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DOI：
10.1126/science.aav6416
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发表时间：
2019-08-16
期刊：
影响因子：
56.9
通讯作者：
Schreiter, Eric R.
中科院分区：
文献类型：
--
作者：
Abdelfattah, Ahmed S.;Kawashima, Takashi;Schreiter, Eric R.
基因编码电压指示器（GEVIs）能够在高空间和时间分辨率下监测神经元活动。然而，现有的gevi的应用受到荧光蛋白和视紫红质的亮度和光稳定性的限制。我们设计了一种名为Voltron的GEVI，它使用明亮且光稳定的合成染料代替基于蛋白质的荧光团，从而将体内同时成像的神经元数量增加了10倍，并且相对于现有的GEVI，能够成像的持续时间明显更长。我们使用Voltron对小鼠、斑马鱼和果蝇进行体内电压成像。在小鼠皮层中，Voltron允许在15分钟的连续成像期间同时记录来自数十个神经元的峰值和阈下电压信号。在斑马鱼幼体中，Voltron使刺突时间与行为精确相关。
Genetically encoded voltage indicators (GEVIs) enable monitoring of neuronal activity at high spatial and temporal resolution. However, the utility of existing GEVIs has been limited by the brightness and photostability of fluorescent proteins and rhodopsins. We engineered a GEVI, called Voltron, that uses bright and photostable synthetic dyes instead of protein-based fluorophores, thereby extending the number of neurons imaged simultaneously in vivo by a factor of 10 and enabling imaging for significantly longer durations relative to existing GEVIs. We used Voltron for in vivo voltage imaging in mice, zebrafish, and fruit flies. In the mouse cortex, Voltron allowed single-trial recording of spikes and subthreshold voltage signals from dozens of neurons simultaneously over a 15-minute period of continuous imaging. In larval zebrafish, Voltron enabled the precise correlation of spike timing with behavior.