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Temporally precise single-cell-resolution optogenetics
Nature Neuroscience ( IF 25.0 ) Pub Date : 2017-11-13 , DOI: 10.1038/s41593-017-0018-8
Or A. Shemesh,Dimitrii Tanese,Valeria Zampini,Changyang Linghu,Kiryl Piatkevich,Emiliano Ronzitti,Eirini Papagiakoumou,Edward S. Boyden,Valentina Emiliani

Optogenetic control of individual neurons with high temporal precision within intact mammalian brain circuitry would enable powerful explorations of how neural circuits operate. Two-photon computer-generated holography enables precise sculpting of light and could in principle enable simultaneous illumination of many neurons in a network, with the requisite temporal precision to simulate accurate neural codes. We designed a high-efficacy soma-targeted opsin, finding that fusing the N-terminal 150 residues of kainate receptor subunit 2 (KA2) to the recently discovered high-photocurrent channelrhodopsin CoChR restricted expression of this opsin primarily to the cell body of mammalian cortical neurons. In combination with two-photon holographic stimulation, we found that this somatic CoChR (soCoChR) enabled photostimulation of individual cells in mouse cortical brain slices with single-cell resolution and <1-ms temporal precision. We used soCoChR to perform connectivity mapping on intact cortical circuits.

中文翻译:

暂时精确的单细胞分辨率光遗传学

在完整的哺乳动物大脑回路内以高的时间精度对单个神经元进行光遗传学控制,将使人们有力地探索神经回路的运作方式。两光子计算机生成的全息照相技术可以精确雕刻光,并且原则上可以同时照亮网络中的许多神经元,同时具有必要的时间精度以模拟精确的神经代码。我们设计了一种高效的针对体细胞的视蛋白,发现将红藻氨酸受体亚基2(KA2)的N末端150个残基融合到最近发现的高光电流通道视紫红质CoChR中,使该视蛋白的表达主要限制在哺乳动物皮质的细胞体中神经元。结合两光子全息刺激,我们发现,这种体细胞CoChR(soCoChR)能够以单细胞分辨率和<1毫秒时间精度对小鼠皮质大脑切片中的单个细胞进行光刺激。我们使用soCoChR在完整的皮层电路上执行连接映射。
更新日期:2017-11-13
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