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：

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时间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中以高时间精度对单个神经元进行光遗传学控制，将使对神经回路运作方式的探索成为可能。双光子计算机生成的全息术可以精确雕刻光，原则上可以同时照射网络中的许多神经元，具有必要的时间精度来模拟准确的神经代码。我们设计了一种高效的胞体靶向视蛋白，发现将红藻氨酸受体亚基 2 (KA2) 的 N 末端 150 个残基与最近发现的高光电流通道视紫红质 CoChR 融合，主要限制这种视蛋白在哺乳动物皮质细胞体中的表达神经元。结合双光子全息刺激，我们发现这种体细胞 CoChR (soCoChR) 能够以单细胞分辨率和 <1 毫秒的时间精度对小鼠皮质脑切片中的单个细胞进行光刺激。我们使用 soCoChR 在完整的皮质电路上执行连接映射。