Simultaneous all-optical manipulation and recording of neural circuit activity with cellular resolution in vivo.
Simultaneous all-optical manipulation and recording of neural circuit activity with cellular resolution in vivo.
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DOI：
10.1038/nmeth.3217
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发表时间：
2015-02
期刊：
影响因子：
48
通讯作者：
Häusser M
中科院分区：
文献类型：
--
作者：
Packer AM;Russell LE;Dalgleish HW;Häusser M
我们描述了一种全光学策略，用于同时操纵和记录多个神经元的活动与细胞分辨率在体内。通过共表达红移视蛋白和遗传编码的钙指示剂，能够实现同时的双光子光遗传学激活和钙成像。空间光调制器允许数十个用户选择的神经元被靶向用于时空精确的光遗传学激活，而同时快速钙成像提供具有可忽略的光学串扰的操纵的高分辨率网络范围的读出。在小鼠桶皮质中的原理证明实验证明了在不同的行为状态下对相同神经元群体的询问，以及基于其功能特征的神经元集合的靶向。这种方法将光遗传学工具包扩展到遗传或病毒方法获得的特异性之外，从而能够在体内哺乳动物大脑中以单细胞和单尖峰分辨率对功能定义的神经回路进行高通量、灵活和长期的光学询问。
We describe an all-optical strategy for simultaneously manipulating and recording the activity of multiple neurons with cellular resolution in vivo. Concurrent two-photon optogenetic activation and calcium imaging is enabled by coexpression of a red-shifted opsin and a genetically encoded calcium indicator. A spatial light modulator allows tens of user-selected neurons to be targeted for spatiotemporally precise optogenetic activation, while simultaneous fast calcium imaging provides high-resolution network-wide readout of the manipulation with negligible optical crosstalk. Proof-of-principle experiments in mouse barrel cortex demonstrate interrogation of the same neuronal population during different behavioral states, and targeting of neuronal ensembles based on their functional signature. This approach extends the optogenetic toolkit beyond the specificity obtained with genetic or viral approaches, enabling high-throughput, flexible and long-term optical interrogation of functionally defined neural circuits with single-cell and single-spike resolution in the mammalian brain in vivo.