An engineered channelrhodopsin optimized for axon terminal activation and circuit mapping.
An engineered channelrhodopsin optimized for axon terminal activation and circuit mapping.
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DOI：
10.1038/s42003-021-01977-7
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发表时间：
2021-04-12
影响因子：
5.9
通讯作者：
Ohtsuka T
中科院分区：
文献类型：
--
作者：
Hamada S;Nagase M;Yoshizawa T;Hagiwara A;Isomura Y;Watabe AM;Ohtsuka T
视紫红质通道蛋白 2 (ChR2) 等光遗传学工具可以对神经回路进行操作和绘制。然而，仍然缺乏选择性地沿着神经元的长距离轴突投射进行精确突触前激活的 ChR2 变体。因此，ChR2 激活经常受到经过纤维的虚假激活的污染，从而损害了功能效应的准确解释。在这里，我们探索了专门定位于突触前轴突末端的 ChR2 变体的工程。代谢型谷氨酸受体 2 (mGluR2) C 末端结构域与蛋白水解基序和轴突靶向信号（mGluR2-PA 标签）融合，将 ChR2-YFP 定位在轴突末端，而不干扰正常传输。 mGluR2-PA 标记的 ChR2 在远端投影区域诱发递质释放，从而实现较低水平的光刺激。通过尖峰碰撞测试进行的体内电路连接图显示，mGluR2-PA 标记的 ChR2 可用于识别轴突投影，并显着减少多突触过量噪声。这些结果表明 mGluR2-PA 标签有助于驱动向轴突末端的运输，从而为光遗传学实验提供丰富的可能性。滨田等人。设计并利用定位于突触前轴突末端的通道视紫红质-2 变体。他们展示了其在体内电路映射中的用途，从而为未来的光遗传学实验提供了有用的工具
Optogenetic tools such as channelrhodopsin-2 (ChR2) enable the manipulation and mapping of neural circuits. However, ChR2 variants selectively transported down a neuron’s long-range axonal projections for precise presynaptic activation remain lacking. As a result, ChR2 activation is often contaminated by the spurious activation of en passant fibers that compromise the accurate interpretation of functional effects. Here, we explored the engineering of a ChR2 variant specifically localized to presynaptic axon terminals. The metabotropic glutamate receptor 2 (mGluR2) C-terminal domain fused with a proteolytic motif and axon-targeting signal (mGluR2-PA tag) localized ChR2-YFP at axon terminals without disturbing normal transmission. mGluR2-PA-tagged ChR2 evoked transmitter release in distal projection areas enabling lower levels of photostimulation. Circuit connectivity mapping in vivo with the Spike Collision Test revealed that mGluR2-PA-tagged ChR2 is useful for identifying axonal projection with significant reduction in the polysynaptic excess noise. These results suggest that the mGluR2-PA tag helps actuate trafficking to the axon terminal, thereby providing abundant possibilities for optogenetic experiments. Hamada et al. engineer and utilise a channelrhodopsin-2 variant that is localized to presynaptic axon terminals. They demonstrate its use for circuitry mapping in vivo and thus provide a useful tool for future optogenetic experiments
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