Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow 117997, Russia.
Pirogov Russian National Research Medical University, Moscow 117997, Russia.
Nat Commun. 2017 May 22;8:15362. doi: 10.1038/ncomms15362.
Thermogenetics is a promising innovative neurostimulation technique, which enables robust activation of neurons using thermosensitive transient receptor potential (TRP) cation channels. Broader application of this approach in neuroscience is, however, hindered by a limited variety of suitable ion channels, and by low spatial and temporal resolution of neuronal activation when TRP channels are activated by ambient temperature variations or chemical agonists. Here, we demonstrate rapid, robust and reproducible repeated activation of snake TRPA1 channels heterologously expressed in non-neuronal cells, mouse neurons and zebrafish neurons in vivo by infrared (IR) laser radiation. A fibre-optic probe that integrates a nitrogen-vacancy (NV) diamond quantum sensor with optical and microwave waveguide delivery enables thermometry with single-cell resolution, allowing neurons to be activated by exceptionally mild heating, thus preventing the damaging effects of excessive heat. The neuronal responses to the activation by IR laser radiation are fully characterized using Ca imaging and electrophysiology, providing, for the first time, a complete framework for a thermogenetic manipulation of individual neurons using IR light.
热遗传学是一种很有前途的创新神经刺激技术，它可以使用热敏瞬时受体电位 (TRP) 阳离子通道来实现神经元的强大激活。然而，这种方法在神经科学中的更广泛应用受到可用离子通道种类有限以及当 TRP 通道被环境温度变化或化学激动剂激活时神经元激活的空间和时间分辨率低的限制。在这里，我们通过红外 (IR) 激光辐射证明了在非神经元细胞、小鼠神经元和斑马鱼神经元中异源表达的蛇 TRPA1 通道的快速、强大和可重复的重复激活。一种光纤探头，它将氮空位 (NV) 金刚石量子传感器与光学和微波波导传输集成在一起，能够实现单细胞分辨率的测温，使神经元能够通过异常温和的加热来激活，从而防止过热的破坏性影响。使用钙成像和电生理学对 IR 激光辐射激活的神经元反应进行了全面表征，首次为使用 IR 光对单个神经元进行热遗传操作提供了完整的框架。
