Max Planck Florida Institute for Neuroscience, Jupiter, Florida, USA.
Department of Anatomy, College of Medicine, Korea University, Seoul, Republic of Korea.
Nat Methods. 2017 May;14(5):495-503. doi: 10.1038/nmeth.4234. Epub 2017 Apr 3.
Few tools exist to visualize and manipulate neurons that are targets of neuromodulators. We present iTango, a light- and ligand-gated gene expression system based on a light-inducible split tobacco etch virus protease. Cells expressing the iTango system exhibit increased expression of a marker gene in the presence of dopamine and blue-light exposure, both in vitro and in vivo. We demonstrated the iTango system in a behaviorally relevant context, by inducing expression of optogenetic tools in neurons under dopaminergic control during a behavior of interest. We thereby gained optogenetic control of these behaviorally relevant neurons. We applied the iTango system to decipher the roles of two classes of dopaminergic neurons in the mouse nucleus accumbens in a sensitized locomotor response to cocaine. Thus, the iTango platform allows for control of neuromodulatory circuits in a genetically and functionally defined manner with spatial and temporal precision.
目前很少有工具可以用于可视化和操作作为神经调质靶点的神经元。我们提出了 iTango，这是一种基于光诱导的分裂烟草蚀纹病毒蛋白酶的光控和配体门控基因表达系统。在体外和体内，表达 iTango 系统的细胞在多巴胺存在和蓝光暴露的情况下，都会增加标记基因的表达。我们在一个行为相关的背景下证明了 iTango 系统的有效性，通过在感兴趣的行为期间，在受多巴胺能控制的神经元中诱导光遗传学工具的表达。由此，我们获得了对这些行为相关神经元的光遗传学控制。我们将 iTango 系统应用于在可卡因敏感的运动反应中，解析小鼠伏隔核中两类多巴胺能神经元的作用。因此，iTango 平台允许以遗传和功能定义的方式，精确地控制神经调质回路。
