一种玻璃涂层的钨微电极,用于在灵长类动物深部脑结构中进行光遗传探索,该电极包裹着光纤。
A glass-coated tungsten microelectrode enclosing optical fibers for optogenetic exploration in primate deep brain structures.
机构信息
Department of Physiology, The University of Tokyo School of Medicine, Japan.
出版信息
J Neurosci Methods. 2012 Oct 15;211(1):49-57. doi: 10.1016/j.jneumeth.2012.08.004. Epub 2012 Aug 14.
The optogenetic approach to primate brain circuitry has unparalleled potential for uncovering genetically and temporally resolved neuronal mechanisms of higher brain functions. In order to optogenetically investigate the large and complex primate brain, an optical-/electrical probe, or "optrode", must be inserted deeply, which requires the optrode to be not only long and stiff, but also sharp and smooth to reduce possible tissue damage. This study presents a tungsten microelectrode-based optrode that encloses optical fibers within its insulation glass. Optical fibers and a tungsten wire were tightly bound to each other and integrally coated with a smooth, thin layer of glass. This design satisfied the structural requirements for use in deep brain structures. The performance of the optrode was then examined in the thalamus of the rat and macaque monkeys which were injected with lentiviral vectors carrying the channelrhodopsin-2-enhanced yellow fluorescent protein (ChR2-EYFP) transgene. With fluorescence measurements via the optical fiber, ChR2-EYFP expression was detected clearly in vivo, which was confirmed by histological analysis in the rat. With photostimulation and extracellular recording, photo-responsive single-unit activities were isolated in the monkeys. The depth distribution of these units and the peak of the EYFP fluorescence profile overlapped consistently with each other. Thus, by developing a new probe, optogenetic methodology was successfully applied to a primate subcortical structure. This smooth glass-coated optrode is a promising tool for chronic in vivo experiments with various research targets including deep brain structures in behaving monkeys.
光遗传学方法在揭示高等大脑功能的基因和时间分辨神经元机制方面具有无与伦比的潜力。为了对大型和复杂的灵长类大脑进行光遗传学研究,必须将光学/电气探针(“光探头”)插入到深部,这要求光探头不仅要长而硬,还要锋利和光滑,以减少可能的组织损伤。本研究提出了一种基于钨微电极的光探头,其绝缘玻璃内封装有光纤。光纤和钨丝被紧紧地绑定在一起,并整体涂覆有光滑、薄的玻璃层。这种设计满足了在深部脑结构中使用的结构要求。然后,在大鼠和猕猴的丘脑内检查了光探头的性能,这些动物被注射了携带通道视紫红质-2 增强型黄色荧光蛋白(ChR2-EYFP)转基因的慢病毒载体。通过光纤进行荧光测量,在体内清楚地检测到 ChR2-EYFP 的表达,这在大鼠的组织学分析中得到了证实。通过光刺激和细胞外记录,在猴子中分离出对光有反应的单个单元活动。这些单元的深度分布与 EYFP 荧光分布的峰值一致。因此,通过开发一种新的探针,成功地将光遗传学方法应用于灵长类亚皮质结构。这种光滑玻璃涂层的光探头是一种有前途的工具,可用于对包括行为猴子深部脑结构在内的各种研究目标进行慢性体内实验。