Simultaneous recording of fluorescence and electrical signals by photometric patch electrode in deep brain regions in vivo
Simultaneous recording of fluorescence and electrical signals by photometric patch electrode in deep brain regions in vivo
复制标题
通过光度贴片电极同时记录体内深部脑区域的荧光和电信号
DOI：
10.1152/jn.00005.2015
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发表时间：
2015
影响因子：
2.5
通讯作者：
and Ohmori H
中科院分区：
文献类型：
--
作者：
Hirai Y;Nishino E;and Ohmori H
尽管多光子显微镜的高分辨率成像被广泛使用，但由于光的穿透有限，在体内记录神经元信号的高分辨率成像仅限于脑组织表面。此外，大多数成像研究并没有同时记录神经电活动，而这对理解大脑功能至关重要。因此，我们开发了一种光度贴片电极（PME），以克服光学测量的深度限制，并能够同时记录大脑深部区域的神经电反应。PME记录系统使用贴片电极激发荧光染料，测量荧光信号作为光导，记录电信号，并在记录的细胞局部应用化学物质。根据响应的动力学，光信号由高光灵敏度的光谱仪或光电倍增管进行分析。我们使用PME在俄勒冈绿BAPTA-1 am加载鸟听觉核体内监测钙信号和电反应。我们在上升听觉通路的三个不同核中展示了不同的反应模式。在声刺激下，听觉皮层（场L）神经元中出现了强大的钙荧光反应，其持续时间超过了电反应。在听觉中脑（下丘），这两种反应都是短暂的。在脑干耳蜗大细胞核中，钙反应似乎被代谢性谷氨酸受体的活性有效地抑制。总之，PME提供了一个强大的工具，可以在常规成像设备无法达到的组织深度研究活体脑功能。
Despite its widespread use, high-resolution imaging with multiphoton microscopy to record neuronal signals in vivo is limited to the surface of brain tissue because of limited light penetration. Moreover, most imaging studies do not simultaneously record electrical neural activity, which is, however, crucial to understanding brain function. Accordingly, we developed a photometric patch electrode (PME) to overcome the depth limitation of optical measurements and also enable the simultaneous recording of neural electrical responses in deep brain regions. The PME recoding system uses a patch electrode to excite a fluorescent dye and to measure the fluorescence signal as a light guide, to record electrical signal, and to apply chemicals to the recorded cells locally. The optical signal was analyzed by either a spectrometer of high light sensitivity or a photomultiplier tube depending on the kinetics of the responses. We used the PME in Oregon Green BAPTA-1 AM-loaded avian auditory nuclei in vivo to monitor calcium signals and electrical responses. We demonstrated distinct response patterns in three different nuclei of the ascending auditory pathway. On acoustic stimulation, a robust calcium fluorescence response occurred in auditory cortex (field L) neurons that outlasted the electrical response. In the auditory midbrain (inferior colliculus), both responses were transient. In the brain-stem cochlear nucleus magnocellularis, calcium response seemed to be effectively suppressed by the activity of metabotropic glutamate receptors. In conclusion, the PME provides a powerful tool to study brain function in vivo at a tissue depth inaccessible to conventional imaging devices.