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Neuroscience

Optogenetically有针对性的前脑电路的激光扫描光刺激

Published: December 27, 2013 doi: 10.3791/50915
Automatic Translation
1, 2, 1, 2
1Department of Comparative Biomedical Sciences, School of Veterinary Medicine, Louisiana State University, 2Department of Neurobiology, University of Chicago

Abstract

感觉前脑是由错综复杂的连接类型的细胞，其功能特性尚未完全阐明的。了解这些脑电路的相互作用最近已资助由光遗传学方法对神经元活动的光介导调制的发展。在这里，我们描述了一个协议，用于检查脑电路的体外功能组织使用channelrhodopsin的激光扫描光刺激，通过病毒介导的转染表达optogenetically。这种方法还利用了CRE-LOX重组的转基因小鼠的效用为目标表达的特定的神经元细胞类型。转染后，神经元采用全细胞膜片钳来衡量自己的诱发反应channelrhodopsin表达光纤激光扫描光刺激生理记录片的准备工作。这种方法使功能性地形和synapti的评估C性状。形态特征相关性可以通过使用成像活切片或后固定的组织的共聚焦显微镜channelrhodopsin表达纤维的神经解剖式求出。这些方法使脑电路，扩展后能更传统的方法的功能研究。

Materials

Name Company Catalog Number Comments
Polybrene Santa Cruz Biotech SC134220
Paraformaldehyde 32% solution EMS 15714
VECTASHIELD Vector Labs H-1400
AAV viral constructs UNC Vector Core NA
Mouse stereotactic headholder Stoelting 51730
Nanocool injector Harvard Apparatus 703040
UV laser: 355 nm, Nd:YVO4; 1 W; DPSS Lasers, Inc 3500
Galvonometers Cambridge Scanning 6210
mirrors Cambridge Scanning 6m2003S-A
mount Cambridge Scanning 6102103L
cable Cambridge Scanning 6010-19-120
driver/controller Cambridge Scanning 67121-1
Shutter - ZM coated (UV-compatible) Uniblitz L23-ZM2
Shutter Controller Uniblitz VMM-D1
Current Amplifier Stanford Research Systems SR570
6713 board National Instruments 777741-01
2110 break-out box National Instruments 777643-01
PCI-MIO-16E-4 National Instruments 777383-01
BNC-2090 National Instruments 777270-01
Cables National Instruments 184749-01
MATLAB Mathworks
TCS SP2 Leica Microsystems

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Optogenetically有针对性的前脑电路的激光扫描光刺激
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Cite this Article

Lee, C. C., Lam, Y. W., Imaizumi,… More

Lee, C. C., Lam, Y. W., Imaizumi, K., Sherman, S. M. Laser-scanning Photostimulation of Optogenetically Targeted Forebrain Circuits. J. Vis. Exp. (82), e50915, doi:10.3791/50915 (2013).

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