McSweeney Colleen, Mao Yingwei
Department of Biology, The Pennsylvania State University.
Department of Biology, The Pennsylvania State University;
J Vis Exp. 2015 May 10(99):e52653. doi: 10.3791/52653.
Stereotactic injection is a useful technique to deliver high titer lentiviruses to targeted brain areas in mice. Lentiviruses can either overexpress or knockdown gene expression in a relatively focused region without significant damage to the brain tissue. After recovery, the injected mouse can be tested on various behavioral tasks such as the Open Field Test (OFT) and the Forced Swim Test (FST). The OFT is designed to assess locomotion and the anxious phenotype in mice by measuring the amount of time that a mouse spends in the center of a novel open field. A more anxious mouse will spend significantly less time in the center of the novel field compared to controls. The FST assesses the anti-depressive phenotype by quantifying the amount of time that mice spend immobile when placed into a bucket of water. A mouse with an anti-depressive phenotype will spend significantly less time immobile compared to control animals. The goal of this protocol is to use the stereotactic injection of a lentivirus in conjunction with behavioral tests to assess how genetic factors modulate animal behaviors.
立体定位注射是一种将高滴度慢病毒递送至小鼠大脑靶向区域的有用技术。慢病毒可以在相对局限的区域内过表达或敲低基因表达,而对脑组织造成的损伤较小。恢复后,可以对注射后的小鼠进行各种行为测试,如旷场试验(OFT)和强迫游泳试验(FST)。旷场试验旨在通过测量小鼠在新的旷场中央停留的时间来评估其运动能力和焦虑表型。与对照组相比,焦虑程度更高的小鼠在新旷场中央停留的时间会显著减少。强迫游泳试验通过量化小鼠放入水桶中静止不动的时间来评估抗抑郁表型。与对照动物相比,具有抗抑郁表型的小鼠静止不动的时间会显著减少。本方案的目的是结合行为测试使用慢病毒的立体定位注射来评估遗传因素如何调节动物行为。