Harris Julie A, Oh Seung Wook, Zeng Hongkui
Allen Institute for Brain Science, Seattle, Washington, USA.
Curr Protoc Neurosci. 2012 Apr;Chapter 1:Unit 1.20.1-18. doi: 10.1002/0471142301.ns0120s59.
Harnessing the natural ability of viruses to infect post-mitotic cells such as neurons has provided an explosion of new methods to manipulate and reconstruct neural circuits in vivo. Here we describe the use of recombinant adeno-associated viral vectors (rAAV) for axonal tract tracing in nontransgenic and transgenic Cre driver mice. Two protocols are presented for stereotactic-guided placement of rAAV vectors into the live mouse brain using iontophoretic or nanoliter pressure injections. The methods discussed here will result in expression of fluorescent proteins in cell bodies, dendrites, and axons in targeted neurons, and can be easily adapted to address various experimental questions.
利用病毒感染有丝分裂后细胞(如神经元)的天然能力,催生了大量用于在体内操纵和重建神经回路的新方法。在此,我们描述了重组腺相关病毒载体(rAAV)在非转基因和转基因Cre驱动小鼠中用于轴突束追踪的应用。介绍了两种通过离子电渗法或纳升压力注射将rAAV载体立体定位引导至活小鼠脑内的方案。本文讨论的方法将使荧光蛋白在目标神经元的细胞体、树突和轴突中表达,并且可以轻松调整以解决各种实验问题。