Cetin Ali, Komai Shoji, Eliava Marina, Seeburg Peter H, Osten Pavel
Department of Molecular Neurobiology, Max Planck Institute for Medical Research, Jahnstrasse 29, 69120 Heidelberg, Germany.
Nat Protoc. 2006;1(6):3166-73. doi: 10.1038/nprot.2006.450.
Stereotaxic surgery has been an invaluable tool in systems neuroscience, applied in many experiments for the creation of site-targeted lesions, injection of anatomical tracers or implantation of electrodes or microdialysis probes. In this protocol, we describe stereotaxic surgery optimized for gene delivery by recombinant adeno-associated viruses and lentiviruses in mice and rats. This method allows the manipulation of gene expression in the rodent brain with excellent spatiotemporal control; essentially any brain region of choice can be targeted and cells (or a subpopulation of cells) in that region can be stably genetically altered at any postnatal developmental stage up to adulthood. Many aspects of the method, its versatility, ease of application and high reproducibility, make it an attractive approach for studying genetic, cellular and circuit functions in the brain. The entire protocol can be completed in 1-2 hours.
立体定向手术一直是系统神经科学中一项非常宝贵的工具，被应用于许多实验中，用于制造位点靶向损伤、注射解剖示踪剂或植入电极或微透析探针。在本方案中，我们描述了针对重组腺相关病毒和慢病毒在小鼠和大鼠中进行基因递送而优化的立体定向手术。该方法能够在啮齿动物大脑中以出色的时空控制来操纵基因表达；基本上可以靶向任何选定的脑区，并且在成年前的任何出生后发育阶段，该区域的细胞（或细胞亚群）都可以被稳定地进行基因改造。该方法的诸多方面，包括其多功能性、易于应用和高重现性，使其成为研究大脑中基因、细胞和神经回路功能的一种有吸引力的方法。整个方案可在1 - 2小时内完成。
