Cocas Laura, Fernandez Gloria
Department of Neurology, University of California, San Francisco, 675 Nelson Rising Lane, San Francisco, CA, 94158, USA.
Department of Biological Sciences, California State University, 5151, Los Angeles, CA, 90032, USA.
Methods Mol Biol. 2017;1538:353-366. doi: 10.1007/978-1-4939-6688-2_23.
An attenuated rabies virus that expresses fluorescent protein has made it possible to analyze retrograde (presynaptic) monosynaptic connections in vivo. By combining attenuated rabies virus with a Cre-loxP based system to target cells in a subtype-specific fashion, it is possible to examine neuronal input in vivo onto any class of neuron, in development and in the mature brain. We describe here the methods to amplify deletion mutant, pseudotyped rabies virus, selectively target cells of interest using genetic and viral approaches, as well as the stereotaxic procedures required to target neuronal subtypes of interest in vivo.
一种表达荧光蛋白的减毒狂犬病病毒使得在体内分析逆行（突触前）单突触连接成为可能。通过将减毒狂犬病病毒与基于Cre-loxP的系统相结合，以亚型特异性方式靶向细胞，可以在发育中和成熟大脑中体内检查任何一类神经元上的神经元输入。我们在此描述扩增缺失突变型、假型狂犬病病毒的方法，使用遗传和病毒方法选择性靶向感兴趣的细胞，以及在体内靶向感兴趣的神经元亚型所需的立体定位程序。
