Axonal and subcellular labelling using modified rabies viral vectors.

Axonal and subcellular labelling using modified rabies viral vectors.
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DOI:
10.1038/ncomms3332
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发表时间:
2013
影响因子:
16.6
通讯作者:
Seung HS
Seung HS
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Wickersham IR;Sullivan HA;Seung HS

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任何神经回路的一个重要方面是其输出突触的位置,从宏观到亚细胞的水平。定位和操纵突触的许多新的分子工具受到病毒载体的限制。腺相关病毒是目前标记和操纵轴突和突触的最佳手段,但它们从未表达过一种以上的转基因,其高度足以标记精细的轴突结构,同时也标记或干扰突触。它们的缓慢表达也使它们与逆行和跨突触载体不相容,阻止了强大的组合实验。在这里,我们表明,删除突变狂犬病病毒可以特异性地针对细胞局部注射部位,明亮的标记轴突,即使当共表达两个其他转基因。我们展示了几种新的能力:同时标记轴突和突触前末梢,标记树突和突触后密度,并同时标记一个区域的输入和输出使用共注入向量。
An important aspect of any neural circuit is the placement of its output synapses, at levels ranging from macroscopic to subcellular. The many new molecular tools for locating and manipulating synapses are limited by the viral vectors available for delivering them. Adeno-associated viruses are the best current means of labeling and manipulating axons and synapses, but they have never expressed more than one transgene highly enough to label fine axonal structure while also labeling or perturbing synapses. Their slow expression also makes them incompatible with retrograde and transsynaptic vectors, preventing powerful combinatorial experiments. Here we show that deletion-mutant rabies virus can be specifically targeted to cells local to an injection site, brightly labeling axons even when coexpressing two other transgenes. We demonstrate several novel capabilities: simultaneously labeling axons and presynaptic terminals, labeling both dendrites and postsynaptic densities, and simultaneously labeling a region’s inputs and outputs using coinjected vectors.
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