New rabies virus variants for monitoring and manipulating activity and gene expression in defined neural circuits.
New rabies virus variants for monitoring and manipulating activity and gene expression in defined neural circuits.
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DOI：
10.1016/j.neuron.2011.07.005
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发表时间：
2011-08-25
期刊：
影响因子：
16.2
通讯作者：
Callaway EM
中科院分区：
文献类型：
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作者：
Osakada F;Mori T;Cetin AH;Marshel JH;Virgen B;Callaway EM
糖蛋白缺失型狂犬病病毒是研究神经回路结构的有力工具。在这里，我们描述的发展，并证明新的资源，允许实验直接调查神经回路的结构和功能之间的关系的效用。新的方法和试剂允许从质粒DNA高效生产12种新的ΔG狂犬病变体。这些新的狂犬病病毒表达有用的神经科学工具，包括：Ca++指示剂GCaMP 3，用于监测活动;视紫红质-2，用于光活化; allatostatin受体，用于通过配体应用灭活; rtTA，ERT 2CreERT 2或FLPo，用于控制基因表达。这些新工具允许基于其连接性靶向神经元，以测定其功能或操纵其活性或基因表达。将这些工具与体内成像和光遗传学方法和/或转基因小鼠中的诱导型基因表达相结合，将有助于研究神经回路发育、可塑性和功能的实验，而这些实验是现有试剂无法实现的。
Glycoprotein-deleted (ΔG) rabies virus is a powerful tool for studies of neural circuit structure. Here we describe the development and demonstrate the utility of new resources that allow experiments directly investigating relationships between the structure and function of neural circuits. New methods and reagents allowed efficient production of twelve novel ΔG rabies variants from plasmid DNA. These new rabies viruses express useful neuroscience tools, including: the Ca++ indicator GCaMP3, for monitoring activity; Channelrhodopsin-2, for photoactivation; allatostatin receptor, for inactivation by ligand application; rtTA, ERT2CreERT2, or FLPo, for control of gene expression. These new tools allow neurons targeted based on their connectivity, to have their function assayed or their activity or gene expression manipulated. Combining these tools with in vivo imaging and optogenetic methods, and/or inducible gene expression in transgenic mice, will facilitate experiments investigating neural circuit development, plasticity, and function that have not been possible with existing reagents.
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