Neuron
Volume 71, Issue 4, 25 August 2011, Pages 617-631
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New Rabies Virus Variants for Monitoring and Manipulating Activity and Gene Expression in Defined Neural Circuits

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Summary

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 12 novel ΔG rabies variants from plasmid DNA. These new rabies viruses express useful neuroscience tools, including the Ca2+ indicator GCaMP3 for monitoring activity; Channelrhodopsin-2 for photoactivation; allatostatin receptor for inactivation by ligand application; and rtTA, ERT2CreERT2, or FLPo, for control of gene expression. These new tools allow neurons targeted on the basis of 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.

Highlights

► New methods allowed efficient production of ΔG rabies viruses from plasmid DNA ► Twelve new rabies variants express useful genetically-encoded neuroscience tools ► Encoded genes monitor or control activity or gene expression in defined circuits ► These reagents facilitate studies directly linking circuit structure to function

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2

These authors contributed equally to this work

3

Present address: Department of Information Physiology, National Institute for Physiological Sciences, Okazaki, Aichi 444-8787, Japan