Monosynaptic Rabies Tracing
Alexandra Nelson, Michael Ryan
Abstract
This protocol describes the steps for performing stereotaxic surgery in mice. It is applicable to intracranial injections (e.g. virus, drug) and placement of implants (e.g. optical fibers, electrode arrays) into targeted regions of mouse brains.
Steps
Rabies injections were performed in a BSL-2 surgical suite following the protocol described in dx.doi.org/10.17504/protocols.io.n2bvj6qynlk5/v1
For modified, G-deleted rabies viruses, first inject a Cre-dependent helper virus (AAV-DIO-sTpEpB-GFP) to restrict expression of the EnvA receptor (TVA) and rabies glycoprotein necessary for subsequent rabies virus infection.
2 weeks later, inject the modified, G-deleted rabies virus (EnvA-G-deleted-rabies-mCherry) into the same coordinates as the helper virus.
For a detailed protocol regarding subsequent perfusion, sectioning, staining, imaging, and analysis with MBF NeuroInfo software, see Eastwood, et al., 2019 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6570587/
