Mar 07, 2024

Public workspaceMonosynaptic Rabies Tracing

 Forked from Mouse Stereotaxic Surgery
DOI
dx.doi.org/10.17504/protocols.io.3byl4qp9jvo5/v1
  • Alexandra Nelson1,
  • Michael Ryan1
  • 1University of California San Francisco
Michael Ryan
Open access
DOI: dx.doi.org/10.17504/protocols.io.3byl4qp9jvo5/v1
Protocol Citation: Alexandra Nelson, Michael Ryan 2024. Monosynaptic Rabies Tracing. protocols.io https://dx.doi.org/10.17504/protocols.io.3byl4qp9jvo5/v1
Manuscript citation:
Jonathan S Schor, Isabelle Gonzalez Montalvo, Perry WE Spratt, Rea J Brakaj, Jasmine A Stansil, Emily L Twedell, Kevin J Bender, Alexandra B Nelson (2022) Therapeutic deep brain stimulation disrupts movement-related subthalamic nucleus activity in Parkinsonian mice eLife 11:e75253

https://doi.org/10.7554/eLife.75253
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 27, 2024
Last Modified: March 07, 2024
Protocol Integer ID: 95789
Keywords: Mouse, Surgery, Stereotaxic Surgery, Implants, ASAPCRN
Funders Acknowledgement:
NINDS
Grant ID: R01NS101354
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.
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/