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: January 14, 2025
Last Modified: February 16, 2025
Protocol Integer ID: 118324
Funders Acknowledgements:
Aligning Science Across Parkinson’s
Grant ID: ASAP-020495
Abstract
Mice Housing and Experimentation Conditions
Animal Housing
Animal Housing
1w 3d
1w 3d
WT C57BL/6J mice were obtained from JAX (RRID:IMSR_JAX:000664, Jackson Laboratory,Bar Harbor, ME).
The mice are habituated for a minimum of 10 days.
1w 3d
All animals are housed in a reverse light cycle, and the room they are housed in is both temperature and humidity controlled, as is all the rooms the mice are tested in.
Care of these animals was in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals
Experimental Conditions
Experimental Conditions
When the experiment started, all mice were aged 8 weeks, and all mice used in this study were male
For each experimental condition, two cohorts of animals were used, and each cohort has three replicates of mice with the same experimental condition
Initially, these mice had normal feed for the acclimation period, and for two weeks post retro orbital injection.
Two weeks after retro-orbital injection, mice were switched to a doxycycline feed (Cat. No S3888, Bio-Serv,Flemington, Nj) in order to activate the virus
The doxycycline feed was also supplemented by an intraperitoneal injection of 2 mg/kg doxycycline (Cat No. D9891-10G, Millipore Sigma,Darmstadt, Germany) twice weekly.
Mice from each experimental condition were randomized and sacrificed at predetermined time points to collect tissue for histology and transcriptomics.
Both data collection and analyses were not performed blind to the experimental conditions.