Dec 01, 2023
Chemicals and treatment paradigms
- 1Massachusetts General Hospital, ASAP
Protocol Citation: Pranay Srivastava 2023. Chemicals and treatment paradigms. protocols.io https://dx.doi.org/10.17504/protocols.io.3byl4q1kzvo5/v1
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: November 30, 2023
Last Modified: May 31, 2024
Protocol Integer ID: 91665
Keywords: ASAPCRN
Abstract
This protocol is to enhance microglia response and nigral dopaminergic cell death in an acute MPTP model following peripheral inflammation resulting from single i.p. injection of LPS at 2 mg/kg (García-Domínguez et al. 2018).
Mice were randomly divided into MPTP+LPS+NDP-MSH, MPTP+LPS, and control groups to receive i.p. once daily MPTP.HCl (Millipore Sigma, Cat# M0896; 20 mg/kg) or saline and LPS (Millipore Sigma, Cat# L4391; 1 mg/kg) or PBS from day 1 to day 4.
NDP-MSH (Genscript, Cat# RP10658; 400 µg/kg) or PBS was injected from day 1 to day 12.
Mice were tested for behavioral activities and were sacrificed thereafter on day 12.
To study the role of Tregs
To study the role of Tregs
Animals were treated with anti-mouse CD25 monoclonal antibody (clone PC61, Biolegend, Cat# 102059; 400 µg/mouse) or isotype control (Biolegend, Cat# 401916; 100µg/mice) for 3 alternate days, 1 week before the start of experiment.
Mice were subsequently treated with MPTP, LPS and NDP-MSH as described above.
Another dose of anti-mouse CD25 monoclonal antibody or isotype control was administered 2 days before the sacrifice.