Chemicals and treatment paradigms
Pranay Srivastava
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).
Steps
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
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.
