Nov 17, 2025

Amphetamine-Induced Rotational Behavior Assay in Mice

DOI
https://dx.doi.org/10.17504/protocols.io.yxmvmby65g3p/v1
  • 1Karolinska Institute Stockholm
  • SOX6 mDA differentiation
Roberto Garcia Swinburn
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DOI: https://dx.doi.org/10.17504/protocols.io.yxmvmby65g3p/v1
Protocol Citation: Roberto Garcia Swinburn, Ernest Arenas 2025. Amphetamine-Induced Rotational Behavior Assay in Mice. protocols.io https://dx.doi.org/10.17504/protocols.io.yxmvmby65g3p/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: September 10, 2025
Last Modified: November 17, 2025
Protocol  Integer ID: 226860
Keywords: hemiparkinsonism on mice, hemiparkinsonism, mice, mice this protocol, induced rotational behavior
Abstract
This protocol was used to assess hemiparkinsonism on mice for further grafting.
Protocol materials
D-amphetamine hemisulphateCayman Chemical CompanyCatalog #26204
Drug Preparation
Prepare fresh d-amphetamine solution 0.5 mg/mL D-amphetamine hemisulphateCayman Chemical CompanyCatalog #26204 in sterile saline at a concentration appropriate for a 5 mg/kg dose based on mouse body weight. Keep On ice until use.

Intraperitoneal Injection
Inject each mouse intraperitoneally (IP) with 5 mg/kg d-amphetamine solution.
Use standard sterile technique and ensure accurate dosing.
Behavioral Recording
40m
Allow the mouse to rest for 10 minutes post-injection in its home cage or a holding cage.
10m
After the 10-minute wait, place the mouse into a 15 cm-wide cylindrical arena, under dark or dim-light conditions and quiet environment to minimize stimuli.
Track and analyze behavior using EthoVision XT 17 for 30 minutes:
  • Define active movement as body center velocity >0.5 cm/s.
  • Define the pipeline to capture number of clockwise and counterclockwise rotation, and time of active movement. Using active movement reduces the chance of false negatives due to lack of movement.
30m
Data Analysis
Calculate net ipsilateral rotations per minute by substracting contralateral from ipsilateral rotational count and dividing per time of active movement.
Record each mouse's ID, weight, injection time, and behavioral results in a logbook or digital database for longitudinal tracking.