Apr 28, 2026
Accelerating Rotarod: Motor Coordination and Balance Assessment in Mice
- Sarah van Veen1,2,
- Zsuzsanna Callaerts-Végh1
- 1KU Leuven;
- 2Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, Chevy Chase, MD 20815
Protocol Citation: Sarah van Veen, Zsuzsanna Callaerts-Végh 2026. Accelerating Rotarod: Motor Coordination and Balance Assessment in Mice. protocols.io https://dx.doi.org/10.17504/protocols.io.261geybjyv47/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: April 21, 2026
Last Modified: April 28, 2026
Protocol Integer ID: 315430
Keywords: ASAPCRN, balance assessment in mice, assessment of motor coordination, motor coordination, rotarod in adult mice, accelerating rotarod, adult mice, balance assessment, mice this protocol, mice, rotating rod, balance, accelerating rod
Funders Acknowledgements:
Aligning Science Across Parkinson's
Grant ID: ASAP-000458
Fonds Wetenschappelijk Onderzoek (FWO)
Grant ID: G011424N
Abstract
This protocol describes the assessment of motor coordination and balance using an accelerating rotarod in adult mice. After a training phase on the rotating rod at constant speed, mice undergo four test trials on an accelerating rod, and latency to fall is recorded.
Guidelines
• Ensure consistent placement on the rod — mouse should be placed mid-rod with all four paws gripping.
• Clean rod with 70% ethanol between animals to remove olfactory cues.
• All testing should be performed at the same time of day.
• Exclude animals that jump off within the first 5 s of a trial; repeat that trial.
Materials
| A | B | |
| Subjects | Adult WT and KO littermates (~2 months old) | |
| Apparatus | Rotarod (Ugo Basile, Italy) | |
| Training speed | 5 rpm (constant) × 2 min on rotating rod | |
| Test protocol | Accelerating: 5 → 40 rpm over 5 min (300 s) | |
| Number of trials | 4 test trials per animal | |
| Inter-trial interval | 10 min | |
| Endpoint | Latency to fall (max 300 s); mean of 4 trials |
Before start
• Calibrate rotarod: confirm acceleration rate (5→40 rpm over 5 min) and constant-speed setting (5 rpm).
• Allow mice to acclimate to the procedure room for ≥30 min before testing.
• Confirm the automatic timer stops at 300 s maximum.
Training phase
Place each mouse on the rotating rod already set to constant speed (5 rpm). Allow the mouse to
walk on the rod for 2 min. If the mouse drops during training, place it back on the rod (up to 3 times).
This familiarizes the animal with the apparatus. Return mouse to home cage after training.
Test trials - setup
Place all mice onto the rotating rod at 5 rpm, one at a time, ensuring all four paws are gripping
mid-rod. Once all mice are positioned, start the acceleration protocol. The rod accelerates from 5
rpm to 40 rpm over 5 min (300 s). Record latency to drop (in seconds) for each mouse. Maximum
trial duration is 300 s.
Note
If a mouse clings to the rod and does not walk on it, observe for a maximum of 3 full rotations to allow it to resume walking. If the mouse continues clinging after 3 rotations, remove it from the apparatus and record the time at that point as the latency.
Inter-trial rest (10 min)
Return each mouse to its home cage for a 10-min inter-trial interval. Repeat Steps 2 and 3 for trials
2, 3, and 4.
Complete all 4 trials
Each mouse completes 4 test trials with 10-min inter-trial intervals. Record latency to fall for each
trial. Calculate the mean latency across all 4 trials per animal.
Data analysis
Express performance as mean latency to fall ± SEM across the 4 test trials per animal. Compare WT vs KO using appropriate statistics.