The Accelerating Rotating Rod Assay or Rotarod Test: A Method to Test Motor Coordination and Learning in Mice

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Accelerating Rotarod

1. Acclimate subjects to the assay room 10 min prior to the start of the trial. During this acclimation period, prepare the rotarod apparatus, cleaning each lane of the apparatus with 30% ethanol.
2. Open the rotarod software and set start settings to 4 rpm, acceleration settings to 5 min, and final speed settings to 40 rpm. These settings provide a constant acceleration from 4–40 rpm over a period of 5 min. Mice can remain on the rotarod at 40 rpm for an additional 5 min beyond the acceleration period for a maximum run length of 10 min.
3. Post-acclimation, place subjects onto the rotarod in their respective lanes while the rod is rotating at 4 rpm. Once all subjects are in their respective lanes, begin the first trial. Record the latency to fall using the instrument software; a secondary timer can be employed as a backup.
4. Following trial completion, allow each subject to rest in their respective lane reservoirs for ten minutes prior to the next trial to prevent fatigue.
5. Repeat steps 1.3 and 1.4 for a total of four trials.
6. Repeat steps 1.3–1.5 for a total of four consecutive days, continuing treatment throughout the trial period. Upon completion, quantify latency to fall off the rotating rod for each trial per subject.