Rats were trained on the accelerating rotarod (IITC Life Science ,Woodland Hills, CA) for 3 days starting at PD90 with 3 trials of 180 seconds each day, as previously described (Dulman et al., 2019 (link)). Briefly, the rotarod cylinder apparatus is 9.5 cm in diameter and 15 cm wide; the rotarod starts spinning at 5 rotations per minute and accelerates to 20 rotations per minute over the course of the full 180-second trial. Latency to fall is recorded for every trial and is the primary outcome measure. Time spent on the rod is calculated as an average latency to fall across the three trials measured within each rotarod test session. Rats were tested at the same time every day at 10AM, with the exception of the acute withdrawal 8-hour timepoint when rats were tested at 6PM. Schematic diagram of periodic rotarod testing and chronic diet procedure is shown in Figure 1. During training, rats were placed back on the rotarod apparatus if they fell off before the full trial was completed. By the end of training, all rats were able to remain on the rod for the entire 180 second rotarod session in nearly every trial. Once the rats were trained, we controlled for motor behavior “overtraining” (Luong et al., 2011 ; Scholz et al., 2015 (link); Tung et al., 2014 ) by only conducting rotarod testing every three days throughout the chronic diet procedure.