Traumatic Brain Injury Induction in Drosophila

The TBI methods used for Drosophila have been previously detailed [10 (link)]. Briefly, flies from different genders, ages, or treatment regimens were anesthetized and placed in 2 mL screw cap tubes (10 flies/tube) [9 (link)]. Control and traumatized flies were allowed to recover from anesthesia before being placed in the Omni Bead Ruptor-24 homogenizer (Omni International, Kennesaw, GA, USA) and subjected to specific pre-programmed shaking/trauma conditions involving 5 s trauma bouts [9 (link)]. Following the completion of the injury, all fly cohorts are returned to vials containing control or treated media, allowed to recover, and maintained using standard care and culturing conditions [9 (link)]. For this study, fly cohorts were exposed to 10 mild bouts of trauma (mTBI; 10×, 2.1 m/s) or one severe trauma bout (mTBI; 1×, 4.3 m/s) [9 (link)].