Bang-Sensitive Assay for the Study of Seizure-Like Behavior in Drosophila

To begin, collect the virgin flies of the tub-Gal4 hybrid line and the male flies of the UAS-cac-RNAi line. Transfer the virgin and male flies into the same vial to harvest the offspring. After three to five days of occlusion, use a brush to collect the offspring and the tub-GL4 UAS-cac-RNA flies.

A day before testing, transfer the flies to new clean vials with food. Next, carefully place four to six carbon dioxide anesthetized flies into individual fresh vials. Place a camera on a tripod in front of a whiteboard.

Manually adjust the camera's focus with an empty vial. Now, vortex the vials with the flies at the highest setting for 10 seconds. Immediately place the vial on the whiteboard and observe the flies for any seizure-like behavior.

Measure the recovery time as the time required for the flies to regain the ability to stand upright. The cac knockdown flies showed significantly higher rates of seizure-like behavior than the wild type flies. The recovery percentage of knockdown flies within one second was significantly lower than the wild type flies.