Aggression Assay in Drosophila to Analyze Autism Spectrum Disorder (ASD)-Related Communication and Social Behavior Deficit

To begin, take a standard 24-well plate and fill half of each well with regular fly food. Let the food dry overnight. Make a small, 2.5-millimeter hole on a transparent plastic or acrylic sheet and place it over a well plate.

Using a stereo microscope, separate newly enclosed flies of the desired genotype based on their sex. Insert half the male flies individually into single housing tubes. Keep the other half of the male flies in a group of 10 with the female flies in a regular group housing vial, creating a social condition.

Store all vials at 25 degrees Celsius for five days. Mouth aspirate two male flies from either the single or group housing chambers to the aggression arena plate through the hole in the lid. Move the hole away immediately to ensure that the flies cannot escape.

After one to two minutes of acclimatization, start a timer for 20 minutes. Make sure to video record the flies. Play the video on a computer and count the total number of aggressive bouts for each fly.

The number of aggressive bouts in 20 minutes was significantly reduced in the case of mutant males than in the control males.