Drosophila Brain Dissection to Analyze Ethyl Butyrate (EB) Experience-Dependent Synaptic Pruning of Or42a Receptor

To begin, take drosophila flies exposed to mineral oil and EB for 24 hours. Label vials as a vehicle control or EB exposed, and maintain these designations throughout brain dissection and processing. Using forceps, transfer a single anesthetized fly into a small dish of freshly made PBS.

Fully submerge the fly in PBS for full brain dissection. Then, position a fly ventral side up. Take two fine sharpened forceps to grasp the upper thorax with one forcep and the head, under the proboscis with the other.

Pull the head and the rest of the body in opposite directions to remove the head. Ensure the head easily detaches from the thorax and leaves the isolated head for dissection. Slide the forceps, previously used to grasp the thorax, under the opposite side of the proboscis.

Gently pull the exoskeleton cuticle in opposite directions to tear up between the eyes and reveal the brain. Continue to remove the exoskeleton cuticle from the head, ensuring all parts are removed. Now, rinse a P20 pipette tip with PBS and 0.2%Triton X-100.

Transfer the brains into the fixing solution in the cap tube immediately after dissection.