Inducing Penetrating Traumatic Brain Injury in Adult Drosophila Flies

Take newly emerged, genetically modified adult Drosophila flies.

Mushroom bodies in the fly's brain contain neurons that express green fluorescent proteins.

Mushroom bodies are structures containing complex neuronal networks essential for learning and memory.

Anesthetize the flies on a carbon dioxide pad.

Under a stereo microscope equipped with the required filters, visualize the mushroom bodies expressing green fluorescence.

Take a sanitized Minutien pin and push it through the head capsule into the mushroom body.

This causes damage to mushroom body neurons and disrupts neuronal networks, inflicting penetrating traumatic brain injury.

Next, gently remove the pin from the fly.

Place the injured flies in a vial with food and allow them to recover for further studies.

After anesthetizing the flies on a carbon dioxide pad, sort newly eclosed F1 young male flies within six hours of eclosion, and place them in clean vials containing food with 40 or fewer flies per vial. Next, sanitize minutien pins by placing approximately 100 pins in a 1.5-milliliter microcentrifuge tube filled with 70% ethanol for five minutes. Then, sanitize the carbon dioxide pad in paintbrush by spraying 70% ethano