Inducing Ethyl Butyrate (EB) Experience-Dependent Synaptic Pruning of Or42a Receptor

To begin, take the vial containing the drosophila larvae. Using a fine paintbrush, sort 40 to 50 pharate dark pupae in polystyrene drosophila vials containing standard cornmeal molasses food. Place fine stainless steel wire mesh over the end of the drosophila vials to contain the flies while allowing good airflow.

Secure the wire mesh caps with a tape transparent film onto the side of the vials. Then, add one milliliter of 100%mineral oil, or dissolve EB in mineral oil in 1.5 milliliter tubes. Place these tubes upright within a 3, 700 milliliter glass lock container.

Using tape, anchor the tubes in the center of the odorant chambers, along with the drosophila vials. Place the sealed vehicle control and EB odorant chambers containing the pupae vials in a humidified incubator on a 12 hour light and 12 hour dark cycle. After four hours of odorant chamber exposure, rapidly transfer 20 to 25 newly eclosed flies to fresh drosophila vials in chambers with freshly made odorant.

Keep the flies in their sealed odorant chambers in the incubators for a total of 24 hours.