Drosophila Courtship Conditioning: A Method to Test Learning and Memory In Flies

-First, isolate single naive, or unmated, Drosophila melanogastor males. Then, to condition the males, add a female that has already mated, and is therefore non-receptive to courtship. The male displays typical courtship behaviors such as orienting towards her, following her around, tapping her with a foreleg, and extending and vibrating his wing to produce the courtship song. He may also attempt licking and mating. No matter the efforts, the female remains non-receptive, and the male experiences a rejection. Thus, he learns not to court a previously mated female.

To test the male's memory, allow him to rest, and then expose him to another mated female. The trained male will not attempt to court this fly because he retains the memory of past rejection. In the example protocol, we will see how flies are handled during the training and testing phases of a courtship conditioning assay for short- and long-term memory assessment.

- Begin by anesthetizing all of the adults from the four-day-old mating files. Separate the sexes, and transfer single premated females into each of the 12 wells of one row of a housing block. Then, using an aspirator and no anesthesia, remove the naive males f