Enhanced olfactory memory detection in trap-design Y-mazes allows the study of imperceptible memory traces in...

The neural basis of behavior is identified by systematically manipulating the activity of specific neurons and screening for loss or gain of phenotype. Therefore, robust, high-scoring behavioral assays are necessary for determining the neural circuits of novel behaviors. We report a simple Y-maze design for Drosophila olfactory learning and memory assay. Memory scores in our Y-mazes are considerably better and longer-lasting than scores obtained with commonly used T-mazes. Our results suggest that trapping flies to an odor choice in a Y-maze could improve scores. We postulated that the improved scores could reveal previously undetectable memory traces, enabling the study of underlying neural mechanisms. Indeed, we identified unreported protein synthesis-dependent long-term memories (LTMs), reinforced by ingestion of (1) an aversive compound and (2) a sweet but nonnutritious sugar, both 24 h after training. We also used Y-mazes to probe how using a greater reward may change memory dynamics. Our findings predict that a greater sugar reward may extend existing memory traces or reinforce additional novel ones.