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

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.

Tripeptides derived from reactive centre loop of potato type II protease inhibitors preferentially inhibit midgut proteases of Helicoverpa armigera.

Potato type II protease inhibitors (Pin-II PIs) impede the growth of lepidopteran insects by inhibiting serine protease-like enzymes in the larval gut. The three amino acid reactive centre loop (RCL) of these proteinaceous inhibitors is crucial for protease binding and is conserved across the Pin-II family. However, the molecular mechanism and inhibitory potential of the RCL tripeptides in isolation of the native protein has remained elusive. In this study, six peptides corresponding to the RCLs of the predominant Pin-II PIs were identified, synthesized and evaluated for in vitro and in vivo inhibitory activity against serine proteases of the polyphagous insect, Helicoverpa armigera. RCL peptides with sequences PRN, PRY and TRE were found to be potent inhibitors that adversely affected the growth and development of H. armigera. The binding mechanism and differential affinity of the RCL peptides with serine proteases was delineated by crystal structures of complexes of the RCL peptides with trypsin. Residues P1 and P2 of the inhibitors play a crucial role in the interaction and specificity of these inhibitors. Important features of RCL peptides like higher inhibition of insect proteases, enhanced efficacy at alkaline gut pH, longer retention and high stability in insect gut make them suitable molecules for the development of sustainable pest management strategies for crop protection.