School of Science Division of Life Science 20 Cell Surface Receptor Interactome Related to Forgetting Supervisor: HIRANO Yukinori / LIFS Student: LANG Xuan / BIOT-IRE ZHANG Shunxi / BIOT Course: UROP 1000, Summer UROP 1100, Summer Active forgetting, referring to the brain’s active mechanisms for suppressing specific memories, allows flexibility and efficiency in memory regulation. Building on Professor Yukinori Hirano’s discovery of a particular gene-expression-based pathway of forgetting, this project investigates the role of Kek2, a synaptic plasticity regulator in dopamine-dependent memory modulation. Previous UROP students have planned to construct 18 expression plasmids which encode candidate proteins from proteomic data. Based on their direction, we continued amplifying remaining target genes, especially NT1, the Drosophila Neurotrophin. However, challenges like faint band and smeared bands exist during the amplification of NT1, due to its low expression and the imperfect quality of cDNA. Meanwhile, we successfully amplified and construct the plasmid of PdhRA, bw, and CG17600. We further co-transfected S2 cells with HA-tagged candidate proteins and 6×FLAG-tagged kek2 plasmids for immunoprecipitation (IP) and western blot (WB) analysis. Although the transfection turns out to be efficacious, the results of immunoprecipitation remain unreliable, only indicating the possibility of CG17600 interacting with kek2. Our future work will focus on the nested PCR for unamplified genes, and Oligo-dT primers purification of pupae mRNA of drosophila, followed by reverse transcription to get higher quality of cDNA. Also, more solid IP and WB analysis will be conveyed to complete the investigation on PdhRA, bw, and CG17600. Cell Surface Receptor Interactome Related to Forgetting Supervisor: HIRANO Yukinori / LIFS Student: SAHA Anya / BIOT Course: UROP 1000, Summer The mechanism through which anxiety affects internal state and behavior is an area still not completely understood. Our groundwork was set by Professor Hirano’s discovery of how Allostatin-A receptors perform neuropeptide signaling to induce claustrophobic behavior as a response to stress, unlike normal fear conditioning which uses dopamine receptors. Neuropeptide signaling is more prolonged, which explains the persistence of behavioral change. While we began by investigating the cell surface receptor interactome related to forgetting, upon learning of this discovery, we started investigating the interactome of AllostatinA neuropeptide signaling receptors instead. In this project, we look to investigate if the proteins interacting with Allostatin-A which the lab team identified also interact similarly with other Allostatin signaling receptors.
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