Identifying candidate proteins that undergo activity-dependent synapse to nucleus translocation in hippocampal neurons
Tay, Eden Xin Jie
Date of Issue2017-08-31
Lee Kong Chian School of Medicine (LKCMedicine)
Previous studies have explored the mechanisms that proteins from the post-synaptic terminal utilise during retrograde transport to the nucleus (1,2) and their role on learning and memory formation in neurons (3–5), but not much is known about proteins isolated from the pre-synaptic terminal(6). Here, we investigate the functional capabilities of a purified list of isolated proteins from the presynaptic terminal, especially looking out for transcriptional regulation and modulation. We also look for possible nuclear translocation in the form of nuclear localising protein motifs in their sequences. At the same time, we compare the efficacy of two different antibodies (Rabbit Polyclonal Anti-GFP antibody vs Mouse Monoclonal Anti-GFP antibody) in our proteomic quantification protocol, INTACT, for use in quantitative analysis of possible activity-dependent neuronal nuclear translocation of proteins from the pre-synaptic terminal. We identified 5 candidate proteins with high likelihood of nuclear translocation and transcriptional activity for further testing. We also managed to show that the Rabbit Polycloncal Anti-GFP antibody is more efficient at immunoprecipitation of Sun1GFP expressing activated nuclei.
Final Year Project (FYP)