Identifying candidate proteins that undergo activity-dependent synapse to nucleus translocation in hippocampal neurons
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). H...
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| Format: | Final Year Project |
| Language: | English |
| Published: |
2017
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| Subjects: | |
| Online Access: | http://hdl.handle.net/10356/72642 |
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| Institution: | Nanyang Technological University |
| Language: | English |
| Summary: | 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. |
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