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P1‐016: A late expression of brain‐derived neurotrophic factor is required for the synaptic remodeling in the persistence of fear memory
Author(s) -
Ou LiChin,
Yeh ShiuHwa,
Gean PoWu
Publication year - 2009
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1016/j.jalz.2009.04.019
Subject(s) - tropomyosin receptor kinase b , fear conditioning , amygdala , neurotrophic factors , brain derived neurotrophic factor , conditioning , synaptic plasticity , psychology , fear processing in the brain , neuroscience , neuroplasticity , endocrinology , medicine , receptor , statistics , mathematics
Background: Genetic analysis has been a potent tool in understanding processes involved in development and disease for more than a decade. However, differences in mRNA expression are not a reliable predictor of changes in protein expression (Chen et al., 2002; Greenbaum et al., 2003; Gygi et al., 1999). As the methods of mass spectrometry and the possibilities to analyze data advance the alternative of investigating global proteoms becomes achievable. Our aim is to study changes in protein expression during Alzheimer’s disease (AD) using zebrafish as a model system. Zebrafish has several advantages over e.g., mice in that they are cheap, easily manipulated both genetically and with chemical compounds and can also be monitored during development. Methods: We are currently developing both methods to get a fast and crude proteome scan of the zebrafish brain (the shotgun approach), and also more detailed and time-consuming methods involving prefractionation of proteins with SDS-PAGE gels followed by enzymatic digestion and LC-MS/MS identification. Results: Currently we have been able to identify over 200 brain-associated proteins from a brain extract, several of which have not been reported earlier in the literature. Proteomics is often very time consuming and as a result few replicates are made. The widely used method to detect differences in protein level using SILAC (stable isotope labeling with amino acids in cell culture) was recently applied on mouse to compare protein levels of knock out mice in vivo (Kruger et al., 2008). We use this method on zebrafish and preliminary data show a 15% labeling of brain proteins after feeding adult fish heavy 13C6-Lysine for 4 months. Brain has a low turn over and the incorporation is therefore slower than in other tissues, like gut and blood, which are expected to have a higher incorporation level. We are currently extending the labeling over generations and expect to get a near 100% labeled fish. Conclusions: We hope to measure the brain proteome and the change in specific proteins in the zebrafish brain after silencing genes implicated in AD. The relatively fast procedure will facilitate analyses of multiple samples and hence increase the reliability of the results.