LONDRAVILLE, RL; KINTER, M; JURY, DR; University of Akron; Lerner Research Institute, Cleveland Clinic Foundation; University of Akron: Proteomic Analyses of Zebrafish Liver- Response to High Calorie Diet

We investigated the effects of a high-calorie diet on protein expression in zebrafish (Danio rerio). Fish were fed either 3x/day (high calorie) or 1x/3 days (low calorie) for 4 weeks. Total protein was then extracted from liver and separated by high-resolution 2-dimensional electrophoresis. With sufficient care, commercially prepared gels, and analysis software, electrophoresis results are highly repeatable and the same individual bands can be identified among gels. Several of these individual bands were characterized by in-gel trypsin digestion followed by LC-tandem MS and Maldi-TOF mass spectrometry analyses. Proteins were identified by positive match between sample and database spectra and/or primary sequence generated by interpretation of spectra. HSP 60, apolipoprotein, translationally controlled tumor protein and eukaryotic translation initiation factor 5A all increase in relative expression in liver of fish on a high-calorie diet. In addition, we identified several other proteins as landmarks in the zebrafish liver proteome, and determined partial sequence for two proteins that do not align with sequences currently in GenBank. This experiment is part of a larger project with the goal of identifying/characterizing all proteins in the zebrafish proteome.