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Originally published In Press as doi:10.1074/mcp.M700382-MCP200 on January 22, 2008.
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Molecular & Cellular Proteomics 7:981-994, 2008.
© 2008 by The American Society for Biochemistry and Molecular Biology, Inc.


Research

Analysis of the Zebrafish Proteome during Embryonic Development*,S

Margaret B. Lucitt{ddagger},§, Thomas S. Price{ddagger},§, Angel Pizarro{ddagger},§, Weichen Wu{ddagger},§, Anastasia K. Yocum§, Christoph Seiler||, Michael A. Pack||, Ian A. Blair§,**, Garret A. FitzGerald{ddagger},§,{ddagger}{ddagger} and Tilo Grosser{ddagger},§,§§

From the {ddagger} Institute for Translational Medicine and Therapeutics, Centers for Cancer Pharmacology and Excellence in Environmental Toxicology, and Departments of § Pharmacology and || Medicine, School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104

The model organism zebrafish (Danio rerio) is particularly amenable to studies deciphering regulatory genetic networks in vertebrate development, biology, and pharmacology. Unraveling the functional dynamics of such networks requires precise quantitation of protein expression during organismal growth, which is incrementally challenging with progressive complexity of the systems. In an approach toward such quantitative studies of dynamic network behavior, we applied mass spectrometric methodology and rigorous statistical analysis to create comprehensive, high quality profiles of proteins expressed at two stages of zebrafish development. Proteins of embryos 72 and 120 h postfertilization (hpf) were isolated and analyzed both by two-dimensional (2D) LC followed by ESI-MS/MS and by 2D PAGE followed by MALDI-TOF/TOF protein identification. We detected 1384 proteins from 327,906 peptide sequence identifications at 72 and 120 hpf with false identification rates of less than 1% using 2D LC-ESI-MS/MS. These included only ~30% of proteins that were identified by 2D PAGE-MALDI-TOF/TOF. Roughly 10% of all detected proteins were derived from hypothetical or predicted gene models or were entirely unannotated. Comparison of proteins expression by 2D DIGE revealed that proteins involved in energy production and transcription/translation were relatively more abundant at 72 hpf consistent with faster synthesis of cellular proteins during organismal growth at this time compared with 120 hpf. The data are accessible in a database that links protein identifications to existing resources including the Zebrafish Information Network database. This new resource should facilitate the selection of candidate proteins for targeted quantitation and refine systematic genetic network analysis in vertebrate development and biology.


{ddagger}{ddagger} The McNeil Professor in Translational Medicine and Therapeutics. To whom correspondence may be addressed: Inst. for Translational Medicine and Therapeutics, University of Pennsylvania, 153 Johnson Pavilion, 3620 Hamilton Walk, Philadelphia, PA 19104-6084. Tel.: 215-898-1184; E-mail: garret{at}spirit.gcrc.upenn.edu

§§ To whom correspondence may be addressed: Inst. for Translational Medicine and Therapeutics, University of Pennsylvania, 153 Johnson Pavilion, 3620 Hamilton Walk, Philadelphia, PA 19104-6084. Tel.: 215-898-1184; E-mail: tilo{at}itmat.upenn.edu







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