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Proteomics of early zebrafish embryos

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BioMed Central
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PMC
Keywords
  • Methodology Article
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  • Biology

Abstract

1471-213X-6-1.fm ral ss BioMed CentBMC Developmental Biology Open AcceMethodology article Proteomics of early zebrafish embryos Vinzenz Link, Andrej Shevchenko and Carl-Philipp Heisenberg* Address: Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauer Str. 108, 01307 Dresden, Germany Email: Vinzenz Link - [email protected]; Andrej Shevchenko - [email protected]; Carl-Philipp Heisenberg* - [email protected] * Corresponding author Abstract Background: Zebrafish (D. rerio) has become a powerful and widely used model system for the analysis of vertebrate embryogenesis and organ development. While genetic methods are readily available in zebrafish, protocols for two dimensional (2D) gel electrophoresis and proteomics have yet to be developed. Results: As a prerequisite to carry out proteomic experiments with early zebrafish embryos, we developed a method to efficiently remove the yolk from large batches of embryos. This method enabled high resolution 2D gel electrophoresis and improved Western blotting considerably. Here, we provide detailed protocols for proteomics in zebrafish from sample preparation to mass spectrometry (MS), including a comparison of databases for MS identification of zebrafish proteins. Conclusion: The provided protocols for proteomic analysis of early embryos enable research to be taken in novel directions in embryogenesis. Background The zebrafish has become a widely used vertebrate model system for which a large tool-box of genetic and cell bio- logical methods has been established [1,2]. Research using zebrafish is further supported by the zebrafish sequencing project, which has facilitated the generation of microarrays for large scale expression profiling. It has been proposed that proteomics should complement the genome-wide expression profiling [3]. However, a major obstacle in the application of proteomics has been the high proportion of yolk proteins in early embryos. Pro- teomic studies in zebrafish have therefore be

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