Identification of regulators of germ layer morphogenesis using proteomics in zebrafish Journal Article


Author(s): Link, Vinzenz; Carvalho, Lara; Castanon, Irinka; Stockinger, Petra; Shevchenko, Andrej; Heisenberg, Carl-Philipp
Article Title: Identification of regulators of germ layer morphogenesis using proteomics in zebrafish
Affiliation
Abstract: During vertebrate gastrulation, a well-orchestrated series of morphogenetic changes leads to the formation of the three germ layers: the ectoderm, mesoderm and endoderm. The analysis of gene expression patterns during gastrulation has been central to the identification of genes involved in germ layer formation. However, many proteins are regulated on a translational or post-translational level and are thus undetectable by gene expression analysis. Therefore, we developed a 2D-gel-based comparative proteomic approach to target proteins involved in germ layer morphogenesis during zebrafish gastrulation. Proteomes of ectodermal and mesendodermal progenitor cells were compared and 35 significantly regulated proteins were identified by mass spectrometry, including several proteins with predicted functions in cytoskeletal organization. A comparison of our proteomic results with data obtained in an accompanying microarray-based gene expression analysis revealed no significant overlap, confirming the complementary nature of proteomics and transcriptomics. The regulation of ezrin2, which was identified based on a reduction in spot intensity in mesendodermal cells, was independently validated. Furthermore, we show that ezrin2 is activated by phosphorylation in mesendodermal cells and is required for proper germ layer morphogenesis. We demonstrate the feasibility of proteomics in zebrafish, concluding that proteomics is a valuable tool for analysis of early development.
Keywords: Proteomics; Gastrulation; Zebrafish; Ezrin
Journal Title: Journal of Cell Science
Volume: 119
Issue 10
ISSN: 1477-9137
Publisher: Company of Biologists  
Date Published: 2006-05-15
Start Page: 2073
End Page: 2083
DOI: 10.1242/jcs.02928
Open access: no