PUBLICATION
Identification of regulators of germ layer morphogenesis using proteomics in zebrafish
- Authors
- Link, V., Carvalho, L., Castanon, I., Stockinger, P., Shevchenko, A., and Heisenberg, C.P.
- ID
- ZDB-PUB-060501-10
- Date
- 2006
- Source
- Journal of Cell Science 119(10): 2073-2083 (Journal)
- Registered Authors
- Castanon-Ortega, Mirna-Irinka, Heisenberg, Carl-Philipp, Link, Vinzenz
- Keywords
- Proteomics, Ezrin, Gastrulation, Zebrafish
- MeSH Terms
-
- Animals
- Cytoskeletal Proteins/biosynthesis
- Cytoskeletal Proteins/genetics
- Cytoskeletal Proteins/physiology
- Gene Expression Profiling
- Gene Expression Regulation, Developmental
- Genomics/methods
- Germ Layers/cytology
- Germ Layers/metabolism
- Germ Layers/physiology*
- Mass Spectrometry/methods
- Microarray Analysis
- Morphogenesis
- Oligonucleotides, Antisense/genetics
- Proteomics/methods*
- Zebrafish/embryology*
- Zebrafish/genetics
- Zebrafish/metabolism
- Zebrafish Proteins/biosynthesis
- Zebrafish Proteins/genetics
- Zebrafish Proteins/physiology*
- PubMed
- 16638810 Full text @ J. Cell Sci.
Citation
Link, V., Carvalho, L., Castanon, I., Stockinger, P., Shevchenko, A., and Heisenberg, C.P. (2006) Identification of regulators of germ layer morphogenesis using proteomics in zebrafish. Journal of Cell Science. 119(10):2073-2083.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping