PUBLICATION

Detection of the p53 response in zebrafish embryos using new monoclonal antibodies

Authors
Lee, K.C., Goh, W.L., Xu, M., Kua, N., Lunny, D., Wong, J.S., Coomber, D., Vojtesek, B., Lane, E.B., and Lane, D.P.
ID
ZDB-PUB-070813-16
Date
2008
Source
Oncogene   27(5): 629-640 (Journal)
Registered Authors
Keywords
zebrafish, p53, monoclonal antibody, immunohistochemistry, p53-activating agents
MeSH Terms
  • Animals
  • Antibodies, Monoclonal
  • Apoptosis
  • Embryo, Nonmammalian/metabolism
  • Gene Expression Profiling
  • Gene Expression Regulation, Developmental*
  • Immunohistochemistry/methods
  • Tumor Suppressor Protein p53/analysis
  • Tumor Suppressor Protein p53/genetics
  • Tumor Suppressor Protein p53/metabolism*
  • Zebrafish
  • Zebrafish Proteins/analysis
  • Zebrafish Proteins/metabolism*
PubMed
17684488 Full text @ Oncogene
Abstract
The zebrafish has many advantages as a vertebrate model organism and has been extensively used in the studies of development. Its potential as a model in which to study tumour suppressor and oncogene function is now being realized. Whilst in situ hybridization of mRNA has been well developed in this species to study gene expression, antibody probes are in short supply. We have, therefore, generated a panel of anti-zebrafish p53 monoclonal antibodies and used these to study the p53 response in zebrafish embryos. By immunohistochemistry, we show that the exposure of zebrafish embryos to p53-activating agents such as R-roscovitine and gamma-irradiation results in the accumulation of p53 protein in the gut epithelium, liver and pancreas. A combination of R-roscovitine and gamma-irradiation results in massive p53 induction, not only in the pharyngeal arches, gut region and liver but also in brain tissues. Induction of apoptosis and expression of p53 response genes are seen in regions that correspond to sites of p53 protein accumulation. In contrast, although zebrafish tp53(M214K) mutant embryos showed a similar accumulation of p53 protein, a complete lack of a downstream p53-dependent response was observed. In this system the p53 gene is identified as a p53-responsive gene itself. Our results demonstrate that zebrafish p53 protein can readily be induced in embryos and detected using these new antibody tools, which will increase the usefulness of zebrafish as a model in compound-based screening for novel drugs in cancer research.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping