PUBLICATION

Dominant-interfering C/EBPalpha stimulates primitive erythropoiesis in zebrafish

Authors
Liu, T.X., Rhodes, J., Deng, M., Hsu, K., Radomska, H.S., Kanki, J.P., Tenen, D.G., and Look, A.T.
ID
ZDB-PUB-070210-43
Date
2007
Source
Experimental hematology   35(2): 230-239 (Journal)
Registered Authors
Deng, Min, Hsu, Karl, Kanki, John, Look, A. Thomas, Rhodes, Jennifer
Keywords
none
MeSH Terms
  • Animals
  • Base Sequence
  • Blood Vessels/embryology
  • Blood Vessels/metabolism
  • CCAAT-Enhancer-Binding Protein-alpha/genetics*
  • CCAAT-Enhancer-Binding Protein-alpha/metabolism
  • DNA, Complementary/genetics
  • Embryonic Development/genetics
  • Embryonic Development/physiology
  • Erythropoiesis/genetics*
  • Gene Deletion
  • Gene Expression Regulation, Developmental*
  • Genes, Dominant*
  • Hematopoietic Stem Cells/cytology
  • Hematopoietic Stem Cells/metabolism
  • Humans
  • Molecular Sequence Data
  • Mutation
  • Promoter Regions, Genetic/genetics
  • Promoter Regions, Genetic/physiology
  • RNA, Messenger
  • Reverse Transcriptase Polymerase Chain Reaction/methods
  • Sequence Analysis, DNA
  • Translocation, Genetic/genetics
  • Translocation, Genetic/physiology
  • Transplantation, Heterologous
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/metabolism
PubMed
17258072 Full text @ Exp. Hematol.
Abstract
OBJECTIVE: We investigated the role of CCAAT enhancer-binding protein-alpha (C/EBPalpha) during zebrafish embryonic blood development. METHODS: Whole-mount mRNA in situ hybridization was performed to determine the spatio-temporal expression pattern of zebrafish cebpa in developing hematopoietic progenitors. A deletion mutation of cebpa (zD420), which mimics the human dominant-negative mutations of C/EBPalpha, was transfected into CV1 cell line to evaluate its transcriptional activity in vitro and injected into zebrafish embryos at the one- to two-cell stage to examine its effects on primitive hematopoiesis during early zebrafish development. RESULTS: Zebrafish cebpa is expressed in the anterior and posterior lateral plate mesoderm at 12 hours postfertilization, along with scl, pu.1, and gata1 in developing hematopoietic progenitors. In vitro, the deletion mutation of cebpa (zD420) prevents expression of the full-length protein, allowing the expression of truncated isoforms from internal translational initiation sites. As in the human, the truncated zebrafish C/EBPalpha proteins did not activate the expression of known target granulocytic genes, and in fact suppressed transactivation that was induced in vitro by the full-length protein. Forced expression of the zD420 mRNA in zebrafish embryos led to an expansion of primitive erythropoiesis, without a discernible effect on granulopoiesis. CONCLUSION: Expression of the truncated isoforms of cebpa alters the developmental pattern of hematopoietic progenitor cells during embryogenesis.
Genes / Markers
Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes