PUBLICATION

SCL-GFP transgenic zebrafish: In vivo imaging of blood and endothelial development and identification of the initial site of definitive hematopoiesis

Authors
Zhang, X.Y., and Rodaway, A.R.
ID
ZDB-PUB-070625-20
Date
2007
Source
Developmental Biology   307(2): 179-194 (Journal)
Registered Authors
Rodaway, Adam, Zhang, Xiang Yi
Keywords
TAL1, AML1, Globin, Fugu, Pufferfish, Recombinase, Recombineering, Transgenesis, mRFP, Vasculature, Haematopoiesis
MeSH Terms
  • Zebrafish/blood
  • Zebrafish/genetics
  • Zebrafish/growth & development*
  • Animals
  • Proto-Oncogene Proteins/genetics
  • Proto-Oncogene Proteins/physiology*
  • Base Sequence
  • Hematopoiesis/genetics
  • Hematopoiesis/physiology*
  • Gene Expression Regulation, Developmental
  • Basic Helix-Loop-Helix Transcription Factors/genetics
  • Basic Helix-Loop-Helix Transcription Factors/physiology*
  • Cell Movement
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/physiology*
  • Recombinant Proteins/genetics
  • Green Fluorescent Proteins/genetics
  • Animals, Genetically Modified
  • DNA Primers/genetics
  • Hematopoietic Stem Cells/cytology
(all 20)
PubMed
17559829 Full text @ Dev. Biol.
Abstract
The bHLH transcription factor SCL plays a central role in the generation of hematopoietic cells in vertebrates. We modified a PAC containing the whole zebrafish scl locus, inserting GFP into the first coding exon of scl. In germline-transgenic zebrafish generated using this construct, GFP expression completely recapitulates the endogenous expression of scl in blood, endothelium and CNS. We performed in vivo timelapse imaging of blood and endothelial precursor migration at the single-cell level and show that these cells migrate from the posterior lateral plate mesoderm to their site of differentiation in the intermediate cell mass. The anterior lateral plate domain of GFP expression gives rise to primitive macrophages and the blood vessels of the head. In later embryos, GFP expression identifies clusters of hematopoietic cells that develop between the dorsal aorta and posterior cardinal veins after primitive erythrocytes have entered circulation. Two treatments that block definitive hematopoiesis (treatment with dioxin (TCDD), and injection of an antisense morpholino oligonucleotide targeted to runx1) ablate these hematopoietic clusters. This indicates that these clusters represent the first site of definitive hematopoiesis in zebrafish. This site is anatomically homologous to the proposed source of hematopoietic stem cells in amniotes, the aorta-gonad-mesonephros (AGM) region. A second transgenic line, containing the promoter of scl driving GFP, lacks expression in the definitive clusters.
Genes / Markers
Figures
Figure Gallery (7 images)
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Expression
Phenotype
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
zf254TgTransgenic Insertion
    zf255TgTransgenic Insertion
      1 - 2 of 2
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      Human Disease / Model
      No data available
      Sequence Targeting Reagents
      Target Reagent Reagent Type
      runx1MO1-runx1MRPHLNO
      1 - 1 of 1
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      Fish
      Fish
      WT
      WT + MO1-runx1
      zf254Tg
      zf255Tg
      1 - 4 of 4
      Show
      Antibodies
      No data available
      Orthology
      No data available
      Engineered Foreign Genes
      Marker Marker Type Name
      EGFPEFGEGFP
      1 - 1 of 1
      Show
      Mapping
      No data available
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      Citation
      Zhang, X.Y., and Rodaway, A.R. (2007) SCL-GFP transgenic zebrafish: In vivo imaging of blood and endothelial development and identification of the initial site of definitive hematopoiesis. Developmental Biology. 307(2):179-194.