Distinct Requirements for Zebrafish Angiogenesis Revealed by a VEGF-A Morphant

Yeast. 2000 Dec;17(4):294-301. doi: 10.1002/1097-0061(200012)17:4<294::AID-YEA54>3.0.CO;2-5.

Abstract

Angiogenesis is a fundamental vertebrate developmental process that requires signalling by the secreted protein vascular endothelial growth factor-A (VEGF-A). VEGF-A functions in the development of embryonic structures, during tissue remodelling and for the growth of tumour-induced vasculature. The study of the role of VEGF-A during normal development has been significantly complicated by the dominant, haplo-insufficient nature of VEGF-A-targeted mutations in mice. We have used morpholino-based targeted gene knock-down technology to generate a zebrafish VEGF-A morphant loss of function model. Zebrafish VEGF-A morphant embryos develop with an enlarged pericardium and with major blood vessel deficiencies. Morphological assessment at 2 days of development indicates a nearly complete absence of both axial and intersegmental vasculature, with no or reduced numbers of circulating red blood cells. Molecular analysis using the endothelial markers fli-1 and flk-1 at 1 day of development demonstrates a fundamental distinction between VEGF-A requirements for axial and intersegmental vascular structure specification. VEGF-A is not required for the initial establishment of axial vasculature patterning, whereas all development of intersegmental vasculature is dependent on VEGF-A signalling. The zebrafish thus serves as a quality model for the study of conserved vertebrate angiogenesis processes during embryonic development.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Blood Vessels / embryology*
  • Body Patterning
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Embryonic Development
  • Endothelial Growth Factors / genetics*
  • Endothelial Growth Factors / physiology*
  • Gene Expression
  • Gene Targeting*
  • Morpholines
  • Mutation
  • Neovascularization, Physiologic*
  • Oligonucleotides, Antisense
  • Phenotype
  • Proto-Oncogene Protein c-fli-1
  • Proto-Oncogene Proteins*
  • Receptor Protein-Tyrosine Kinases / genetics
  • Receptor Protein-Tyrosine Kinases / metabolism
  • Receptors, Growth Factor / genetics
  • Receptors, Growth Factor / metabolism
  • Receptors, Vascular Endothelial Growth Factor
  • Signal Transduction
  • Trans-Activators / genetics
  • Trans-Activators / metabolism
  • Vascular Endothelial Growth Factor A
  • Zebrafish / embryology*
  • Zebrafish / genetics*

Substances

  • DNA-Binding Proteins
  • Endothelial Growth Factors
  • Morpholines
  • Oligonucleotides, Antisense
  • Proto-Oncogene Protein c-fli-1
  • Proto-Oncogene Proteins
  • Receptors, Growth Factor
  • Trans-Activators
  • Vascular Endothelial Growth Factor A
  • Receptor Protein-Tyrosine Kinases
  • Receptors, Vascular Endothelial Growth Factor