In Vivo Imaging of Embryonic Vascular Development Using Transgenic Zebrafish

Dev Biol. 2002 Aug 15;248(2):307-18. doi: 10.1006/dbio.2002.0711.

Abstract

In this study we describe a model system that allows continuous in vivo observation of the vertebrate embryonic vasculature. We find that the zebrafish fli1 promoter is able to drive expression of enhanced green fluorescent protein (EGFP) in all blood vessels throughout embryogenesis. We demonstrate the utility of vascular-specific transgenic zebrafish in conjunction with time-lapse multiphoton laser scanning microscopy by directly observing angiogenesis within the brain of developing embryos. Our images reveal that blood vessels undergoing active angiogenic growth display extensive filopodial activity and pathfinding behavior similar to that of neuronal growth cones. We further show, using the zebrafish mindbomb mutant as an example, that the expression of EGFP within developing blood vessels permits detailed analysis of vascular defects associated with genetic mutations. Thus, these transgenic lines allow detailed analysis of both wild type and mutant embryonic vasculature and, together with the ability to perform large scale forward-genetic screens in zebrafish, will facilitate identification of new mutants affecting vascular development.

Publication types

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

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Blood Vessels / embryology*
  • Brain / blood supply*
  • Brain / embryology*
  • Genes, Reporter / genetics
  • Germ-Line Mutation / genetics
  • Green Fluorescent Proteins
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Microscopy, Confocal / methods*
  • Promoter Regions, Genetic / genetics
  • Time Factors
  • Transgenes / genetics*
  • Zebrafish / embryology*
  • Zebrafish / genetics*
  • Zebrafish Proteins / genetics
  • Zebrafish Proteins / metabolism

Substances

  • Luminescent Proteins
  • Zebrafish Proteins
  • Green Fluorescent Proteins