In vivo imaging of embryonic vascular development using transgenic zebrafish.

By Nathan D. Lawson, B. Weinstein. Developmental Biology, 2002

2,179 citations recorded by the source corpus.

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.

Subfields: Cell Biology, Cellular and Molecular Neuroscience, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/57dba18ba3fa3146c0b213a02bdb8efff49da5ce

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Citations in the corpus

SIRT1 controls endothelial angiogenic functions during vascular growth. (2007, methodology): “Embryos of the tg(fli1:eGFP) line (Lawson and Weinstein 2002) characterized by fluorescent endothelial cells were used to ease the analysis of the phenotype.”
A dynamic epicardial injury response supports progenitor cell activity during zebrafish heart regeneration. (2006, positive): “Over the past several years, large-scale genetic screens (Chen et al., 1996; Stainier et al., 1996), transgenic reporter strains ( Lawson and Weinstein, 2002 ), and lineage studies (Keegan et al., 2005) have revealed critical aspects of cardiovascular development in zebrafish embryos.”
Targeted gene inactivation in zebrafish using engineered zinc-finger nucleases (2008, background): “These modified Fok I nuclease domains contain previously described mutations at the dimerization interface that reduce their propensity to homodimeriz”
Zebrafish as a model vertebrate for investigating chemical toxicity. (2005, background): “1), which permit visualization of endothelial cells of different vascular beds for the study of toxicants that disrupt cardiovascular development (Bello et al., 2004; Lawson and Weinstein, 2002).”
Cloning of hif-1α and hif-2α and mRNA expression pattern during development in zebrafish (2007, background): “cells under control of the Xi1 promoter ( Lawson and Weinstein, 2002 ).”
Cellular and molecular analyses of vascular tube and lumen formation in zebrafish (2005, methodology): “5199”
Identification of RNF213 as a Susceptibility Gene for Moyamoya Disease and Its Possible Role in Vascular Development (2011, methodology): “Knockdown of RNF213-a and RNF213-b expression was achieved by injection of a specific morpholino (MO; Gene Tools) into 1- to 8-cell stage embryos, as described previously [28].”
Role of PFKFB3-driven glycolysis in vessel sprouting. (2013, positive): “Animal Models of Angiogenesis Tg(fli1:EGFP) zebrafish (Lawson and Weinstein, 2002) were subjected to Tol2-based transgenic overexpression.”
Reverse genetic screening reveals poor correlation between Morpholino-induced and mutant phenotypes in zebrafish (2014, methodology): “The formation of these vessels has been used to screen for genes required for angiogenesis (for examples, see Covassin et al., 2009; Isogai et al., 2003; Lawson and Weinstein, 2002).”
Endothelial cells dynamically compete for the tip cell position during angiogenic sprouting (2010, neutral): “, denoted as kdrl:mCherry; Tg ( fli1a : eGFP) , denoted as flia:eGFP ; Tg ( fli1a:NLS–GFP)”