PUBLICATION

A genetic screen for vascular mutants in zebrafish reveals dynamic roles for Vegf/Plcg1 signaling during artery development

Authors

Covassin, L.D., Siekmann, A.F., Kacergis, M.C., Laver, E., Moore, J.C., Villefranc, J.A., Weinstein, B.M., and Lawson, N.D.

ID

ZDB-PUB-090310-21

Date

2009

Source

Developmental Biology 329(2): 212-226 (Journal)

Registered Authors

Covassin, Laurence, Kacergis, Michael C., Laver, Elizabeth, Lawson, Nathan, Villefranc, Jacque, Weinstein, Brant M.

Keywords

Zebrafish, Vascular, Genetic screen, Vegf

MeSH Terms

Signal Transduction*
Zebrafish/embryology*
Zebrafish/genetics
Base Sequence
Mutation*
Amino Acid Sequence
Mosaicism
Animals, Genetically Modified
Phospholipase C gamma/chemistry
Phospholipase C gamma/physiology*
Arteries/embryology*
Female
Male
Vascular Endothelial Growth Factor A/chemistry
Vascular Endothelial Growth Factor A/physiology*
Sequence Homology, Amino Acid
Molecular Sequence Data
DNA Primers
Animals

(all 19)

PubMed

19269286 Full text @ Dev. Biol.

Abstract

In this work we describe a forward genetic approach to identify mutations that affect blood vessel development in the zebrafish. By applying a haploid screening strategy in a transgenic background that allows direct visualization of blood vessels, it was possible to identify several classes of mutant vascular phenotypes. Subsequent characterization of mutant lines revealed that defects in Vascular endothelial growth factor (Vegf) signaling specifically affected artery development. Comparison of phenotypes associated with different mutations within a functional zebrafish Vegf receptor-2 ortholog (referred to as kdr-like, kdrl) revealed surprisingly varied effects on vascular development. In parallel, we identified an allelic series of mutations in phospholipase c gamma 1 (plcg1). Together with in vivo structure-function analysis, our results suggest a requirement for Plcg1 catalytic activity downstream of receptor tyrosine kinases. We further find that embryos lacking both maternal and zygotic plcg1 display more severe defects in artery differentiation but are otherwise similar to zygotic mutants. Finally, we demonstrate through mosaic analysis that plcg1 functions autonomously in endothelial cells. Together our genetic analyses suggest that Vegf/Plcg1 signaling acts at multiple time points and in different signaling contexts to mediate distinct aspects of artery development.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Qualifier	Anatomy	Assay	Figure
dll4	plcg1^y18/+; y1Tg/+	standard conditions	Prim-15	Not Detected	dorsal aorta	ISH	Fig. 6
dll4	plcg1^y18/y18	standard conditions	Prim-15	Not Detected	dorsal aorta	ISH	Fig. 6
efnb2a	plcg1^y13/y13; y1Tg	standard conditions	Prim-15	Not Detected	dorsal aorta	ISH	Fig. 6
efnb2a	plcg1^y16/y16; y1Tg	standard conditions	Prim-15		dorsal aorta	ISH	Fig. 6
efnb2a	plcg1^y18/y18	standard conditions	Prim-15	Not Detected	dorsal aorta	ISH	Fig. 6
efnb2a	plcg1^y18/y18; y1Tg	standard conditions	Prim-15		dorsal aorta	ISH	Fig. 6
efnb2a	WT	standard conditions	Prim-15		dorsal aorta	ISH	Fig. 6
EGFP	kdrl^um19/um19; y1Tg	standard conditions	Prim-15		dorsal aorta	IFL	Fig. 2
EGFP	kdrl^um19/um19; y1Tg	standard conditions	Prim-15		intersegmental vessel	IFL	Fig. 2
EGFP	kdrl^um6/um6; y1Tg	standard conditions	Prim-15		dorsal aorta	IFL	Fig. 2

1 - 10 of 25

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
kdrl^um19/um19; y1Tg	standard conditions	Prim-15	angiogenesis disrupted, abnormal	Fig. 2
kdrl^um19/um19; y1Tg	standard conditions	Prim-15	dorsal longitudinal anastomotic vessel aplastic, abnormal	Fig. 2
kdrl^um19/um19; y1Tg	standard conditions	Prim-15	intersegmental artery decreased length, abnormal	Fig. 2
kdrl^um19/um19; y1Tg	standard conditions	Long-pec	blood circulation process quality, normal	Fig. 3
kdrl^um19/um19; y1Tg	standard conditions	Long-pec	cranial vasculature broken, abnormal	Fig. 3
kdrl^um19/um19; y1Tg	standard conditions	Long-pec	dorsal aorta morphology, abnormal	Fig. 3
kdrl^um19/um19; y1Tg	standard conditions	Long-pec	intersegmental artery decreased length, abnormal	Fig. 3
kdrl^um19/um19; y1Tg	standard conditions	Long-pec	pericardium morphology, normal	Fig. 3
kdrl^um19/um19; y1Tg	standard conditions	Long-pec	posterior cardinal vein morphology, abnormal	Fig. 3
kdrl^um6/+; y1Tg	standard conditions	Long-pec	cranial vasculature broken, abnormal	Fig. 3

1 - 10 of 40

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
um6		Point Mutation	kdrl
um7		Unknown	plxnd1
um13Tg	Tg(fli1.ep:DsRedEx)	Transgenic Insertion
um19		Small Deletion	kdrl
y1Tg	Tg(fli1:EGFP)	Transgenic Insertion
y10		Point Mutation	plcg1
y11		Insertion	etsrp
y13		Point Mutation	plcg1
y15		Point Mutation	plcg1
y16		Point Mutation	plcg1

1 - 10 of 13

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Human Disease / Model

Sequence Targeting Reagents

Fish

Fish
etsrp^y11/+; y1Tg/y1Tg
etsrp^y11/y11; y1Tg
kdrl^um6/+
kdrl^um6/+; y1Tg
kdrl^um6/+; y1Tg/y1Tg
kdrl^um6/um6; y1Tg
kdrl^um19/um19; y1Tg
kdrl^y17/+; y1Tg/y1Tg
kdrl^y17/y17; y1Tg
plcg1^y10/+; y1Tg/y1Tg

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Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP

Mapping

Covassin et al., 2009 ZDB-PUB-090310-21 PMID:19269286

A genetic screen for vascular mutants in zebrafish reveals dynamic roles for Vegf/Plcg1 signaling during artery development

Covassin et al., 2009

ZDB-PUB-090310-21
PMID:19269286