PUBLICATION

Vegfa signals through ERK to promote angiogenesis, but not artery differentiation

Authors

Shin, M., Beane, T., Quillien, A., Male, I., Zhu, L.J., Lawson, N.D.

ID

ZDB-PUB-160901-3

Date

2016

Source

Development (Cambridge, England) 143(20): 3796-3805 (Journal)

Registered Authors

Lawson, Nathan, Shin, Masahiro

Keywords

ERK, Vegfa, Angiogenesis, Artery differentiation, Zebrafish

MeSH Terms

In Situ Hybridization
Cell Differentiation/genetics
Cell Differentiation/physiology
Vascular Endothelial Growth Factor A/genetics
Vascular Endothelial Growth Factor A/metabolism*
Animals
Animals, Genetically Modified
Zebrafish
Arteries/cytology*
Arteries/metabolism
Vascular Endothelial Growth Factor Receptor-3/genetics
Vascular Endothelial Growth Factor Receptor-3/metabolism
Extracellular Signal-Regulated MAP Kinases/genetics
Extracellular Signal-Regulated MAP Kinases/metabolism*
Blotting, Western
Signal Transduction/genetics
Signal Transduction/physiology
Neovascularization, Physiologic/genetics
Neovascularization, Physiologic/physiology
Receptors, Notch/genetics
Receptors, Notch/metabolism

(all 21)

PubMed

27578780 Full text @ Development

Abstract

Vascular endothelial growth factor a (Vegfa) is essential for blood vessel formation and can induce activation of numerous signaling effectors in endothelial cells. However, it is unclear how and where these function in developmental contexts during vascular morphogenesis. To address this issue, we have visualized activation of presumptive Vegfa effectors at single cell resolution in zebrafish blood vessels. From these studies, we find that phosphorylation of the serine/threonine kinase ERK (pERK) preferentially occurs in endothelial cells undergoing angiogenesis, but not in committed arterial endothelial cells. pERK in endothelial cells was ectopically induced by Vegfa and lost in Vegfa signaling mutants. Both chemical and endothelial autonomous inhibition of ERK prevented endothelial sprouting, but did not prevent initial artery differentiation. Timed chemical inhibition during angiogenesis caused a loss of genes implicated in coordinating tip/stalk cell behaviors, including flt4 and, at later stages, dll4 ERK inhibition also blocked excessive angiogenesis and ectopic flt4 expression in Notch-deficient blood vessels. Together, these studies implicate ERK as a specific effector of Vegfa signaling in the induction of angiogenic genes during sprouting.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Qualifier	Anatomy	Assay	Figure
dll4	WT	control	Prim-5		dorsal aorta	ISH	Fig. S4
dll4	WT	chemical treatment by environment: semaxanib	Prim-5	Not Detected	dorsal aorta	ISH	Fig. S4
dll4	WT	chemical treatment by environment: SL-327	Prim-5		dorsal aorta	ISH	Fig. S4
dll4	WT	control	26+ somites		endothelial cell	ISH	Fig. 5
dll4	WT	chemical treatment by environment: SL-327	26+ somites		endothelial cell	ISH	Fig. 5
dll4	WT	control	Prim-5		intersegmental vessel	ISH	Fig. 5
dll4	WT	chemical treatment by environment: SL-327	Prim-5		intersegmental vessel	ISH	Fig. 5
dll4	WT	chemical treatment by environment: SL-327	Prim-5		intersegmental vessel	ISH	Fig. S4
dll4	WT	chemical treatment by environment: semaxanib	Prim-5	Not Detected	intersegmental vessel	ISH	Fig. S4
dll4	WT	control	Prim-5		intersegmental vessel	ISH	Fig. S4

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	chemical treatment by environment: SL-327	26+ somites	endothelial cell flt4 expression decreased amount, abnormal	Fig. 5
WT	chemical treatment by environment: SL-327	Prim-5	dorsal aorta dll4 expression decreased amount, abnormal	Fig. S4
WT	chemical treatment by environment: SL-327	Prim-5	dorsal aorta efnb2a expression decreased amount, abnormal	Fig. 6
WT	chemical treatment by environment: SL-327	Prim-5	dorsal aorta notch1b expression decreased amount, abnormal	Fig. 6
WT	chemical treatment by environment: SL-327	Prim-5	intermediate cell mass of mesoderm necrotic, abnormal	Fig. 3
WT	chemical treatment by environment: SL-327	Prim-5	intersegmental vessel dll4 expression decreased amount, abnormal	Fig. S4
WT	chemical treatment by environment: SL-327	Prim-5	intersegmental vessel dll4 expression decreased amount, abnormal	Fig. 5
WT	chemical treatment by environment: SL-327	Prim-5	intersegmental vessel flt4 expression decreased amount, abnormal	Fig. S4
WT	chemical treatment by environment: SL-327	Prim-5	intersegmental vessel kdrl expression decreased amount, abnormal	Fig. S4
WT	chemical treatment by environment: SL-327	Prim-5	intersegmental vessel endothelial cell flt4 expression decreased amount, abnormal	Fig. 5

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
la116Tg	Tg(kdrl:GFP)	Transgenic Insertion
mu101Tg	TgBAC(cdh5:GAL4FF)	Transgenic Insertion
nkuasgfp1aTg	Tg(5xUAS:EGFP)	Transgenic Insertion
ta52b		Point Mutation	mib1
um19		Small Deletion	kdrl
um131		Indel	flt4
um151Tg	Tg(UAS:NLS-Tomato-2A-dusp6)	Transgenic Insertion
um152Tg	Tg(UAS:NLS-Tomato)	Transgenic Insertion
y1Tg	Tg(fli1:EGFP)	Transgenic Insertion
y13		Point Mutation	plcg1

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
dll4	MO2-dll4	MRPHLNO

Fish

Fish
kdrl^y17; flt4^um131/um131
kdrl^y17/y17; y1Tg
la116Tg
la116Tg + MO2-dll4
mib1^ta52b/ta52b
mu101Tg; nkuasgfp1aTg; um151Tg
mu101Tg; nkuasgfp1aTg; um152Tg
plcg1^y13/y13; y1Tg
WT
y1Tg

Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab1-efnb2	polyclonal	efnb2a	IgG	Goat
Ab1-myc	monoclonal		IgG1	Mouse
Ab1-phospho-(ser) PKC substrate	polyclonal			Rabbit
Ab1-phospho-(Ser/Thr) PKA substrate	polyclonal			Rabbit
Ab1-plcg1	polyclonal			Rabbit
Ab2-mapk14	monoclonal		IgG	Rabbit
Ab2-plcg1	polyclonal			Rabbit
Ab9-mapk	monoclonal		IgG	Rabbit
Ab13-mapk	polyclonal			Rabbit

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Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP
GAL4FF	EFG	GAL4FF
GFP	EFG	GFP
Tomato	EFG	Tomato

Mapping

Shin et al., 2016 ZDB-PUB-160901-3 PMID:27578780

Vegfa signals through ERK to promote angiogenesis, but not artery differentiation

Shin et al., 2016

ZDB-PUB-160901-3
PMID:27578780