PUBLICATION

Blood stem cell-forming haemogenic endothelium in zebrafish derives from arterial endothelium

Authors

Bonkhofer, F., Rispoli, R., Pinheiro, P., Krecsmarik, M., Schneider-Swales, J., Tsang, I.H.C., de Bruijn, M., Monteiro, R., Peterkin, T., Patient, R.

ID

ZDB-PUB-190810-3

Date

2019

Source

Nature communications 10: 3577 (Journal)

Registered Authors

Monteiro, Rui, Patient, Roger K., Peterkin, Tessa, Pinheiro, Philip

Keywords

none

Datasets

GEO:GSE132259, GEO:GSE132258, GEO:GSE132260

MeSH Terms

Animals
Gene Knockout Techniques
Zebrafish/embryology*
Zebrafish/metabolism
Cell Lineage
Intracellular Signaling Peptides and Proteins/metabolism
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism
Embryonic Development
Arteries/cytology
Arteries/embryology*
Arteries/metabolism
Animals, Genetically Modified
Endothelium, Vascular/cytology
Endothelium, Vascular/embryology*
Endothelium, Vascular/metabolism
Hemangioblasts/physiology*
Core Binding Factor Alpha 2 Subunit/genetics
Embryo, Nonmammalian

(all 19)

PubMed

31395869 Full text @ Nat. Commun.

Abstract

Haematopoietic stem cells are generated from the haemogenic endothelium (HE) located in the floor of the dorsal aorta (DA). Despite being integral to arteries, it is controversial whether HE and arterial endothelium share a common lineage. Here, we present a transgenic zebrafish runx1 reporter line to isolate HE and aortic roof endothelium (ARE)s, excluding non-aortic endothelium. Transcriptomic analysis of these populations identifies Runx1-regulated genes and shows that HE initially expresses arterial markers at similar levels to ARE. Furthermore, runx1 expression depends on prior arterial programming by the Notch ligand dll4. Runx1^-/- mutants fail to downregulate arterial genes in the HE, which remains integrated within the DA, suggesting that Runx1 represses the pre-existing arterial programme in HE to allow progression towards the haematopoietic fate. These findings strongly suggest that, in zebrafish, aortic endothelium is a precursor to HE, with potential implications for pluripotent stem cell differentiation protocols for the generation of transplantable HSCs.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Qualifier	Anatomy	Assay	Figure
angpt1	runx1^hg1/hg1	standard conditions	Prim-5		ventral wall of dorsal aorta	ISH	Fig. 4
angpt1	WT	standard conditions	Prim-5		ventral wall of dorsal aorta	ISH	Fig. 4
Citrine	ox1Tg	standard conditions	Day 6		caudal hematopoietic tissue	IFL	Fig. S1
Citrine	ox1Tg	standard conditions	Day 6		kidney	IFL	Fig. S1
Citrine	ox1Tg	standard conditions	1-4 somites		lateral plate mesoderm	ISH	Fig. S1
Citrine	ox1Tg	standard conditions	5-9 somites		posterior lateral plate mesoderm	IFL	Fig. S1
Citrine	ox1Tg	standard conditions	14-19 somites		posterior lateral plate mesoderm	IFL	Fig. S1
Citrine	ox1Tg	standard conditions	5-9 somites		posterior lateral plate mesoderm	ISH	Fig. S1
Citrine	ox1Tg	standard conditions	10-13 somites		posterior lateral plate mesoderm	ISH	Fig. 1
Citrine	ox1Tg	standard conditions	10-13 somites	Not Detected	Rohon-Beard neuron	ISH	Fig. 1

1 - 10 of 55

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT + MO1-cldn5b	standard conditions	Prim-5	ventral wall of dorsal aorta dll4 expression increased amount, abnormal	Fig. S9
WT + MO1-cldn5b	standard conditions	Prim-5	ventral wall of dorsal aorta runx1 expression decreased amount, abnormal	Fig. S9
WT + MO1-dll4	standard conditions	Prim-15	ventral wall of dorsal aorta runx1 expression decreased amount, abnormal	Fig. 6
WT + MO4-dll4	standard conditions	Prim-15	ventral wall of dorsal aorta runx1 expression decreased amount, abnormal	Fig. 6
dll4^{sa9436/sa9436}	standard conditions	Prim-15	ventral wall of dorsal aorta runx1 expression decreased amount, abnormal	Fig. 6
ox1Tg + MO1-gata2b	standard conditions	Prim-15	ventral wall of dorsal aorta Citrine expression decreased amount, abnormal	Fig. S1
ox1Tg + MO8-tal1	standard conditions	Prim-15	ventral wall of dorsal aorta Citrine expression decreased amount, abnormal	Fig. S1
runx1^hg1/hg1	standard conditions	Prim-5	ventral wall of dorsal aorta angpt1 expression decreased amount, abnormal	Fig. 4
runx1^hg1/hg1	standard conditions	Prim-5	ventral wall of dorsal aorta dll4 expression increased amount, abnormal	Fig. 5
runx1^hg1/hg1	standard conditions	Prim-5	ventral wall of dorsal aorta sox17 expression increased amount, abnormal	Fig. 5

1 - 10 of 17

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
hg1		Point Mutation	runx1
ox1Tg	TgBAC(runx1:Citrine)	Transgenic Insertion
s896Tg	Tg(kdrl:Hsa.HRAS-mCherry)	Transgenic Insertion
sa9436		Point Mutation	dll4

1 - 4 of 4

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
cldn5b	MO1-cldn5b	MRPHLNO
dll4	MO1-dll4	MRPHLNO
dll4	MO4-dll4	MRPHLNO
gata2b	MO1-gata2b	MRPHLNO
runx1	MO1-runx1	MRPHLNO
tal1	MO8-tal1	MRPHLNO

Fish

Fish
dll4^{sa9436/sa9436}
ox1Tg
ox1Tg + MO1-gata2b
ox1Tg + MO8-tal1
ox1Tg; s896Tg
runx1^hg1/hg1
runx1^hg1/hg1; ox1Tg; s896Tg
WT
WT + MO1-cldn5b
WT + MO1-dll4

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
Citrine	EFG	Citrine
mCherry	EFG	mCherry

Mapping

Bonkhofer et al., 2019 ZDB-PUB-190810-3 PMID:31395869

Blood stem cell-forming haemogenic endothelium in zebrafish derives from arterial endothelium

Bonkhofer et al., 2019

ZDB-PUB-190810-3
PMID:31395869