PUBLICATION

Wnt9a Is Required for the Aortic Amplification of Nascent Hematopoietic Stem Cells

Authors

Grainger, S., Richter, J., Palazón, R.E., Pouget, C., Lonquich, B., Wirth, S., Grassme, K.S., Herzog, W., Swift, M.R., Weinstein, B.M., Traver, D., Willert, K.

ID

ZDB-PUB-161103-1

Date

2016

Source

Cell Reports 17: 1595-1606 (Journal)

Registered Authors

Grainger, Stephanie, Grassme, Kathrin, Herzog, Wiebke, Swift, Matthew Russell, Traver, David, Weinstein, Brant M.

Keywords

HSCs, HSPCs, Myc, Wnt, Wnt9a, aorta, hematopoietic stem and progenitor cells, hematopoietic stem cells, proliferation

MeSH Terms

Animals
Hemangioblasts/metabolism
Cell Cycle
Hematopoietic Stem Cells/cytology*
Hematopoietic Stem Cells/metabolism*
Cell Count
Zebrafish Proteins/metabolism*
Wnt Proteins/metabolism*
Wnt Signaling Pathway
Cell Proliferation
Aorta/cytology*
Aorta/embryology*
Zebrafish/embryology*

(all 13)

PubMed

27806298 Full text @ Cell Rep.

Abstract

All mature blood cell types in the adult animal arise from hematopoietic stem and progenitor cells (HSPCs). However, the developmental cues regulating HSPC ontogeny are incompletely understood. In particular, the details surrounding a requirement for Wnt/β-catenin signaling in the development of mature HSPCs are controversial and difficult to consolidate. Using zebrafish, we demonstrate that Wnt signaling is required to direct an amplification of HSPCs in the aorta. Wnt9a is specifically required for this process and cannot be replaced by Wnt9b or Wnt3a. This proliferative event occurs independently of initial HSPC fate specification, and the Wnt9a input is required prior to aorta formation. HSPC arterial amplification occurs prior to seeding of secondary hematopoietic tissues and proceeds, in part, through the cell cycle regulator myca (c-myc). Our results support a general paradigm, in which early signaling events, including Wnt, direct later HSPC developmental processes.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
ccnd1	w26Tg; y171Tg	standard conditions	Prim-15	endothelial cell	RTPCR	Fig. 5
ccnd2b	w26Tg; y171Tg	standard conditions	Prim-15	endothelial cell	RTPCR	Fig. 5
cdk2	w26Tg; y171Tg	standard conditions	Prim-15	endothelial cell	RTPCR	Fig. 5
cdk4	w26Tg; y171Tg	standard conditions	Prim-15	endothelial cell	RTPCR	Fig. 5
EGFP	y171Tg; zf169Tg	chemical treatment by environment: lithium chloride	Prim-25	ventral wall of dorsal aorta hematopoietic stem cell	IFL	Fig. 1
EGFP	y171Tg; zf169Tg	control	Prim-25	ventral wall of dorsal aorta hematopoietic stem cell	IFL	Fig. 1
EGFP	y171Tg; zf169Tg	control	Prim-25	ventral wall of dorsal aorta hematopoietic stem cell	IFL	Fig. 3
EGFP	y171Tg; zf169Tg	chemical treatment by environment: Wnt signalling inhibitor	Prim-25	ventral wall of dorsal aorta hematopoietic stem cell	IFL	Fig. 1
EGFP	y171Tg; zf169Tg + MO1-wnt9a	standard conditions	Prim-25	ventral wall of dorsal aorta hematopoietic stem cell	IFL	Fig. 3
EGFP	y171Tg; zf169Tg + MO1-wnt9b	standard conditions	Prim-25	ventral wall of dorsal aorta hematopoietic stem cell	IFL	Fig. 3

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
AB + MO2-wnt9a	standard conditions	Prim-25	ventral wall of dorsal aorta hematopoietic stem cell myb expression decreased amount, abnormal	Fig. 5
AB + MO2-wnt9a	standard conditions	Prim-25	ventral wall of dorsal aorta hematopoietic stem cell decreased amount, abnormal	Fig. 5
AB + MO2-wnt9a	standard conditions	Day 4	ventral wall of dorsal aorta hematopoietic stem cell myb expression decreased amount, abnormal	Fig. 3
AB + MO2-wnt9a	standard conditions	Day 4	ventral wall of dorsal aorta hematopoietic stem cell decreased amount, abnormal	Fig. 3
mu271Tg; sd32Tg; y171Tg	chemical treatment by environment: 5-fluorouracil	Prim-25	ventral wall of dorsal aorta hematopoietic stem cell decreased amount, abnormal	Fig. 4
s843Tg; w141Tg + MO2-wnt9a	standard conditions	Prim-5	endothelial cell gata2b expression increased amount, abnormal	Fig. 4
s843Tg; w141Tg + MO2-wnt9a	standard conditions	Prim-5	hematopoietic stem cell mitotic G1 phase arrested, abnormal	Fig. 4
s843Tg; w141Tg + MO2-wnt9a	standard conditions	Prim-5	whole organism gata2b expression increased amount, abnormal	Fig. 4
sd32Tg; sd46Tg	heat shock	Prim-25	ventral wall of dorsal aorta hematopoietic stem cell myb expression decreased amount, abnormal	Fig. 2
sd32Tg; sd46Tg	heat shock	Prim-25	ventral wall of dorsal aorta hematopoietic stem cell decreased amount, abnormal	Fig. 2

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
hzm7Et	Et(shhb:KALTA4,UAS-E1B:mCherry)	Transgenic Insertion
ia4Tg	Tg(7xTCF-Xla.Sia:GFP)	Transgenic Insertion
kca4Tg	Tg(-1.5hsp70l:GAL4)	Transgenic Insertion
mu271Tg	Tg(UAS:LIFEACT-GFP)	Transgenic Insertion
s843Tg	Tg(kdrl:EGFP)	Transgenic Insertion
sd32Tg	TgBAC(gata2b:KALTA4)	Transgenic Insertion
sd46Tg	Tg(UAS:dntcf7l1a-2A-mKate2,myl7:EGFP)	Transgenic Insertion
sd47Tg	Tg(UAS:cactnnb1-2A-mKate2,myl7:EGFP)	Transgenic Insertion
sd49		Small Deletion	wnt9a
sd50Tg	Tg(UAS:wnt9a-IRES-TagBFP)	Transgenic Insertion

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
wnt9a	CRISPR1-wnt9a	CRISPR
wnt9a	MO1-wnt9a	MRPHLNO
wnt9a	MO2-wnt9a	MRPHLNO
wnt9b	MO1-wnt9b	MRPHLNO

Fish

Fish
AB
AB + MO2-wnt9a
mu271Tg; sd32Tg
mu271Tg; sd32Tg; y171Tg
s843Tg; w141Tg
s843Tg; w141Tg + MO2-wnt9a
sd32Tg; sd46Tg
sd46Tg; ubs4Tg
w26Tg
w26Tg; y171Tg

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
Cerulean	EFG	Cerulean
EGFP	EFG	EGFP
GAL4	EFG	GAL4
GAL4FF	EFG	GAL4FF
GFP	EFG	GFP
KALTA4	EFG	KALTA4
mCherry	EFG	mCherry
mKate2	EFG	mKate2
TagBFP	EFG	TagBFP

Mapping

Grainger et al., 2016 ZDB-PUB-161103-1 PMID:27806298

Wnt9a Is Required for the Aortic Amplification of Nascent Hematopoietic Stem Cells

Grainger et al., 2016

ZDB-PUB-161103-1
PMID:27806298