PUBLICATION

Single-cell transcriptomic analysis identifies the conversion of zebrafish Etv2-deficient vascular progenitors into skeletal muscle

Authors

Chestnut, B., Casie Chetty, S., Koenig, A.L., Sumanas, S.

ID

ZDB-PUB-200606-5

Date

2020

Source

Nature communications 11: 2796 (Journal)

Registered Authors

Sumanas, Saulius

Keywords

none

Datasets

GEO:GSE142484

MeSH Terms

Wnt Signaling Pathway
Somites/metabolism
Models, Biological
Single-Cell Analysis*
Embryo, Nonmammalian/metabolism
Heat-Shock Response
Blood Vessels/cytology*
Mutation/genetics
Zebrafish/embryology
Zebrafish/genetics*
Cell Differentiation/genetics
Gene Expression Profiling*
Zebrafish Proteins/deficiency*
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism
Green Fluorescent Proteins/metabolism
Stem Cells/metabolism*
Cell Movement
Animals, Genetically Modified
Animals
Transcription, Genetic
Fibroblast Growth Factors/metabolism
Biomarkers/metabolism
Muscle, Skeletal/cytology*

(all 24)

PubMed

32493965 Full text @ Nat. Commun.

Abstract

Cell fate decisions involved in vascular and hematopoietic embryonic development are still poorly understood. An ETS transcription factor Etv2 functions as an evolutionarily conserved master regulator of vasculogenesis. Here we report a single-cell transcriptomic analysis of hematovascular development in wild-type and etv2 mutant zebrafish embryos. Distinct transcriptional signatures of different types of hematopoietic and vascular progenitors are identified using an etv2^ci32Gt gene trap line, in which the Gal4 transcriptional activator is integrated into the etv2 gene locus. We observe a cell population with a skeletal muscle signature in etv2-deficient embryos. We demonstrate that multiple etv2^ci32Gt; UAS:GFP cells differentiate as skeletal muscle cells instead of contributing to vasculature in etv2-deficient embryos. Wnt and FGF signaling promote the differentiation of these putative multipotent etv2 progenitor cells into skeletal muscle cells. We conclude that etv2 actively represses muscle differentiation in vascular progenitors, thus restricting these cells to a vascular endothelial fate.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
cldn5b	WT	standard conditions	Prim-5	dorsal aorta	ISH	Fig. 8
cldn5b	WT	standard conditions	20-25 somites	vascular cord	ISH	Fig. 8
dab2	WT	standard conditions	Prim-5	posterior cardinal vein	ISH	Fig. 8
dab2	WT	standard conditions	20-25 somites	vascular cord	ISH	Fig. 8
drl	WT	standard conditions	20-25 somites	endocardium	ISH	Fig. 2
EGFP	etsrp^{ci32Gt/ci32Gt}; nkuasgfp1aTg	standard conditions	Prim-5	skeletal muscle cell	IFL	Fig. 3
EGFP	etsrp^{ci32Gt/ci32Gt}; nkuasgfp1aTg + MO1-etsrp + MO2-etsrp	standard conditions	Prim-15	mesoderm	IFL	Fig. 3
EGFP	etsrp^{ci32Gt/ci32Gt}; nkuasgfp1aTg + MO1-etsrp + MO2-etsrp	standard conditions	Prim-15	skeletal muscle cell	IFL	Fig. 3
EGFP	etsrp^ci32Gt/+; c264Tg; nkuasgfp1aTg	standard conditions	Prim-15	skeletal muscle cell	IFL	Fig. 3
EGFP	etsrp^ci32Gt/+; nkuasgfp1aTg	standard conditions	Prim-15	intersegmental vessel	IFL	Fig. 3

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
etsrp^ci32Gt/+; c264Tg; pd1Tg	heat shock	20-25 somites	skeletal muscle cell differentiation decreased occurrence, abnormal	Fig. 6
etsrp^ci32Gt/+; nkuasgfp1aTg	standard conditions	Prim-15	intersegmental vessel EGFP expression decreased amount, abnormal	Fig. 3
etsrp^ci32Gt/+; nkuasgfp1aTg	standard conditions	Prim-15	skeletal muscle cell EGFP expression increased amount, abnormal	Fig. 3
etsrp^ci32Gt/+; nkuasgfp1aTg + MO1-etsrp + MO2-etsrp	standard conditions	Prim-5	skeletal muscle cell differentiation increased occurrence, abnormal	Fig. 3
etsrp^ci32Gt/+; nkuasgfp1aTg; w32Tg	heat shock	Prim-5	skeletal muscle cell differentiation decreased occurrence, abnormal	Fig. 6
etsrp^{ci32Gt/ci32Gt}; nkuasgfp1aTg	standard conditions	10-13 somites	blood vessel endothelial cell differentiation decreased occurrence, abnormal	Fig. 5
etsrp^{ci32Gt/ci32Gt}; nkuasgfp1aTg	standard conditions	10-13 somites	lateral plate mesoderm apoptotic process increased occurrence, abnormal	Fig. 5
etsrp^{ci32Gt/ci32Gt}; nkuasgfp1aTg	standard conditions	10-13 somites	skeletal muscle cell differentiation increased occurrence, abnormal	Fig. 5
etsrp^{ci32Gt/ci32Gt}; nkuasgfp1aTg	standard conditions	14-19 somites	blood vessel endothelial cell differentiation decreased occurrence, abnormal	Fig. 4
etsrp^{ci32Gt/ci32Gt}; nkuasgfp1aTg	standard conditions	14-19 somites	lateral plate mesoderm apoptotic process increased occurrence, abnormal	Fig. 4

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
c264Tg	Tg(UAS-E1B:NTR-mCherry)	Transgenic Insertion
ci1Tg	TgBAC(etsrp:EGFP)	Transgenic Insertion
ci5Tg	Tg(-6.5kdrl:mCherry)	Transgenic Insertion
ci32Gt	Gt(:CRISPR2-etsrp-2A-GAL4)	Transgenic Insertion	etsrp
ci33		Insertion	etsrp
ci46Tg	Tg(5xUAS:LOXP-mCerulean-LOXP-NTR-2A-YFP)	Transgenic Insertion
nkuasgfp1aTg	Tg(5xUAS:EGFP)	Transgenic Insertion
pd1Tg	Tg(hsp70l:dnfgfr1a-EGFP)	Transgenic Insertion
sd2Tg	Tg(gata1a:DsRed)	Transgenic Insertion
w32Tg	Tg(hsp70l:dkk1b-GFP)	Transgenic Insertion

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
etsrp	CRISPR2-etsrp	CRISPR
etsrp	MO1-etsrp	MRPHLNO
etsrp	MO2-etsrp	MRPHLNO
tal1	MO4-tal1	MRPHLNO

Fish

Fish
etsrp^{ci32Gt/ci32Gt}
etsrp^{ci32Gt/ci32Gt}; nkuasgfp1aTg
etsrp^{ci32Gt/ci32Gt}; nkuasgfp1aTg + MO1-etsrp + MO2-etsrp
etsrp^ci32Gt/+
etsrp^ci32Gt/+; c264Tg
etsrp^ci32Gt/+; c264Tg; nkuasgfp1aTg
etsrp^ci32Gt/+; c264Tg; pd1Tg
etsrp^ci32Gt/+; c264Tg; zf13Tg
etsrp^ci32Gt/+; nkuasgfp1aTg
etsrp^ci32Gt/+; nkuasgfp1aTg + MO1-etsrp + MO2-etsrp

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Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab-F310	monoclonal		IgG1	Mouse
Ab-S58	monoclonal		IgA	Mouse

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
Cerulean	EFG	Cerulean
DsRed	EFG	DsRed
EGFP	EFG	EGFP
GAL4	EFG	GAL4
GFP	EFG	GFP
mCherry	EFG	mCherry
NTR	EFG	NTR
YFP	EFG	YFP

Mapping

Chestnut et al., 2020 ZDB-PUB-200606-5 PMID:32493965

Single-cell transcriptomic analysis identifies the conversion of zebrafish Etv2-deficient vascular progenitors into skeletal muscle

Chestnut et al., 2020

ZDB-PUB-200606-5
PMID:32493965