PUBLICATION

The ETS transcription factor Spi2 regulates hematopoietic cell development in zebrafish

Authors

Zhao, S., Zhang, A., Zhu, H., Wen, Z.

ID

ZDB-PUB-221001-3

Date

2022

Source

Development (Cambridge, England) 149(19): (Journal)

Registered Authors

Wen, Zilong

Keywords

ETS transcription factors, Hematopoiesis, Hematopoietic stem/progenitor cells, Zebrafish

MeSH Terms

Animals
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism
Hemangioblasts*/metabolism
Core Binding Factor Alpha 2 Subunit/genetics
Core Binding Factor Alpha 2 Subunit/metabolism
Hematopoiesis/genetics
Proto-Oncogene Proteins c-ets/genetics
Proto-Oncogene Proteins c-ets/metabolism
Zebrafish*/genetics
Zebrafish*/metabolism

(all 11)

PubMed

36178053 Full text @ Development

Abstract

The E26 transformation-specific or E-twenty-six (ETS) genes encode a superfamily of transcription factors involved in diverse biological processes. Here, we report the identification and characterization of a previously unidentified member of the ETS transcription factors, Spi2, that is found exclusively in the ray-finned fish kingdom. We show that the expression of spi2 is restricted to hemogenic endothelial cells (HECs) and to hematopoietic stem and progenitor cells (HSPCs) in zebrafish. Using bacteria artificial chromosome transgenesis, we generate a spi2 reporter line, TgBAC(spi2:P2a-GFP), which manifests the GFP pattern recapitulating the endogenous spi2 expression. Genetic ablation of spi2 has little effect on HEC formation and the endothelial-to-hematopoietic transition, but results in compromised proliferation of HSPCs in the caudal hematopoietic tissue (CHT) during early development and in severe myeloid lineage defect in adulthood. Epistatic analysis shows that spi2 acts downstream of runx1 in regulating HSPC development in the CHT. Our study identifies Spi2 as an essential regulator for definitive hematopoietic cell development and creates a TgBAC(spi2:P2a-GFP) reporter line for tracking HECs, HSPCs, myeloid cells and thrombocytes from early development to adulthood.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
DsRed	spi2^hkz58/hkz58; hkz015tTg/hkz015tTg	standard conditions	Days 14-20	macrophage	IFL	Fig. 3
DsRed	spi2^hkz58/hkz58; nz50Tg/nz50Tg	standard conditions	Days 14-20	ventral wall of dorsal aorta neutrophil	IFL	Fig. 3
EGFP	spi2^hkz58/hkz58; s843Tg/s843Tg	standard conditions	Prim-15	dorsal aorta	IFL	Fig. 3
GFP	hkz012tTg/hkz012tTg; hkz046Tg/hkz046Tg	standard conditions	Protruding-mouth	caudal hematopoietic tissue leukocyte	IFL	Fig. 2
GFP	hkz046Tg/hkz046Tg	standard conditions	Prim-15	dorsal aorta basal side	IFL	Fig. 2
GFP	hkz046Tg/hkz046Tg; sd2Tg/sd2Tg	standard conditions	Protruding-mouth	caudal hematopoietic tissue erythroid progenitor cell	IFL	Fig. 2
hbae1.1	spi2^hkz58/hkz58	standard conditions	Day 5	nucleate erythrocyte	ISH	Fig. 3
hbae1.1	spi2^hkz58/+	standard conditions	Day 5	nucleate erythrocyte	ISH	Fig. 3
myb	spi2^hkz58/hkz58	standard conditions	Day 5	caudal hematopoietic tissue	ISH	Fig. 3
myb	spi2^hkz58/hkz58	standard conditions	Long-pec	caudal hematopoietic tissue	ISH	Fig. 3

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
hkz041Tg/hkz041Tg	standard conditions	Prim-5	ventral wall of dorsal aorta runx1 expression increased amount, abnormal	Fig. 4
hkz041Tg/hkz041Tg	standard conditions	Prim-5	ventral wall of dorsal aorta spi2 expression increased amount, abnormal	Fig. 4
hkz046Tg/hkz046Tg	standard conditions	Adult	head kidney hematopoietic multipotent progenitor cell increased amount, normal	Fig. 2
hkz046Tg/hkz046Tg	standard conditions	Adult	head kidney myeloid cell increased amount, normal	Fig. 2
runx1^hg1/hg1	standard conditions	Prim-5	ventral wall of dorsal aorta spi2 expression decreased amount, abnormal	Fig. 4
runx1^hg1/hg1	standard conditions	Long-pec	caudal hematopoietic tissue spi2 expression decreased amount, abnormal	Fig. 4
runx1^hg1/hg1	standard conditions	Long-pec	caudal hematopoietic tissue spi2 expression decreased distribution, abnormal	Fig. 4
spi2^hkz58/hkz58	standard conditions	Long-pec	caudal hematopoietic tissue myb expression decreased distribution, abnormal	Fig. 3
spi2^hkz58/hkz58	standard conditions	Protruding-mouth	caudal hematopoietic tissue myb expression decreased amount, abnormal	Fig. 4
spi2^hkz58/hkz58	standard conditions	Protruding-mouth	caudal hematopoietic tissue myb expression decreased distribution, abnormal	Fig. 3

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
cz3325Tg	Tg(LCR:EGFP)	Transgenic Insertion
hg1		Point Mutation	runx1
hkz012tTg	Tg(coro1a:LOXP-DsRedx-LOXP-GFP)	Transgenic Insertion
hkz015tTg	Tg(mpeg1.1:LOXP-DsRedx-LOXP-EGFP)	Transgenic Insertion
hkz016tTg	Tg(lck:LOXP-DsRedx-LOXP-EGFP)	Transgenic Insertion
hkz041Tg	Tg(kdrl:runx1-2A-GFP)	Transgenic Insertion
hkz046Tg	TgBAC(spi2:spi2-2A-GFP)	Transgenic Insertion
hkz58		Small Deletion	spi2
la2Tg	Tg(-6.0itga2b:EGFP)	Transgenic Insertion
ncv6Tg	Tg(kdrl:NLS-Eos)	Transgenic Insertion

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
spi2	CRISPR1-spi2	CRISPR
spi2	CRISPR2-spi2	CRISPR

Fish

Fish
AB
hkz012tTg/hkz012tTg; hkz046Tg/hkz046Tg
hkz041Tg/hkz041Tg
hkz046Tg/hkz046Tg
hkz046Tg/hkz046Tg; sd2Tg/sd2Tg
la2Tg/la2Tg
ncv6Tg/ncv6Tg
runx1^hg1/hg1
s843Tg/s843Tg
spi2^hkz58/hkz58

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
DsRed	EFG	DsRed
DsRed2	EFG	DsRed2
DsRedx	EFG	DsRedx
EGFP	EFG	EGFP
Eos	EFG	Eos
GFP	EFG	GFP

Mapping

Zhao et al., 2022 ZDB-PUB-221001-3 PMID:36178053

The ETS transcription factor Spi2 regulates hematopoietic cell development in zebrafish

Zhao et al., 2022

ZDB-PUB-221001-3
PMID:36178053