PUBLICATION

Cooperative contributions of the klf1 and klf17 genes in zebrafish primitive erythropoiesis

Authors

Suzuki, H., Ogawa, T., Fujita, S., Sone, R., Kawahara, A.

ID

ZDB-PUB-230811-45

Date

2023

Source

Scientific Reports 13: 1227912279 (Journal)

Registered Authors

Kawahara, Atsuo

Keywords

none

MeSH Terms

Kruppel-Like Transcription Factors/metabolism
Transcription Factors/genetics
Zebrafish*/genetics
Zebrafish*/metabolism
Embryonic Development
Animals
Erythropoiesis*/genetics

(all 7)

PubMed

37563131 Full text @ Sci. Rep.

Abstract

Krüppel-like transcription factors (Klfs), which are characterized by the three conserved C-terminal zinc fingers, are involved in various biological processes, such as haematopoiesis and angiogenesis. However, how the Klf family of transcription factors cooperate in organogenesis remains elusive. During zebrafish embryogenesis, both klf1 and klf17 are expressed in the intermediate cell mass (ICM), where primitive erythroid cells are produced. Using CRISPR-Cas9 genome editing technology, we established klf1-klf17 double mutant zebrafish to investigate the functionally interactive roles of the klf1 and klf17 genes. The klf1-klf17 mutant exhibited a diminished number of circulating primitive erythroid cells at 2 days postfertilization (dpf), while klf1 or klf17 single mutants and wild-type embryos produced comparable numbers of primitive erythroid cells. Circulating erythroid cells from the klf1-klf17 mutant possessed larger nuclei at 2 dpf than wild-type cells, suggesting the impairment of primitive erythroid cell maturation. The expression of the erythroid cell maturation markers band3 and mitoferrin, but not the haematopoietic progenitor markers c-myb and scl, was decreased in the klf1-klf17 mutant at 1 dpf. Thus, these results illustrate the cooperative function of klf1 and klf17 in the maturation processes of zebrafish primitive erythroid cells.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
alas2	klf1^uy19/uy19; klf17^uy21/uy21	standard conditions	Prim-5	intermediate cell mass of mesoderm	ISH	Figure 5
alas2	klf1^uy19/uy19; klf17^uy21/uy21	standard conditions	Prim-5	whole organism	RTPCR	Figure 6
alas2	WT	standard conditions	Prim-5	intermediate cell mass of mesoderm	ISH	Figure 5
alas2	WT	standard conditions	Prim-5	whole organism	RTPCR	Figure 6
EGFP	klf17^uy21/uy21; ko02Tg	standard conditions	26+ somites	yolk common myeloid progenitor	IFL	Figure 2
EGFP	klf1^uy19/uy19; klf17^uy21/uy21; ko02Tg	standard conditions	26+ somites	yolk common myeloid progenitor	IFL	Figure 2
EGFP	klf1^uy19/uy19; ko02Tg	standard conditions	26+ somites	yolk common myeloid progenitor	IFL	Figure 2
EGFP	ko02Tg	standard conditions	26+ somites	yolk common myeloid progenitor	IFL	Figure 2
fech	klf1^uy19/uy19; klf17^uy21/uy21	standard conditions	Prim-5	intermediate cell mass of mesoderm	ISH	Figure 5
fech	klf1^uy19/uy19; klf17^uy21/uy21	standard conditions	Prim-5	whole organism	RTPCR	Figure 6

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
klf17^uy21/uy21	standard conditions	Long-pec	erythroblast normal amount, normal	Figure 1
klf17^uy21/uy21; ko02Tg	standard conditions	26+ somites	yolk common myeloid progenitor normal amount, normal	Figure 2
klf1^uy19/uy19	standard conditions	Long-pec	erythroblast normal amount, normal	Figure 1
klf1^uy19/uy19; klf17^uy21/uy21	standard conditions	Prim-5	intermediate cell mass of mesoderm alas2 expression decreased distribution, abnormal	Figure 5
klf1^uy19/uy19; klf17^uy21/uy21	standard conditions	Prim-5	intermediate cell mass of mesoderm slc25a37 expression decreased distribution, abnormal	Figure 5
klf1^uy19/uy19; klf17^uy21/uy21	standard conditions	Prim-5	intermediate cell mass of mesoderm slc4a1a expression decreased distribution, abnormal	Figure 5
klf1^uy19/uy19; klf17^uy21/uy21	standard conditions	Prim-5	whole organism alas2 expression decreased amount, abnormal	Figure 6
klf1^uy19/uy19; klf17^uy21/uy21	standard conditions	Prim-5	whole organism slc25a37 expression decreased amount, abnormal	Figure 6
klf1^uy19/uy19; klf17^uy21/uy21	standard conditions	Prim-5	whole organism slc4a1a expression decreased amount, abnormal	Figure 6
klf1^uy19/uy19; klf17^uy21/uy21	standard conditions	Long-pec	erythroblast decreased amount, abnormal	Figure 1

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
ko02Tg	Tg(-2.4lyz:EGFP)	Transgenic Insertion
ko05Tg	Tg(-8.1gata1a:mRFP)	Transgenic Insertion
uy19		Small Deletion	klf1
uy21		Indel	klf17

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
klf1	CRISPR1-klf1	CRISPR
klf1	CRISPR2-klf1	CRISPR

Fish

Fish
klf1^uy19/uy19
klf1^uy19/uy19; klf17^uy21/uy21
klf1^uy19/uy19; klf17^uy21/uy21; ko02Tg
klf1^uy19/uy19; klf17^uy21/uy21; ko05Tg
klf1^uy19/uy19; ko02Tg
klf17^uy21/uy21
klf17^uy21/uy21; ko02Tg
ko02Tg
ko05Tg
WT

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Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP
mRFP	EFG	mRFP

Mapping

Suzuki et al., 2023 ZDB-PUB-230811-45 PMID:37563131

Cooperative contributions of the klf1 and klf17 genes in zebrafish primitive erythropoiesis

Suzuki et al., 2023

ZDB-PUB-230811-45
PMID:37563131