PUBLICATION

Zebrafish nephrosin helps host defence against Escherichia coli infection.

Authors

Di, Q., Lin, Q., Huang, Z., Chi, Y., Chen, X., Zhang, W., Zhang, Y.

ID

ZDB-PUB-170825-10

Date

2017

Source

Open Biology 7(8): (Journal)

Registered Authors

Chen, Xiaohui, Chi, Yali, Di, Qianqian, Huang, Zhibin, Lin, Qing, Zhang, Wenqing, Zhang, Yiyue

Keywords

infection, nephrosin, neutrophil, zebrafish

MeSH Terms

Zebrafish Proteins/genetics*
Zebrafish Proteins/metabolism
Embryo, Nonmammalian/immunology
Embryo, Nonmammalian/metabolism
Embryo, Nonmammalian/microbiology
Zebrafish
Animals
Escherichia coli Infections/genetics
Escherichia coli Infections/immunology
Escherichia coli Infections/prevention & control*
CRISPR-Cas Systems
Metalloendopeptidases/genetics*
Metalloendopeptidases/metabolism
Mutation
Gene Knockdown Techniques
Disease Models, Animal
Neutrophils/metabolism*

(all 17)

PubMed

28835569 Full text @ Open Biol.

Abstract

Neutrophils play important roles in innate immunity and are mainly dependent on various enzyme-containing granules to kill engulfed microorganisms. Zebrafish nephrosin (npsn) is specifically expressed in neutrophils; however, its function is largely unknown. Here, we generated an npsn mutant (npsn^smu5 ) via CRISPR/Cas9 to investigate the in vivo function of Npsn. The overall development and number of neutrophils remained unchanged in npsn-deficient mutants, whereas neutrophil antibacterial function was defective. Upon infection with Escherichia coli, the npsn^smu5 mutants exhibited a lower survival rate and more severe bacterial burden, as well as augmented inflammatory response to challenge with infection when compared with wild-type embryos, whereas npsn-overexpressing zebrafish exhibited enhanced host defence against E. coli infection. These findings demonstrated that zebrafish Npsn promotes host defence against bacterial infection. Furthermore, our findings suggested that npsn-deficient and -overexpressing zebrafish might serve as effective models of in vivo innate immunity.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
cxcl8a	npsn^sm15/sm15	bacterial treatment by injection: Escherichia coli	Long-pec	whole organism	RTPCR	Fig. 4
cxcl8a	npsn^sm15/sm15	bacterial treatment by injection: Escherichia coli	Long-pec	whole organism	RTPCR	Fig. 6
cxcl8a	npsn^sm15/sm15; sm13Tg	bacterial treatment by injection: Escherichia coli, heat exposure	Long-pec	whole organism	RTPCR	Fig. 6
cxcl8a	npsn^sm15/+	bacterial treatment by injection: Escherichia coli	Long-pec	whole organism	RTPCR	Fig. 4
cxcl8a	sm13Tg	bacterial treatment by injection: Escherichia coli, heat exposure	Long-pec	whole organism	RTPCR	Fig. 6
GFP	i114Tg	control	Protruding-mouth	neutrophil	IHC	Fig. 1
GFP	i114Tg	bacterial treatment by injection: Escherichia coli	Long-pec	neutrophil	IHC	Fig. 7
GFP	i114Tg	bacterial treatment by injection: Escherichia coli	Protruding-mouth	neutrophil	IHC	Fig. 7
GFP	i114Tg	bacterial treatment by injection: Escherichia coli	Long-pec	neutrophil	IHC	Fig. 5
GFP	i114Tg; sm13Tg	bacterial treatment by injection: Escherichia coli, heat exposure	Protruding-mouth	neutrophil	IHC	Fig. 7

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	bacterial treatment by injection: Escherichia coli	Long-pec to Day 5	whole organism decreased life span, abnormal	Fig. 4
WT	bacterial treatment by injection: Escherichia coli	Long-pec to Days 7-13	whole organism decreased life span, abnormal	Fig. 6
i114Tg; sm13Tg	bacterial treatment by injection: Escherichia coli, heat exposure	Long-pec	defense response to bacterium increased rate, abnormal	Fig. 7
i114Tg; sm13Tg	bacterial treatment by injection: Escherichia coli, heat exposure	Long-pec	neutrophil chemotaxis decreased rate, abnormal	Fig. 7
i114Tg; sm13Tg	bacterial treatment by injection: Escherichia coli, heat exposure	Protruding-mouth	defense response to bacterium increased rate, abnormal	Fig. 7
i114Tg; sm13Tg	bacterial treatment by injection: Escherichia coli, heat exposure	Protruding-mouth	neutrophil chemotaxis decreased rate, abnormal	Fig. 7
npsn^sm15/+	control	Protruding-mouth	whole organism npsn expression decreased amount, abnormal	Fig. 2
npsn^sm15/+	bacterial treatment by injection: Escherichia coli	Long-pec	whole organism cxcl8a expression increased amount, abnormal	Fig. 4
npsn^sm15/+	bacterial treatment by injection: Escherichia coli	Long-pec	whole organism il1b expression increased amount, abnormal	Fig. 4
npsn^sm15/+	bacterial treatment by injection: Escherichia coli	Long-pec	whole organism tnfa expression increased amount, abnormal	Fig. 4

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
gl22Tg	Tg(mpeg1:EGFP)	Transgenic Insertion
i114Tg	Tg(mpx:GFP)	Transgenic Insertion
nz50Tg	Tg(lyz:DsRed2)	Transgenic Insertion
sm12Tg	Tg(npsn:EGFP)	Transgenic Insertion
sm13Tg	Tg(hsp70l:MYC-npsn)	Transgenic Insertion
sm15		Small Deletion	npsn
sm16		Insertion	npsn

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
npsn	CRISPR1-npsn	CRISPR

Fish

Fish
gl22Tg
i114Tg
i114Tg; sm13Tg
npsn^sm15/sm15
npsn^sm15/sm15
npsn^sm15/sm15; i114Tg
npsn^sm15/sm15; sm13Tg
npsn^sm15/+
sm13Tg
WT

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
DsRed2	EFG	DsRed2
EGFP	EFG	EGFP
GFP	EFG	GFP

Mapping

Di et al., 2017 ZDB-PUB-170825-10 PMID:28835569

Zebrafish nephrosin helps host defence against Escherichia coli infection.

Di et al., 2017

ZDB-PUB-170825-10
PMID:28835569