PUBLICATION

Comparative analysis of zebrafish nos2a and nos2b genes

Authors

Lepiller, S., Franche, N., Solary, E., Chluba, J., and Laurens, V.

ID

ZDB-PUB-090616-32

Date

2009

Source

Gene 445(1-2): 58-65 (Journal)

Registered Authors

Keywords

inducible nitric oxide synthase, gene duplication, synteny, evolution, teleostei

MeSH Terms

Zebrafish/genetics*
Sequence Homology, Amino Acid
Gene Expression Regulation, Developmental
Phylogeny
Isoenzymes/genetics
Nitric Oxide Synthase Type II/genetics*
Synteny
Animals
Cells, Cultured
Gene Duplication
Molecular Sequence Data
Gene Expression Regulation, Enzymologic
Embryo, Nonmammalian
Cloning, Molecular
Amino Acid Sequence

(all 15)

PubMed

19501636 Full text @ Gene

Abstract

Nitric oxide synthase (NOS) produces nitric oxide (NO) from arginine. Three NOS isoforms have been identified in mammals, namely a neuronal (NOS1), an inducible (NOS2) and an endothelial (NOS3) enzyme. In zebrafish genome, one nos1 gene and two nos2 genes (nos2a and nos2b) were observed. We cloned zebrafish nos2a cDNA and compared nos2a and nos2b sequences, expression and inducibility. When analyzed by reverse transcription-PCR, the expression of nos2a remained very low during initial development, then increased at 96hpf, while nos2b was expressed from 6hpf and subsequently remained stable. Expression of nos2a is detected in head, eye and gut regions by WISH experiments performed on 48, 72 and 96hpf larvae. In adults, nos2a expression varies from one tissue to another whereas nos2b is expressed in all studied tissues. Both nos2 isoforms can be induced by pro-inflammatory or mechanical stresses (tissue injury). In vitro as in vivo stimulations with Poly I:C and lipopolysaccharides (LPS) enhanced more dramatically nos2a than nos2b expression. After tail transection in 4dpf larvae a strong increase of nos2a and nos2b expression was evidenced in the regeneration site, skin cells and for the nos2b gene in neuromasts. Phylogenetic and syntenic analyses show that nos2b gene was associated with syntenic genes identified for nos2 genes in vertebrate. This is not the case for the nos2a gene, despite zebrafish nos2a presents the inducible property of a classical vertebrate nos2 isoform. A myristoylation consensus site was detected at the N-terminal extremity of zebrafish Nos2b, a property shared with mammals NOS3 isoforms. Thus, the evolution of nos2 genes in zebrafish provides a typical example of gene divergence after duplication.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Qualifier	Anatomy	Assay	Figure
ifnphi1	WT	chemical treatment: lipopolysaccharide	Adult		whole organism	RTPCR	Fig. 7
ifnphi1	WT	control	Adult	Not Detected	whole organism	RTPCR	Fig. 7
nos2a	WT	standard conditions	Adult		brain	RTPCR	Fig. 6
nos2a	WT	standard conditions	Adult		eye	RTPCR	Fig. 6
nos2a	WT	standard conditions	Long-pec to Protruding-mouth		eye	ISH	Fig. 5
nos2a	WT	standard conditions	Day 4		eye	ISH	Fig. 5
nos2a	WT	standard conditions	Adult		gill	RTPCR	Fig. 6
nos2a	WT	standard conditions	Day 4		gut	ISH	Fig. 5
nos2a	WT	standard conditions	Adult		gut	RTPCR	Fig. 6
nos2a	WT	standard conditions	Day 4		head	ISH	Fig. 5

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Phenotype

Mutations / Transgenics

Human Disease / Model

Sequence Targeting Reagents

Fish

Fish
WT

Antibodies

Orthology

Gene	Orthology
nos1
nos2a
nos2b

Engineered Foreign Genes

Mapping

Lepiller et al., 2009 ZDB-PUB-090616-32 PMID:19501636

Comparative analysis of zebrafish nos2a and nos2b genes

Lepiller et al., 2009

ZDB-PUB-090616-32
PMID:19501636