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PMID:19501636

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Citation

Lepiller, S, Franche, N, Solary, E, Chluba, J and Laurens, V (2009) Comparative analysis of zebrafish nos2a and nos2b genes. Gene 445:58-65

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 96 hpf, while nos2b was expressed from 6 hpf and subsequently remained stable. Expression of nos2a is detected in the head, eye and gut regions by WISH experiments performed at 48, 72 and 96 hpf 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 4 dpf 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 presenting 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 mammal NOS3 isoforms. Thus, the evolution of nos2 genes in zebrafish provides a typical example of gene divergence after duplication.

Links

PubMed Online version:10.1016/j.gene.2009.05.016

Keywords

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

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

DANRE:A2BIN8

involved_in

GO:0032496: response to lipopolysaccharide

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DANRE:A2BIN8

involved_in

GO:0009611: response to wounding

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DANRE:A7WK81

involved_in

GO:0032496: response to lipopolysaccharide

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DANRE:A7WK81

involved_in

GO:0009611: response to wounding

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DANRE:A9JNR5

involved_in

GO:0032496: response to lipopolysaccharide

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DANRE:A9JNR5

involved_in

GO:0009611: response to wounding

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DANRE:B0FL42

involved_in

GO:0032496: response to lipopolysaccharide

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DANRE:B0FL42

involved_in

GO:0009611: response to wounding

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DANRE:F1QML0

involved_in

GO:0032496: response to lipopolysaccharide

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DANRE:F1QML0

involved_in

GO:0009611: response to wounding

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DANRE:Q7T1G1

involved_in

GO:0032496: response to lipopolysaccharide

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DANRE:Q7T1G1

involved_in

GO:0009611: response to wounding

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete


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References

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