TY - JOUR
T1 - Comparative analysis of zebrafish nos2a and nos2b genes
AU - Lepiller, Sandrine
AU - Franche, Nathalie
AU - Solary, Eric
AU - Chluba, Johanna
AU - Laurens, Véronique
N1 - Funding Information:
We thank Hervé Bouhin (UMR CNRS 5548, Dijon) and Thibault de Malliard (Master studient) for their help in phylogenetic analysis, Muriel Tauzin and Philippe Herbomel (Institut Pasteur, Paris) and Patrick Blader (UMR CNRS 5547, Toulouse) for their advices in whole-mount in situ hybridization. We are grateful to Michael Schorpp (Max Planck Institut, Freiburg) for being so generous with his time and knowledge, welcoming V.L. to learn zebrafish methods in his lab. This work was supported by grants to J.C. from the Ligue Contre le Cancer Côte d'Or, Nièvre, Saône et Loire, the Association pour la Recherche contre le Cancer (ARC) and the Conseil Régional de Bourgogne.
PY - 2009/9/15
Y1 - 2009/9/15
N2 - 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.
AB - 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.
KW - Evolution
KW - Gene duplication
KW - Inducible nitric oxide synthase
KW - Synteny
KW - Teleostei
UR - http://www.scopus.com/inward/record.url?scp=67650656653&partnerID=8YFLogxK
U2 - 10.1016/j.gene.2009.05.016
DO - 10.1016/j.gene.2009.05.016
M3 - Article
C2 - 19501636
AN - SCOPUS:67650656653
SN - 0378-1119
VL - 445
SP - 58
EP - 65
JO - Gene
JF - Gene
IS - 1-2
ER -