Expression and possible functional roles of cytochromes P450 2J1 (zfCyp 2J1) in zebrafish.

2J (Cyp2J) subfamilies are recognized as catalysts of arachidonic acid metabolism in extrahepatic tissues of many species. However, to date, no P450 2J have been identified in zebrafish. We describe here a zfCyp2J1 cDNA which encodes a putative protein of 496 amino acids and shares 51%, 51%, 50%, 51% and 50% identity with mouse Cyp2J6, rabbit Cyp2J1, human Cyp2J2, cow Cyp2J2, and rat Cyp2J4, respectively. Despite detectable levels of expression by RT-PCR, no expression was shown by in situ hybridization using whole-mount tissues of the embryos. Gene-specific knockdown by antisense morpholino oligonucleotide had no phenotypic effect on embryonic development. However, over-expression of zfCyp2J1 by injection of the embryos with the cDNA resulted in substantial dose-dependent morphological defects. With adult zebrafish, whole-mount in situ hybridization showed that zfCyp2J1 was expressed predominantly in the brain and gonads. A semi-quantitative RT-PCR analysis further revealed that the zfCyp2J1 transcript was also expressed in the ovary, testis, heart, liver, and kidney. High levels of zfCyp2J1 mRNA were evident in primary growth stage (stage I) oocytes and cortical alveolus stage (stage II) oocytes but nearly undetectable in stage III and matured oocytes. These results suggest that zfCyp2J1 may not be involved in zebrafish embryogenesis but may rather play an important role in the functioning of brain and gonads of the adults. In addition, zfCyp2J1 may play a particularly crucial role in early oocyte maturation.

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