A novel cytochrome p450, zebrafish Cyp26D1, is involved in metabolism of all-trans retinoic acid
A novel cytochrome p450, zebrafish Cyp26D1, is involved in metabolism of all-trans retinoic acid
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DOI：
10.1210/me.2005-0362
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发表时间：
2006-07-01
影响因子：
--
通讯作者：
Zhao, Qingshun
中科院分区：
文献类型：
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作者：
Gu, Xingxing;Xu, Fang;Zhao, Qingshun
维甲酸信号传导是脊椎动物胚胎发育所必需的，其作用主要通过维甲酸（RA）与其RA受体和维甲酸-X受体的结合，而RA在胚胎中的临界浓度和定位由视黄酸脱氢酶（用于RA合成）和细胞色素P450 RA（Cyp 26 s）（用于RA降解）的存在和活性决定。在此之前，我们在斑马鱼中发现了一个新的cyp 26基因（cyp 26 d1），该基因在早期发育的后脑中表达。使用反相HPLC分析，我们在这里显示，斑马鱼Cyp 26 D1在293 T细胞中表达可以代谢全反式RA，9-顺式RA，和13-顺式RA，但不能代谢视黄醇或视黄醇。Cyp 26 D1产生的全反式RA代谢产物与Cyp 26 A1相同，主要为4-羟基全反式RA和4-氧代全反式RA。通过对斑马鱼胚胎进行mRNA显微注射，我们发现Cyp 26 D1基因在斑马鱼胚胎中的过表达不仅导致注射胚胎的第一体节与第5菱形节之间的距离比对照胚胎短，而且还导致注射胚胎的体节发生的左右不对称。这些变化是相似的cyp 26 a1在斑马鱼胚胎中的过度表达所造成的，这导致从处理胚胎与1 μ M 4-二乙基氨基苯甲醛（视网膜脱氢酶抑制剂），这意味着cyp 26 d1可以拮抗RA活性在体内。总之，我们的体外和体内结果提供了斑马鱼Cyp 26 D1参与RA代谢的直接证据。
Retinoid signaling is essential for development of vertebrate embryos, and its action is mainly through retinoic acid ( RA) binding to its RA receptors and retinoid-X receptors, while the critical concentration and localization of RA in embryos are determined by the presence and activity of retinal dehydrogenases ( for RA synthesis) and cytochrome P450 RAs (Cyp26s) ( for degradation of RA). Previously, we identified a novel cyp26 gene (cyp26d1) in zebrafish that is expressed in hind-brain during early development. Using reverse-phase HPLC analyses, we show here that zebrafish Cyp26D1 expressed in 293T cells could metabolize all-trans RA, 9-cis RA, and 13-cis RA, but could not metabolize retinol or retinal. The metabolites of all-trans RA produced by Cyp26D1 were the same as that produced by Cyp26A1, which are mainly 4-hydroxy-all-trans-RA and 4-oxo-all-trans-RA. Performing mRNA microinjection into zebrafish embryos, we demonstrated that overexpression of Cyp26D1 in embryos not only caused the distance between rhombomere 5 and the first somite of the injected embryos to be shorter than control embryos but also resulted in left-right asymmetry of somitogenesis in the injected embryos. These alterations were similar to those caused by the overexpression of cyp26a1 in zebrafish embryos and to that which resulted from treating embryos with 1 mu M 4-diethylamino-benzaldehyde ( retinal dehydrogenase inhibitor), implying that cyp26d1 can antagonize RA activity in vivo. Together, our in vitro and in vivo results provided direct evidence that zebrafish Cyp26D1 is involved in RA metabolism.