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欧洲海鲈(Dicentrachus labrax)11β-羟化酶的互补DNA:热敏期和性腺发生过程中的基因表达

A cDNA for European sea bass (Dicentrachus labrax) 11beta-hydroxylase: gene expression during the thermosensitive period and gonadogenesis.

作者信息

Socorro Silvia, Martins Rute S, Deloffre Laurence, Mylonas Constantinos C, Canario Adelino V M

机构信息

Centro de Investigação em Ciências da Saúde, Faculdade de Ciências da Saúde, Universidade da Beira Interior, Av. Marquês d'Avila e Bolama, 6200 Covilhã, Portugal.

出版信息

Gen Comp Endocrinol. 2007 Jan 1;150(1):164-73. doi: 10.1016/j.ygcen.2006.07.018. Epub 2006 Sep 7.

Abstract

Steroid P450 11beta-hydroxylase, encoded by the CYP11B gene, is a key mitochondrial enzyme for the production of 11-oxygenated androgens, which have been shown to be potent masculinising steroids in several fish species. In this study we have isolated a CYP11B cDNA of 1903 base pairs from the testis of European sea bass (Dicentrarchus labrax) encoding a predicted protein of 552 amino acids. The amino acid identities to other vertebrate 11beta-hydroxylase proteins ranged from 66% to 72% to other fish; 45% to amphibian and 35-39% to mammalian. Southern blot indicated that a single CYP11B gene is present. Northern blot analysis detected two transcripts in testis and head kidney, one of the same size of the isolated clone and the other of 3.9 kb. Reverse transcriptase-polymerase chain reaction showed abundant mRNA expression only in testis and head kidney, being residual in a range of other tissues. Expression of CYP11B and CYP19A (which encodes for ovarian aromatase) was detected from at least 4 days post-hatching and did not appear to be affected by rearing temperature (15 and 20 degrees C) during the first 60 days, a period in which high temperatures promote masculinisation in European sea bass. Throughout, gonadogenesis (60-300 dph), a highly dimorphic pattern of CYP11B expression was consistent with a role of this gene in testicular development.

摘要

细胞色素P450 11β-羟化酶由CYP11B基因编码,是一种关键的线粒体酶,用于生成11-氧化雄激素,在几种鱼类中,这些雄激素已被证明是强效的雄性化类固醇。在本研究中,我们从欧洲海鲈(Dicentrarchus labrax)的睾丸中分离出一个1903个碱基对的CYP11B cDNA,其编码一个预测的552个氨基酸的蛋白质。与其他脊椎动物11β-羟化酶蛋白的氨基酸同一性,与其他鱼类的范围为66%至72%;与两栖动物为45%;与哺乳动物为35%-39%。Southern印迹表明存在单个CYP11B基因。Northern印迹分析在睾丸和头肾中检测到两种转录本,一种与分离克隆的大小相同,另一种为3.9 kb。逆转录聚合酶链反应显示,仅在睾丸和头肾中有丰富的mRNA表达,在一系列其他组织中为残留表达。CYP11B和CYP19A(编码卵巢芳香化酶)的表达在孵化后至少4天就被检测到,并且在前60天似乎不受饲养温度(15和20摄氏度)的影响,在这一时期高温促进欧洲海鲈的雄性化。在整个性腺发生过程中(60-300日龄),CYP11B表达的高度二态性模式与该基因在睾丸发育中的作用一致。

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