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欧洲鳗鲡(Anguilla anguilla)细胞色素P450芳香化酶基因(CYP19)的克隆与发育表达

Cloning and developmental expression of the cytochrome P450 aromatase gene (CYP19) in the European eel (Anguilla anguilla).

作者信息

Tzchori Itai, Degani Gad, Hurvitz Avshalom, Moav Boaz

机构信息

Department of Zoology, Tel Aviv University, Tel Aviv 69978, Israel.

出版信息

Gen Comp Endocrinol. 2004 Sep 15;138(3):271-80. doi: 10.1016/j.ygcen.2004.06.007.

DOI:10.1016/j.ygcen.2004.06.007
PMID:15364210
Abstract

To characterize the involvement of the aromatase gene during the process of sex determination in the European eel (Anguilla anguilla), the expression of its gonadal form was determined during various developmental stages. The cloned cDNA from the European eel gonad (EeCYP19) contains an open reading frame of 1539 bp, encoding a deduced protein of 513 residues. The predicted amino acid sequence shows 97% identity with that of the Japanese eel, and 59-69% of identity with those of the CYP19 gonadal and brain forms of other teleost fish. Two potential initiation sites (ATG) were found downstream of the first ATG codon. A fluorescent-based method of real-time PCR was developed to quantify EeCYP19 expression. The expression levels of EeCYP19 in the gonads of adult males were approximately 12- and 30-fold lower than the levels in adult females and juvenile eels previously treated with E2, respectively. Expression of aromatase was found only in a single specimen in the control group. In contrast, no difference was found among sexes in the aromatase expression in the brain. Treatment with aromatase inhibitor (AI) of juvenile eel resulted in the total loss of aromatase expression in the gonads and brains. The results of this work revealed that AI treatment not only reduces the synthesis of estradiol, but reduces the expression levels of EeCYP19 as well. No evidence for the presence of a distinct extra-gonadal (brain) form of aromatase in the European eel could be provided.

摘要

为了表征芳香化酶基因在欧洲鳗鲡(Anguilla anguilla)性别决定过程中的作用,我们测定了其性腺形式在不同发育阶段的表达情况。从欧洲鳗鲡性腺克隆的cDNA(EeCYP19)含有一个1539 bp的开放阅读框,编码一个由513个氨基酸残基组成的推导蛋白。预测的氨基酸序列与日本鳗鲡的序列有97%的同一性,与其他硬骨鱼类CYP19性腺和脑形式的序列有59 - 69%的同一性。在第一个ATG密码子下游发现了两个潜在的起始位点(ATG)。开发了一种基于荧光的实时PCR方法来定量EeCYP19的表达。成年雄性性腺中EeCYP19的表达水平分别比成年雌性和先前用E2处理过的幼鳗低约12倍和30倍。在对照组中,仅在一个样本中发现了芳香化酶的表达。相比之下,脑中芳香化酶的表达在性别之间没有差异。用芳香化酶抑制剂(AI)处理幼鳗导致性腺和脑中芳香化酶表达完全丧失。这项工作的结果表明,AI处理不仅降低了雌二醇的合成,还降低了EeCYP19的表达水平。没有证据表明欧洲鳗鲡存在一种独特的性腺外(脑)形式的芳香化酶。

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