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中华鳖(Pelodiscus sinensis)cyp19a 基因的表达与鉴定及其对雌二醇/来曲唑暴露的响应

Expression and characterization of the cyp19a gene and its responses to estradiol/letrozole exposure in Chinese soft-shelled turtle (Pelodiscus sinensis).

机构信息

Key Laboratory of Aquatic Genomics, Ministry of Agriculture and Rural Affairs, Yangtze River Fisheries Research Institute, Chinese Academy of Fisheries Science, Hubei, China.

Devision of Fish Genetics and Breeding, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Hubei, China.

出版信息

Mol Reprod Dev. 2019 May;86(5):480-490. doi: 10.1002/mrd.23126. Epub 2019 Feb 18.

Abstract

Cytochrome P450 aromatase (CYP19) catalyzes the conversion of androgens to estrogens and is critical in sex differentiation. CYP19 exists as the ovarian type and brain type. Herein, we cloned the full-length ovarian cyp19a gene from the Chinese soft-shelled turtle, Pelodiscus sinensis (pscyp19a). We determined the distribution of pscyp19a in adult tissue and evaluated its expression during embryonic development, following treatment with 17β-estradiol (E2) or letrozole (LE). The pscyp19a complementary DNA is 2,285 bp in length and comprises a 1,512 bp open reading frame that encodes a protein of 503 AA. The nucleotide sequence and amino acid of pscyp19a shared significant identity with other vertebrate sequences. Expression of pscyp19a was high in the ovary (p < 0.01), and exhibited modest expression in the female brain and intestine. Expression of pscyp19a displayed significant differences between sexes during early embryo development stages; expression increased gradually during embryonic development in females, but the opposite trend was observed in males. Female embryos treated with different concentrations of E2 and LE displayed altered pscyp19a expression compared with untreated individuals, and E2 clearly induced pscyp19a expression. These results indicate that pscyp19a gene plays important roles in early developmental stages in Chinese soft-shelled turtle, and may assist future studies on sex differentiation and sex control in this and similar species.

摘要

细胞色素 P450 芳香酶(CYP19)催化雄激素向雌激素的转化,在性别分化中至关重要。CYP19 存在卵巢型和脑型。本文从中华鳖(Pelodiscus sinensis)中克隆了全长卵巢 cyp19a 基因(pscyp19a)。我们测定了 pscyp19a 在成年组织中的分布,并评估了其在胚胎发育过程中的表达,以及 17β-雌二醇(E2)或来曲唑(LE)处理后的表达情况。pscyp19a cDNA 长 2285bp,包含一个 1512bp 的开放阅读框,编码一个 503 个氨基酸的蛋白质。pscyp19a 的核苷酸序列和氨基酸与其他脊椎动物序列具有显著的同源性。pscyp19a 在卵巢中表达量较高(p<0.01),在雌性大脑和肠道中也有中等表达。在胚胎发育早期,pscyp19a 在两性之间的表达存在显著差异;在雌性中,随着胚胎发育逐渐增加,但在雄性中则呈现相反的趋势。与未处理个体相比,用不同浓度的 E2 和 LE 处理的雌性胚胎的 pscyp19a 表达发生改变,E2 明显诱导了 pscyp19a 的表达。这些结果表明,pscyp19a 基因在中华鳖的早期发育阶段发挥重要作用,并可能有助于未来对该种及类似物种的性别分化和性别控制的研究。

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