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大菱鲆(Scophthalmus maximus)雌激素受体的分子特征及定量分析

Molecular characterization and quantification of estrogen receptors in turbot (Scophthalmus maximus).

作者信息

Hu Peng, Meng Zhen, Jia Yudong

机构信息

Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology, Qingdao 266071, China.

Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology, Qingdao 266071, China.

出版信息

Gen Comp Endocrinol. 2018 Feb 1;257:38-49. doi: 10.1016/j.ygcen.2017.01.003. Epub 2017 Jan 10.

DOI:10.1016/j.ygcen.2017.01.003
PMID:28087301
Abstract

Estrogens regulate various reproductive processes via estrogen receptor (ER)-mediated signaling pathway in vertebrates. In this study, full-length sequences coding for ERα, ERβ1 and ERβ2 were isolated from female turbot (Scophthalmus maximus) by homology cloning and a strategy based on rapid amplification of cDNA end-polymerase chain reaction (RACE-PCR). The nucleotide and amino acid sequences of turbot ERs showed high homologies with the corresponding sequences of other fish species and significant homology with the Japanese flounder (Paralichthys olivaceus) and the European sea bass (Dicentrarchus labrax). Turbot ERs contained six typical nuclear receptor-characteristic domains and exhibited high evolutionary conservation in the functional domains. Quantitative real-time polymerase chain reaction analysis revealed that the erα and erβ (β1, β2) mRNAs were abundant in the liver and ovary, respectively. Furthermore, hepatic mRNA levels of erα and vitellogenin (vtg) were found increased gradually from pre-vitellogenesis to late-vitellogenesis stages, with the highest values observed at the late-vitellogenesis stage, and then decreased from migratory-nucleus to atresia stages. However, mRNA levels of erα in the ovary remained unchanged during ovarian development. Hepatosomatic index, gonadosomatic index, serum estradiol-17β and the mRNA levels of erβ1 and erβ2 in the ovary manifested results similar to the expression of erα mRNAs in the liver. These findings indicated that ERα is mainly involved in hepatic vitellogenesis, and ERβs may play crucial roles to regulate ovarian development in turbot. Overall, this study improves understanding of the physiological functions of turbot ERs, which will be valuable for fish reproduction and broodstock management.

摘要

雌激素通过雌激素受体(ER)介导的信号通路调节脊椎动物的各种生殖过程。在本研究中,通过同源克隆以及基于cDNA末端快速扩增-聚合酶链反应(RACE-PCR)的策略,从雌性大菱鲆(Scophthalmus maximus)中分离出编码ERα、ERβ1和ERβ2的全长序列。大菱鲆ERs的核苷酸和氨基酸序列与其他鱼类的相应序列具有高度同源性,与牙鲆(Paralichthys olivaceus)和欧洲海鲈(Dicentrarchus labrax)的相应序列具有显著同源性。大菱鲆ERs包含六个典型的核受体特征结构域,并且在功能结构域中表现出高度的进化保守性。实时定量聚合酶链反应分析表明,erα和erβ(β1、β2)mRNA分别在肝脏和卵巢中大量存在。此外,发现从卵黄发生前期到卵黄发生后期,肝脏中erα和卵黄蛋白原(vtg)的mRNA水平逐渐升高,在卵黄发生后期达到最高值,然后从迁移核期到闭锁期逐渐下降。然而,在卵巢发育过程中,卵巢中erα的mRNA水平保持不变。肝体指数、性腺指数、血清雌二醇-17β以及卵巢中erβ1和erβ2的mRNA水平表现出与肝脏中erα mRNA表达相似的结果。这些发现表明,ERα主要参与肝脏卵黄生成,而ERβs可能在调节大菱鲆卵巢发育中起关键作用。总体而言,本研究增进了对大菱鲆ERs生理功能的理解,这对于鱼类繁殖和亲鱼管理具有重要价值。

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