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三种雌激素受体转录本在莫桑比克罗非鱼(Peters)中的分离和鉴定

Isolation and characterization of three estrogen receptor transcripts in Oreochromis mossambicus (Peters).

机构信息

Ecophysiology Laboratory, Department of Botany and Zoology, University of Stellenbosch, Stellenbosch, South Africa.

出版信息

J Steroid Biochem Mol Biol. 2010 Mar;119(1-2):26-34. doi: 10.1016/j.jsbmb.2009.12.002. Epub 2009 Dec 16.

Abstract

Exposure of aquatic organisms to 17beta-estradiol (E(2)) induces a variety of estrogen-responsive genes, including vitellogenin (vtg)-the precursor protein of egg yolk in oviparous animals and to date the single most used gene product in screening for estrogenic endocrine disruption. Transcription regulation of vtg by E(2) is dependent on binding of the ligand (E(2)) to a specific nuclear receptor (estrogen receptor, ESR) which in turn binds to an estrogen responsive element (ERE) in the promoter of vtg. Since a local tilapiine, Oreochromis mossambicus (Peters), is targeted as a model for estrogenic endocrine disruption in Southern Africa, a platform of knowledge is necessary for the ontogenic and tissue specific behavior of ESR in this species before vtg levels can be interpreted in relation to such endocrine disruption. Therefore, three ESR cDNA sequences (ESR1, ESR2a and ESR2b) in O. mossambicus were isolated and QPCR protocols were developed to ascertain their quantitative transcript levels in adult brain, gonadal and hepatic tissues. ESR1 transcript levels were highest in female liver tissue compared to males and other tissues, whereas the levels for ESR2a and b were not statistically significantly different between male and female tissues. Quantitative gene levels during development demonstrated a sharp increase in ESR1 during the stage of gonad differentiation (50-60 days post-fertilization) in this species. Finally, an induction experiment in adult male liver tissue confirms the upregulation of ESR1 by E(2).

摘要

水生生物暴露于 17β-雌二醇(E(2))会诱导多种雌激素反应基因,包括卵黄蛋白原(vtg)-卵生动物卵黄的前体蛋白,迄今为止是筛选雌激素内分泌干扰物的单一最常用基因产物。E(2)对 vtg 的转录调控依赖于配体(E(2))与特定核受体(雌激素受体,ESR)的结合,该受体反过来与 vtg 启动子中的雌激素反应元件(ERE)结合。由于南部非洲将本地罗非鱼奥利亚罗非鱼(Peters)作为雌激素内分泌干扰的模型,因此在可以根据这种内分泌干扰来解释 vtg 水平之前,需要在这种物种中建立关于 ESR 的个体发生和组织特异性行为的知识库。因此,从奥利亚罗非鱼中分离出三种 ESR cDNA 序列(ESR1、ESR2a 和 ESR2b),并开发了 QPCR 方案来确定它们在成年脑、性腺和肝组织中的定量转录水平。与其他组织相比,ESR1 的转录水平在雌性肝组织中最高,而 ESR2a 和 b 的水平在雄性和雌性组织之间没有统计学上的显著差异。发育过程中的定量基因水平表明,在该物种的性腺分化阶段(受精后 50-60 天),ESR1 急剧增加。最后,在成年雄性肝组织中的诱导实验证实了 E(2)对 ESR1 的上调。

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