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生成和鉴定 gsuα:EGFP 转基因斑马鱼用于评估内分泌干扰效应。

Generation and characterization of gsuα:EGFP transgenic zebrafish for evaluating endocrine-disrupting effects.

机构信息

Key Laboratory of Aquatic Biodiversity and Conservation of the Chinese Academy of Sciences, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, Hubei, China; University of Chinese Academy of Sciences, Beijing, China.

Key Laboratory of Aquatic Biodiversity and Conservation of the Chinese Academy of Sciences, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, Hubei, China.

出版信息

Toxicol Appl Pharmacol. 2014 Jul 1;278(1):78-84. doi: 10.1016/j.taap.2014.04.009. Epub 2014 Apr 18.

Abstract

The glycoprotein subunit α (gsuα) gene encodes the shared α subunit of the three pituitary heterodimeric glycoprotein hormones: follicle-stimulating hormone β (Fshβ), luteinizing hormone β (Lhβ) and thyroid stimulating hormone β (Tshβ). In our current study, we identified and characterized the promoter region of zebrafish gsuα and generated a stable gsuα:EGFP transgenic line, which recapitulated the endogenous gsuα expression in the early developing pituitary gland. A relatively conserved regulatory element set is presented in the promoter regions of zebrafish and three other known mammalian gsuα promoters. Our results also demonstrated that the expression patterns of the gsuα:EGFP transgene were all identical to those expression patterns of the endogenous gsuα expression in the pituitary tissue when our transgenic fish were treated with various endocrine chemicals, including forskolin (FSK), SP600125, trichostatin A (TSA), KClO4, dexamethasone (Dex), β-estradiol and progesterone. Thus, this gsuα:EGFP transgenic fish reporter line provides another valuable tool for investigating the lineage development of gsuα-expressing gonadotrophins and the coordinated regulation of various glycoprotein hormone subunit genes. These reporter fish can serve as a novel platform to perform screenings of endocrine-disrupting chemicals (EDCs) in vivo as well.

摘要

糖蛋白亚基 α (gsuα) 基因编码三种垂体异二聚体糖蛋白激素的共享 α 亚基:卵泡刺激素 β (Fshβ)、黄体生成素 β (Lhβ) 和促甲状腺素 β (Tshβ)。在我们目前的研究中,我们鉴定并描绘了斑马鱼 gsuα 的启动子区域,并生成了一个稳定的 gsuα:EGFP 转基因系,该系在早期发育的垂体中 recapitulated 内源性 gsuα 的表达。在斑马鱼和其他三种已知的哺乳动物 gsuα 启动子的启动子区域中呈现了一组相对保守的调控元件。我们的结果还表明,当我们的转基因鱼用各种内分泌化学物质处理时,包括 forskolin (FSK)、SP600125、trichostatin A (TSA)、KClO4、地塞米松 (Dex)、β-雌二醇和孕酮,gsuα:EGFP 转基因的表达模式与内源性 gsuα 在垂体组织中的表达模式完全相同。因此,这种 gsuα:EGFP 转基因鱼报告系为研究 gsuα 表达的促性腺激素的谱系发育以及各种糖蛋白激素亚基基因的协调调控提供了另一种有价值的工具。这些报告鱼还可以作为一种新的平台,在体内进行内分泌干扰化学物质 (EDCs) 的筛选。

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