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巴西鱼类中产生的抗奎氏溪蟾卵黄生成素的特性、特异性和敏感性

Characterization, specificity and sensibility of produced anti-Rhamdia quelen vitellogenin in Brazilian fish species.

作者信息

Moura Costa Daniele Dietrich, Bozza Dandie Antunes, Rizzo Luiz Eduardo, Garcia Juan, Costa Michele Dietrich Moura, de Oliveira Ribeiro Ciro Alberto

机构信息

Laboratório de Toxicologia Celular, Departamento de Biologia Celular, Universidade Federal do Paraná, Rua Francisco H. dos Santos, PO Box 19031, Curitiba, PR, Brazil.

Departamento de Patologia Básica, Universidade Federal do Paraná, PO Box 19031, Curitiba, PR, Brazil.

出版信息

Fish Physiol Biochem. 2016 Dec;42(6):1721-1732. doi: 10.1007/s10695-016-0252-0. Epub 2016 Jun 18.

Abstract

Endocrine-disrupting chemicals (EDCs) are widespread used and can interfere on hormone regulation with adverse consequences for both biota and human. Vitellogenin (vtg) is a yolk precursor protein synthesized by the liver in response to estrogen. In order to characterize the vtg of tropical fish Rhamdia quelen and establish a molecular biomarker, adult male individuals were exposed to 17-β-estradiol (E) for vtg induction and anti-R. quelen vtg polyclonal antibodies production. Vitellogenic female fish were used as positive control group. E-induced vtg was characterized as a glycolipophosphoprotein of high molecular mass with peptide mass fingerprint very similar in E-exposed male and vitellogenic female fish. A polyclonal serum containing anti-R. quelen vtg antibodies was produced and showed high specificity and sensibility to detect the vtg of three fish species: R. quelen, Piaractus mesopotamicus and Prochilodus lineatus. Wildlife and laboratory studies reported that EDCs released into the environment may alter the levels of plasma vtg in male fish, making this protein a valuable biomarker of xenoestrogens exposure. Then, we propose the use of anti-R. quelen vtg as a tool for biomonitoring studies and water quality assessment in Brazil and South American countries where the three fish species occur.

摘要

内分泌干扰化学物质(EDCs)被广泛使用,可干扰激素调节,对生物群和人类都会产生不良后果。卵黄蛋白原(vtg)是肝脏响应雌激素合成的一种卵黄前体蛋白。为了表征热带鱼奎氏溪脂鲤的卵黄蛋白原并建立分子生物标志物,成年雄性个体被暴露于17-β-雌二醇(E)以诱导卵黄蛋白原生成并生产抗奎氏溪脂鲤卵黄蛋白原的多克隆抗体。产卵的雌性鱼用作阳性对照组。E诱导产生的卵黄蛋白原被表征为一种高分子量的糖脂磷蛋白,在暴露于E的雄性鱼和产卵的雌性鱼中,其肽质量指纹非常相似。制备了一种含有抗奎氏溪脂鲤卵黄蛋白原抗体的多克隆血清,该血清对检测三种鱼类的卵黄蛋白原具有高特异性和敏感性,这三种鱼类分别是奎氏溪脂鲤(R. quelen)、南美脂鲤(Piaractus mesopotamicus)和条纹锯唇脂鲤(Prochilodus lineatus)。野生动物和实验室研究报告称,释放到环境中的内分泌干扰化学物质可能会改变雄鱼血浆中卵黄蛋白原的水平,使这种蛋白质成为异源雌激素暴露的一种有价值的生物标志物。因此,我们建议将抗奎氏溪脂鲤卵黄蛋白原用作巴西和南美洲有这三种鱼类分布的国家进行生物监测研究和水质评估的工具。

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