Use of vitellogenin mRNA as a biomarker for endocrine disruption in feral and cultured fish
Use of vitellogenin mRNA as a biomarker for endocrine disruption in feral and cultured fish
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DOI：
10.1007/s00216-003-2295-1
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发表时间：
2004-02-01
影响因子：
4.3
通讯作者：
Piña, B
中科院分区：
文献类型：
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作者：
García-Reyero, N;Raldúa, D;Piña, B
在雄鱼的血液和肝脏中存在雌性特异性卵黄蛋白前体卵黄蛋白原被广泛用作内分泌干扰的指标。我们研究了几种鱼类肝脏中卵黄蛋白原mRNA的诱导，无论是在鱼缸中还是在野外捕获。我们的程序需要微量的肝脏样品（低至50 mg），如果使用适当的RNA稳定剂，可以应用于现场样品。我们使用逆转录酶PCR和定量实时PCR来检测和精确定量卵黄蛋白原mRNA水平。暴露于17 β-雌二醇的雄性mummichog（Fundulus heteroclitus）的卵黄蛋白原mRNA水平比未处理的雌性高30倍，用壬基酚处理导致该转录物的诱导较弱但一致。我们还研究了卵黄蛋白原mRNA的水平，在人口的普通鲤鱼（鲤鱼）从阿诺亚河，河流已知的雌激素烷基酚的高水平。结果与该采样点记录的鱼类数据一致。最后，我们还检测到卵黄蛋白原mRNA的倒钩（Barbus graellsi），一个物种，没有卵黄蛋白原序列。使用mRNA定量技术分析野生和养殖鱼类的不同物种打开了更精确的检测和进一步控制污染物对当地动物的有害影响暴露于他们的可能性。
The presence of the female-specific yolk protein precursor vitellogenin in blood and liver from male fish is widely used as an indicator of endocrine disruption. We studied the induction of vitellogenin mRNA in liver from several species of fish, both maintained in fish tanks or captured in the wild. Our procedure requires minute amounts of liver samples (down to 50 mg), and can be applied to field samples if the appropriate RNA-stabilisation agent is used. We used reverse-transcriptase PCR and quantitative real-time PCR for detection and precise quantitation of vitellogenin mRNA levels. Male mummichog (Fundulus heteroclitus) exposed to 17beta-estradiol contained levels of vitellogenin mRNA up to 30 times higher than in untreated females and treatment with nonylphenol resulted in a weak but consistent induction of this transcript. We also studied levels of vitellogenin mRNA in a population of common carp (Cyprinus carpio) from the Anoia river, a river known for its high levels of estrogenic alkylphenols. The results were consistent with recorded data for fish from this sampling site. Finally, we also detected vitellogenin mRNA in barbs (Barbus graellsi), a species for which no vitellogenin sequence was available. The use of mRNA quantitation techniques for analysis of feral and cultured fish of different species opens the possibility of more precise detection and further control of the noxious effects of contaminants on the local fauna exposed to them.