High-Content and Semi-Automated Quantification of Responses to Estrogenic Chemicals Using a Novel Translucent Transgenic Zebrafish

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• 作者：Jon M. Green ; Jeremy Metz ; Okhyun Lee ; Maciej Trznadel ; Aya Takesono ; A. Ross Brown ; Stewart F. Owen ; Tetsuhiro Kudoh ; Charles R. Tyler
• 刊名：Environmental Science & Technology
• 出版年：2016
• 出版时间：June 21, 2016
• 年：2016
• 卷：50
• 期：12
• 页码：6536-6545
• 全文大小：491K
• 年卷期：0
• ISSN：1520-5851

文摘

Rapid embryogenesis, together with genetic similarities with mammals, and the desire to reduce mammalian testing, are major incentives for using the zebrafish model in chemical screening and testing. Transgenic zebrafish, engineered for identifying target gene expression through expression of fluorophores, have considerable potential for both high-content and high-throughput testing of chemicals for endocrine activity. Here we generated an estrogen responsive transgenic zebrafish model in a pigment-free “Casper” phenotype, facilitating identification of target tissues and quantification of these responses in whole intact fish. Using the ERE-GFP-Casper model we show chemical type and concentration dependence for green fluorescent protein (GFP) induction and both spatial and temporal responses for different environmental estrogens tested. We also developed a semiautomated (ArrayScan) imaging and image analysis system that we applied to quantify whole body fluorescence responses for a range of different estrogenic chemicals in the new transgenic zebrafish model. The zebrafish model developed provides a sensitive and highly integrative system for identifying estrogenic chemicals, their target tissues and effect concentrations for exposures in real time and across different life stages. It thus has application for chemical screening to better direct health effects analysis of environmental estrogens and for investigating the functional roles of estrogens in vertebrates.