Rapid Fluorescent Detection of (Anti)androgens with spiggin-gfp Medaka

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• 作者：Anthony S茅billot ; Pauliina Damdimopoulou ; Yukiko Ogino ; Petra Spirhanzlova ; Shinichi Miyagawa ; David Du Pasquier ; Nora Mouatassim ; Taisen Iguchi ; Gregory F. Lemkine ; Barbara A. Demeneix ; Andrew J. Tindall
• 刊名：Environmental Science & Technology
• 出版年：2014
• 出版时间：September 16, 2014
• 年：2014
• 卷：48
• 期：18
• 页码：10919-10928
• 全文大小：501K
• ISSN：1520-5851

文摘

Widespread environmental antiandrogen contamination has been associated with negative impacts on biodiversity and human health. In particular, many pesticides are antiandrogenic, creating a need for robust and sensitive environmental monitoring. Our aim was to develop a sensitive and specific transgenic medaka (Oryzias latipes) model bearing an androgen responsive fluorescent reporter construct for whole organism-based environmental screening of pro- and antiandrogens. We analyzed the 5鈥?regions of the androgen responsive three-spined stickleback (Gasterosteus aculeatus) spiggin genes in silico, revealing conserved blocks of sequence harboring androgen response elements. Identified putative promoters were cloned upstream of GFP. Germinal transgenesis with spg1-gfp led to stable medaka lines. GFP induction was exclusive to the kidney, the site of spiggin protein production in sticklebacks. Significant GFP expression was induced by three or four-day androgen treatment of newly hatched fry, but not by estrogens, mineralocorticoids, glucocorticoids or progestogens. The model responded dose-dependently to androgens, with highest sensitivity to 17MT (1.5 渭g/L). In addition to flutamide, the biocides fenitrothion, vinclozolin and linuron significantly inhibited 17MT-induced GFP induction, validating the model for detection of antiandrogens. The spg1-gfp medaka model provides a sensitive, specific, and physiologically pertinent biosensor system for analyzing environmental androgen activity.