Journal
ENVIRONMENTAL SCIENCE & TECHNOLOGY
Volume 48, Issue 18, Pages 10919-10928Publisher
AMER CHEMICAL SOC
DOI: 10.1021/es5030977
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Funding
- WatchFrog
- Marie Curie Actions project SME Receptor [PIAP-GA-2008-217877]
- Marie Curie Actions project EDA-EMERGE [FP7-PEOPLE-2011-ITN-290100]
- Ministry of the Environment, Japan through the UK-J collaboration project
- Ministry of Education, Culture, Sports, Science and Technology, Japan [23570085, 24370029]
- Grants-in-Aid for Scientific Research [23570085] Funding Source: KAKEN
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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? 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 ?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.
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