Rapid fluorescent detection of (anti)androgens with spiggin-gfp medaka

Anthony Sébillot, Pauliina Damdimopoulou, Yukiko Ogino, Petra Spirhanzlova, Shinichi Miyagawa, David Du Pasquier, Nora Mouatassim, Taisen Iguchi, Gregory F. Lemkine, Barbara A. Demeneix, Andrew J. Tindall

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21 Citations (Scopus)

Abstract

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 threespined 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 μg/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.

Original languageEnglish
Pages (from-to)10919-10928
Number of pages10
JournalEnvironmental Science and Technology
Volume48
Issue number18
DOIs
Publication statusPublished - Sep 16 2014
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Chemistry(all)
  • Environmental Chemistry

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    Sébillot, A., Damdimopoulou, P., Ogino, Y., Spirhanzlova, P., Miyagawa, S., Du Pasquier, D., Mouatassim, N., Iguchi, T., Lemkine, G. F., Demeneix, B. A., & Tindall, A. J. (2014). Rapid fluorescent detection of (anti)androgens with spiggin-gfp medaka. Environmental Science and Technology, 48(18), 10919-10928. https://doi.org/10.1021/es5030977