Detection of the anti-androgenic effect of endocrine disrupting environmental contaminants using in vivo and in vitro assays in the three-spined stickleback

Jolly, Cecile; Katsiadaki, Ioanna; Morris, Steve; Le Belle, Nadine; Dufour, Sylvie; Mayer, Ian; Pottinger, Tom G.; Scott, Alexander P.. 2009 Detection of the anti-androgenic effect of endocrine disrupting environmental contaminants using in vivo and in vitro assays in the three-spined stickleback. Aquatic Toxicology, 92 (4). 228-239.

Before downloading, please read NORA policies.

Download (865kB)


We have previously developed a novel in vitro assay that utilises cultures of primed female stickleback kidney cells for the screening of potential androgenic and anti-androgenic environmental contaminants. Stickleback kidney cells are natural targets for steroid hormones and are able to produce a protein, spiggin, in response to androgenic stimulation. We undertook a combined in vivo/in vitro study where we used the magnitude of spiggin production as an endpoint to test the anti-androgenic properties of the pharmaceutical androgen antagonist flutamide and three environmental contaminants: the organophosphate insecticide fenitrothion, the urea-based herbicide linuron and the fungicide vinclozolin. In vitro, kidney cells were exposed to a range of concentrations [from 10-14M (2.5pg/L) up to 10-6M (280μg/L)] of the test compounds alone for determining agonist activities, or together with 10-8M (3μg/L) dihydrotestosterone (DHT) for determining antagonist activities. An in vivo flow-trough aquarium-based study was carried out in parallel. Female sticklebacks were exposed to a range of concentrations of the same chemicals alone or in combination with DHT (5μg/L) for 21 days. All of the compounds significantly inhibited DHT-induced spiggin production in a concentration-dependent manner in both the in vitro (FN≥FL≥LN>VZ) and in vivo (FN>FL≥VZ>LN) assays. Fenitrothion and flutamide inhibited spiggin production in vitro at a concentration as low as 10-12M (P<0.05), while linuron and vinclozolin inhibited DHT-induced spiggin production at concentrations of 10-10M (P<0.05) and 10-6M (P<0.001) respectively. Similarly, fenitrothion and flutamide were the most potent chemicals in vivo and significantly reduced DHT-induced spiggin production at a concentration of 10μg/L and 25μg/L respectively (P<0.01). Both linuron and vinclozolin induced a significant decrease in DHT-induced spiggin production at a concentration of 100μg/L when tested in vivo. In addition, kidney cell primary culture was used to test the (anti-)androgenic effects of the major environmental contaminants: oestradiol (E2), nonylphenol (NP) and bisphenol A (BPA) for the first time in teleosts. We observed that these compounds were able to significantly inhibit spiggin production at high doses (E2: 270μg/L; NP: 2.2μg/L; BPA: 2.3μg/L). When tested in the absence of DHT, none of the compounds showed a significant agonistic activity in either in vivo or in vitro assays. Overall, our data further demonstrate that kidney cell primary culture is a reliable and a sensitive screening tool for the detection of (anti-)androgenic compounds. In addition, our study represents the first attempt to develop a combined in vivo/in vitro screening strategy for assessing the effects of (anti-)androgenic endocrine disrupters.

Item Type: Publication - Article
Digital Object Identifier (DOI):
Programmes: CEH Programmes pre-2009 publications > Water > WA02 Quantifying processes that link water quality and quantity, biota and physical environment > WA02.3 Physico-chemical processes and effects on freshwater biot
UKCEH and CEH Sections/Science Areas: Parr
ISSN: 0166-445X
Additional Information. Not used in RCUK Gateway to Research.: The final version of this article is available from
Additional Keywords: stickleback, anti-androgen, spiggin, endocrine disruption, in vitro tests, in vivo tests
NORA Subject Terms: Biology and Microbiology
Ecology and Environment
Related URLs:
Date made live: 03 Jun 2009 13:48 +0 (UTC)

Actions (login required)

View Item View Item

Document Downloads

Downloads for past 30 days

Downloads per month over past year

More statistics for this item...