Abstract
Studies examined the effects of different culture conditions for the brain, gonads (testis and ovary), and liver of adult male and Female Fathead Minnow (Pimephales promelas) (FHM) during a 28 day assessment of viability and functionality. Five parameters were tested including media pH, incubation temperature, media composition, media exchange rate, and substrate. Within the ranges tested, incubation temperature was the most influential on tissue viability with dramatic improvements using cooler incubation temperatures (<18°C) over the entire 28 day test period. Tissues remained equally viable in five different common culture media, although highest viability with each medium was observed with a weekly media exchange over more frequent exchanges, alginate over polystyrene, and a media pH near physiological values. Use of these combined conditions resulted in continual function of testis and ovarian tissue over the entire 28 days, as indicated by continued production of 11-ketotestosterone (11-KT) by testes and estradiol (E2) by ovaries. Liver function was maintained through seven days, although vitellogenin production in response to added E2 eventually decreased over time. Consistent tissue viability over time periods commonly used for in vivo studies will enhance the link of in vitro tissue toxicology studies to whole-fish and population level impacts.
Highlights
In vitro testing and its link to whole animal biology are becoming increasingly important in predictive ecotoxicology [1]
Fathead minnow (Pimephales promelas) is a common fish species used for ecological monitoring and toxicity testing due to its ease of use as a model species for reproduction and behavior (31,32)
Our goal was to define common culture conditions that satisfy the needs of all hypothalamus (brain)-pituitary-gonadal axis and liver (HPG-L) tissue types simultaneously
Summary
In vitro testing and its link to whole animal biology are becoming increasingly important in predictive ecotoxicology [1]. Better chemical screening systems are necessary to connect adverse outcomes of in vitro studies to population level effects through a mode of action [2]. Chemicals that disrupt the HPG-L endocrine system (endocrine disrupting compounds or EDCs) can have severe impacts on population survival of fish and other animals [5]. Due to the conservation of this system among vertebrates, extrapolating effects of EDCs on fish endocrine physiology can identify potential impact of these contaminants on humans when mechanisms are known. Since thousands of chemicals exist and many more are being introduced, there is an urgent need to determine the reproductive or endocrine disrupting potential of chemicals to which humans and the environment are exposed
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