Development and validation of a fathead minnow liver cell line for environmental testing

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Primary industries such as forestry, pulp and paper manufacturing and municipal waste treatment facilities employ significant numbers of Canadians. Their operations use large amounts of water and when they return it to the environment it may contain chemicals that affect the ability of animals to successfully reproduce. The fathead minnow is a model species used in experiments to predict the health of other animals in waterways.  Industry and government regulatory agencies use live minnows to screen aquatic effluents for toxicity.  However, these tests are expensive and cumbersome. The primary organ in vertebrates involved in detoxification of environmental contaminants is the liver, and it is very sensitive to low levels of thesecompounds. It is known that toxin-responsive genes are activated or repressed in response to challenge by toxicants, but the identity of many of these genes is unknown. We are currently performing this identification in minnow livers, using a technique to examine gene expression known as DNA microarrays. To simplify and lessen the cost of live testing, we propose to develop a fathead minnow liver cell culture. These cells will be grown in the laboratory and will likely possess the same toxicant-responsive characteristics as cells from livers in fish. However, they will be much easier to grow and use for routine toxicology testing. Some preliminary work has suggested the adoption of cell lines for environmental monitoring, but this project will be among the first to compare gene expression in cell versus intact tissue. This is a necessary prerequisite prior to adopting this technology. Our goal is to validate the use of these cells as a replacement for testing using fish. Thedevelopment of this test will lower the cost of environmental monitoring for Canadian industry, thus helping to safeguard jobs in sectors of critical economic importance to the Canadian economy. Adoption of this test will also reduce the number of lab animals used for toxicology testing, which is a common goal of regulatory agencies worldwide.