The present project aim to develop in vitro techniques which will refine and replace in vivo animal experiments. Following the establishment of genomic and proteomic tools for investigating model organisms, we now have the opportunity to utilise these te chniques for developing novel cell culture systems applicable to a variety of species, including fish, in nutritional, epidemiological, ecotoxicological, metabolic and functional genomic studies.
Specificly, this project will generate knowledge on how the Atlantic cod, Gadus morhua and Atlantic salmon, Salmo salar, are affected by potential dietary stressors (toxicants, undesirables and supplements) by assesing the in vitro responses of important toxicological tissues via gene expression techniques, fo r the elucidation of gene expression biomarkers. Suitable primary culture systems of hepatocytes and intestinal cells will be developed.
For assessment of developmental stressors, the project will use zebrafish and cod embryonic stem (ES) cells as devel opmental and toxicological models. Once marker genes have been identified for model stressors, cultures can be screened for teratogenic effects of emerging pollutants such as brominated flame retardants and pharmaceutical products of increasing concern in aquatic environments. Knowledge of cell differentiation markers and signal transduction pathways will be of importance for creating differentiation-inducing protocols directing ES cells into specific tissues, thus extending the number of in vitro methods for nutrigenomics and toxicogenomics studies in future.
The development of a GC, LCMS(n) method for analysing polyunsaturated fatty acids and eicosanoids in cells (particularly those of the immune system) in combination with exploration of genomic tech niques will be a powerful tool. This should enable determination of the complex interplay between dietary factors and physiology, and will be applicable to both fish and mammalian research