T2 PM 'Omic' Technologies: Current and Future Applications to Environmental Toxicology (Part 2)
Tuesday, 15 November 2005: 1:50 PM - 5:30 PM in Ballroom 2

(VAN-1117-854241) Rainbow trout cDNA microarray : a tool for assessing contaminants in the aquatic environment.

Osachoff, H¹, Bruno, J¹, Skirrow, R¹, Vijayan, M², Helbing, C³, Mommsen, T³, van Aggelen, G¹, ¹ Environment Canada, North Vancouver, BC, Canada² University of Waterloo, Waterloo, Ont, Canada³ University of Victoria, Victoria, BC, Canada

ABSTRACT- Microarrays have become a common tool for evaluating an organisms changes in gene expression following exposure to contaminants. A rainbow trout (RBT) cDNA microarray has been developed in-house that can be used with additional Pacific salmonid species, such as chinook (O. tshawytscha) and coho (O. kisutch). This microarray couples the field of toxicology with genomics and has been used to examine gene expression changes in rainbow trout exposed to a variety of contaminants, including endocrine disruptors. The RBT microarray comprises 207 cDNA gene transcripts representing broad gene classes such as growth, immune response, metabolism, oncogenesis, transcription and reproduction. The gene transcripts were isolated from rainbow trout tissues using primers designed to obtain unique 450-550 bp pieces and printed on aminosilane-coated Matrix ez-rays^TM (glass slides). The cDNA is directly labeled using Cy dyes^TM (Amersham BioSciences). Studies incorporate the reference experimental design approach, which uses our in-house produced Universal DNA Reference (UDR) material. The UDR is a mixture of each gene printed on the microarray and it is labeled with one of the Cy dyes^TM (typically Cy3^TM) and hybridized to every microarray slide opposite an experimental sample (for example, Cy5^TM labeled cDNA of treated or exposed fish). This approach enables comparison across multiple experiments that include various types of contaminants. Our bioinformatics data analysis system includes image processing with ImaGene® and data acquisition with GeneSpring.

Key words: microarray, rainbow trout