|
|
|
Adapting Microarray Technology for Use in Ecotoxicogenomics
 |
|
| Abstract: | Ecotoxicogenomics includes research to identify differential gene expression in laboratory and field animals exposed to toxicants, and ultimately, to link the earliest indicators of exposure to adverse effects in organisms and populations. The USEPA National Exposure Research Laboratory has started to develop a multiple aquatic stressor diagnostic exposure model using fathead minnow (Pimephales promelas) gene sequences in a microarray platform. Because there is very little gene sequence information for the target organism, an approach involving several different methodologies has been developed to examine specific gene expression and to identify stressor-specific patterns of expression over a portion of the expressed genome. Single exposure-specific subtractive cDNA libraries were constructed from adult liver, fry, and embryos to identify differentially expressed genes in minnows exposed to low levels of endocrine disruptor compounds. To complement genes isolated in subtractive arrays, other P. promelas gene sequences were identified by differential display PCR to isolate EDC sensitive genes. In addition, PCR primers based on close relatives of the fathead minnow, the common carp and goldfish, were used to identify homologs of known carp or goldfish genes. By combining these methods for identifying fathead minnow specific sequences, different functional classes of protein-encoding genes are used to build an oligo microarray that encompasses a sampling of the expressed genome. Single and multiple exposures are assessed at the oligo microarray level to gain a better understanding of relative bioavailability of environmental stressors present in mixtures. Thus, the use of microarray technology to examine gene expression promises to be a powerful tool in understanding environmental exposures and their impacts. |
|
| Citation: | Miracle, A. L., D. L. Lattier, R. Flick, T. V. Reddy, and G. P. Toth. Adapting Microarray Technology for Use in Ecotoxicogenomics. Presented at Third International Conference on Pharmaceuticals and Endocrine Disrupting Chemicals, Minneapolis, MN, March 19-21, 2003. |
|
|
| Contact: |
Linda Ransick - (513) 569-7395 or ransick.linda@epa.gov
|
|
| Division: |
Ecological Exposure Research Division |
|
| Branch: |
Molecular Ecology Research Branch |
|
| Product Type: |
Sympos/Conf |
|
| Presented: |
03/19/2003 |
|
|
|
