PARENT SESSION
PW1 - Molecular/ Cellular Toxicology
Wednesday, 20 November 2002
8:00 AM to 6:30 PM
Exhibit Hall

(P707) Measurement of Copper-Induced Stress in Pimephales promelas Using Gene Expression Profiles.

Lewis, Solange^*,1, Keller, Stephen¹, Lattier, David², Lazorchak, James², Smith, Mark³, ¹ Department of Biological Sciences, University of Cincinnati, Cincinnati, OH, USA² USEPA, 26 West Martin Luther King Drive, Cincinnati, OH, USA³ SoBran, Inc., Cincinnati, OH, USA

ABSTRACT- We are characterizing the gene expression profiles of larval Pimephales promelas in response to varying doses of copper stress. Up to 48-hr old fathead minnow larvae were exposed to copper concentrations of 50 g/L, 125 g/L and 200 g/L over a duration of 48 hours. Survivors from the treated and control groups were collected in duplicate, RNA was isolated, converted to cDNA and then analyzed by the differential display technique using fluorescent anchored primers in combination with random arbitrary primers. Twenty-nine different primer combinations have generated 1379 cDNAs. One hundred and thirty-one cDNAs have been collected as up-regulation candidate genes, of which 42 showed concentration-dependence. One hundred and eighty-one cDNAs were collected as down-regulation candidate genes, 91 of which were dose-dependent. Approximately one percent of the bands were judged to be artifactual because they were not always present in replicate samples. At present, the candidate genes are in the process of being sequenced and some have been identified by homology. Further experiments are in progress utilizing zinc and heat as stressors in order to determine whether the differentially expressed transcripts are copper-specific or are affected by general stress.

Key words: fathead minnow, differential gene expression, copper, stress