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PW05 Genomics and Proteomics
(PW084) Differential display of hepatic mRNA from Fundulus heteroclitus inhabiting a Superfund estuary.
Meyer, J1, Freedman, J1, Di Giulio, R1, 1 Duke University, Durham, NC, 27708-0328
ABSTRACT- Fundulus heteroclitus Atlantic killifish, mummichog) from a highly contaminated site on the Elizabeth River (VA, USA) are resistant to the toxicity of sediment from the site, which is teratogenic to embryos and lethal to larval killifish from relatively pristine reference sites (Ownby et al., 2002; Meyer and Di Giulio, 2003). However, the mechanistic changes that confer resistance to the toxicity of the Elizabeth River sediments are not yet well understood. We are utilizing differential display to identify genes that are differentially expressed in hepatic tissue of killifish adapted to survival in the contaminated Elizabeth River environment, as compared to killifish from a reference site, King's Creek (VA, USA). We compared mRNA expression in 3 pools (2 individuals/pool) of livers per sex per population (12 total samples, corresponding to 24 individuals); livers were taken from wild-caught adults. To date, 24 primer combinations have been used, corresponding to approximately 20% coverage of expressed mRNAs. High interindividual variability and multiple sex-specific differences were observed in mRNA expression, validating the experimental design. 22 population-specific differences have also been identified, and 12 of these cDNAs have been cloned and sequenced. mRNAs identified as upregulated in Elizabeth River killifish include type 2 deiodinase, a serum tributlytin binding protein, and an unidentified mRNA also upregulated in medaka cells after in vitro exposure to UV irradiation. We are currently using additional primer sets and pursuing the cloning and sequencing of additional cDNAs. Research supported by the Office of Naval Research (N00014-00-1-0315, RTD) and the NIEHS-supported Duke University Integrated Toxicology Program (T32-ES-07031-21A1) and Duke University Superfund Basic Research Center (P42 ES10356, RTD).
Key words: adaptation, creosote, differential display, Fundulus heteroclitus
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