W3 AM Toxicogenomics in Environmental Studies
Wednesday, 16 November 2005: 8:00 AM - 11:40 AM in Ballroom 3

(WIN-1117-565967) The Use DNA Microarray Technology to Identify Biomarkers For Exposure of Fathead Minnow to 2,4-Dinitrotoluene.

Wintz, H.¹, Yoo, J.L.², Gibson, A.², Steevens, J.², Perkins, E.², Holman, P.¹, Loguinov, A.¹, Wong, E.¹, Vulpe, C.¹, ¹ UC-Berkeley, Berkeley, CA, USA² USACE Engineer Research and Development Center, Vicksburg, MS, USA

ABSTRACT- The production of explosive 2,4-DNT and its use in military training activities has resulted in its release to the aquatic environment through various surface water pathways. The aquatic toxicology of the DNT homologues is poorly understood and the potential risks involved in exposure to DNT are virtually unknown. The purpose of this study is to characterize the effects of 2,4-dinitrotoluene and the associated mechanisms of toxicity in the freshwater fish species, fathead minnow (Pimephales promelas), through the use of DNA microarray technology. We have constructed a fathead minnow microarray containing 5,000 anonymous cDNAs. A cDNA library was made using RNA extracted from four different populations of fish at different developmental stages between embryo and adult. 5,000 cDNA were picked from a primary library containing a total of 2 million individual clones, PCR-amplified and spotted in duplicate on glass slides. RNA extracted from the liver of fish exposed to 2,4-DNT (11, 22, 44, 88 mole/L) and from unexposed control fish were converted into cDNA, labeled using Cy3 and Cy5 and hybridized to the cDNA microarrays. Statistical analysis of hybridization signals was used to identify 72 cDNAs that are affected by exposure to different concentrations of 2,4-DNT. An analysis of the expression profiles and the identified genes and their potential role in the response to 2,4-DNT will be presented.

Key words: fathead minnow, microarray, dnt, toxicogenomics