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GENE EXPRESSION PATTERNS OF CD-1 DAY-8 EMBRYO CULTURES EXPOSED TO BROMOCHLORO ACETIC ACID
Citation:
Karoly, E. D., E S. Hunter III, AND J E. Schmid. GENE EXPRESSION PATTERNS OF CD-1 DAY-8 EMBRYO CULTURES EXPOSED TO BROMOCHLORO ACETIC ACID. Presented at Gordon Research Conference, Lewiston, ME, June 22-27, 2003.
Description:
Gene expression patterns of CD-1 day-8 embryo cultures exposed to bromochloro acetic acid
Edward D. Karoly?*, Judith E. Schmid* and E. Sidney Hunter III*
?Curriculum in Toxicology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina and *Reproductive Toxicology Division, National Health and Environmental Effects Research Laboratory, Office of Research and Development, U.S. EPA, Research Triangle Park, North Carolina 27711
An unintended consequence of chemical disinfection of municipal drinking water has been the production of disinfection by-products (DBPs) generated through the interaction of the chemical disinfectants and naturally occurring organic material present in the water. Bromochloroacetic acid (BCA), a common and abundantly found DBP included within the family of haloacetic acids, is a reproductive toxicant in animal models. In order to examine altered gene expression in mouse embryos after exposure to BCA, CD-1 mouse 8-day embryos were cultured in 300?M BCA for 0, 1, 3 and 6h. Additional embryos were exposed to 30 and 3?M BCA for 6h. Following exposure, embryos were terminated and heads were pooled for total RNA isolation. Total RNA was reverse transcribed with amino allyl-dUTPand indirectly labeled with Cy3 and Cy5. Fluorescently labeled cDNA from treated and untreated samples were hybridized against a standard reference sample onto custom glass slide arrays spotted with 70mer oligonucleotide probesfrom the Operon Array-Ready Oligo Set for the mouse genome version 2 (Qiagen Operon, Valencia, CA). Slides were scanned and quantitated with ScanArray Express (PerkinElmer Life Sciences, Boston, MA). The data were normalized by slide using an intensity dependent local regression (SAS Proc Loess). Analysis of all slides identified 3,473 genes as deviating from the control mean value (? 2 STD). Analysis of the 0 and 300?M BCA time course identified 2,721 significant genes that were separated into 13 clusters using K-means clustering(SAS Proc Fastclus). Further bioinformatic analysis of this gene expression data set is currently in progress. This is an abstract of a proposed presentation and does not necessarily reflect EPA policy. Funded by EPA/UNC Toxicology research program, training agreement CT 827206 with the Curriculum of Toxicology, University of North Carolina at Chapel Hill, North Carolina.