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TRANSCRIPTIONAL RESPONSES OF MOUSE EMBRYO CULTURES EXPOSED TO BROMOCHLOROACETIC ACID
Citation:
Karoly, E. D., J E. Schmid, AND E I. Hunter. TRANSCRIPTIONAL RESPONSES OF MOUSE EMBRYO CULTURES EXPOSED TO BROMOCHLOROACETIC ACID. Presented at Society of Toxicology, Baltimore, MD, March 21-25, 2004.
Description:
Transcriptional responses of mouse embryo cultures exposed to bromochloroacetic 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, RTP, North Carolina 27711
A major class of disinfection by-products (DBPs) found in municipal drinking water are the haloacetic acids. Bromochloroacetic acid (BCA), a common and abundantly found DBP included within the family of haloacetic acids, is a reproductive toxicant in animal models. The mechanism of haloacetic acid embryotoxicity has not been defined, but may be mediated in part through the inhibition of protein kinases. In order to examine altered gene expression in mouse embryos following exposure to BCA, CD-1 mouse embryos (3 to 6 somite stage) were cultured in 300?M BCA for 0, 1, 3 and 6h. Additionally, embryos were exposed to 30 and 3?M BCA for 6h. Following exposure, embryos were terminated and a total of 30 heads per treatment were pooled for RNA isolation. Total RNA was reverse transcribed with amino allyl-dUTP and indirectly labeled with Cy3 and Cy5. Fluorescently labeled cDNA from treated and untreated samples were labeled with Cy5 and hybridized against a Cy3 labeled standard reference sample onto custom glass slide arrays. Arrays were spotted with 17,388 70mer oligonucleotide probes from the Operon Array-Ready Oligo Set for the mouse genome version 2 (Qiagen Operon, Valencia, CA). The data were normalized by slide using an intensity dependent local regression (SAS Proc Loess). Two-way ANOVA identified 276 genes as being differentially expressed across dose and/or time. Cluster analysis and linear models were used to identify the transcriptional responses in mouse embryo culture exposed to BCA and their association with mouse embryotoxicity. 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.