Trichloroethylene effects on gene expression during cardiac development.

Abstract

Halogenated hydrocarbon exposure is associated with changes in gene expression in adult and embryonic tissue. Our study was undertaken to identify differentially expressed mRNA transcripts in embryonic hearts from Sprague-Dawley rats exposed to trichloroethylene (TCE) or potential bio-transformation products dichloroethylene (DCE) and trichloroacetic acid (TCAA). cDNA subtractive hybridization was used to selectively amplify expressed mRNA obtained from control or halogenated hydrocarbon exposed rat embryos. The doses used were 1100 and 110 ppm (8300 and 830 microM) TCE, 110 and 11 ppm (1100 and 110 microM) DCE, and 27.3 and 2.75 mg/ml (100 and 10 mM) TCAA. Control animals were given distilled drinking water throughout the period of experiments. Sequencing of over 100 clones derived from halogenated hydrocarbon exposed groups resulted in identification of numerous differentially regulated gene sequences. Up-regulated transcripts identified include genes associated with stress response (Hsp 70) and homeostasis (several ribosomal proteins). Down-regulated transcripts include extracellular matrix components (GPI-p137 and vimentin) and Ca(2+) responsive proteins (Serca-2 Ca(2+)-ATPase and beta-catenin). Two possible markers for fetal TCE exposure were identified: Serca-2 Ca(2+)-ATPase and GPI-p137, a GPI-linked protein of unknown function. Differential regulation of expression of both markers by TCE was confirmed by dot blot analysis and semi-quantitative RT-PCR with levels of TCE exposure between 100 and 250 ppb (0.76 and 1.9 microM) sufficient to decrease expression. Sequences down-regulated with TCE exposure appear to be those associated with cellular housekeeping, cell adhesion, and developmental processes, while TCE exposure up-regulates expression of numerous stress response and homeostatic genes.