Hepatic gene expression profiling using Genechips in zebrafish exposed to 17alpha-ethynylestradiol.
Genomic, proteomic, and metabolomic technologies continue to receive increasing interest from environmental toxicologists. This interest is due to the great potential of these technologies to identify detailed modes of action and to provide assistance in the evaluation of a contaminant's risk to aquatic organisms. Our experimental model is the zebrafish (Danio rerio) exposed to reference endocrine disrupting compounds in order to investigate compound-induced changes in gene transcript profiles. Adult, female zebrafish were exposed to 0, 15, 40, and 100ng/L of 17alpha-ethynylestradiol (EE2) and concentration and time-dependent changes in hepatic gene expression were examined using Affymetrix GeneChip Zebrafish Genome Microarrays. At 24, 48, and 168h, fish were sacrificed and liver mRNA was extracted for gene expression analysis (24 and 168h only). In an effort to link gene expression changes to effects on higher levels of biological organization, body and ovary weights were measured and blood was collected for measurement of plasma steroid hormones (17beta-estradiol (E2), testosterone (T)) and vitellogenin (VTG) using ELISA. EE2 exposure significantly affected gene expression, GSI, E2, T, and VTG. We observed 1622 genes that were significantly affected (p< or =0.001) in a concentration-dependent manner by EE2 exposure at either 24 or 168h. Gene ontology (GO) analysis revealed that EE2 exposure affected genes involved in hormone metabolism, vitamin A metabolism, steroid binding, sterol metabolism, and cell growth. Plasma VTG was significantly increased at 24, 48, and 168h (p< or =0.05) at 40 and 100ng/L and at 15ng/L at 168h. E2 and T were significantly reduced following EE2 exposure at 48 and 168h. GSI was decreased in a concentration-dependent manner at 168h. In this study, we identified genes involved in a variety of biological processes that have the potential to be used as markers of exposure to estrogenic substances. Future work will evaluate the use of these genes in zebrafish exposed to weak estrogens to determine if these genes are indicative of exposure to estrogens with varying potencies.
Hoffmann
J L
JL
Miami Valley Innovation Center, The Procter and Gamble Company, P.O. Box 538707, Cincinnati, OH 45253-8707, USA.
Torontali
S P
SP
Thomason
R G
RG
Lee
D M
DM
Brill
J L
JL
Price
B B
BB
Carr
G J
GJ
Versteeg
D J
DJ
D000818:Animals; D017931:DNA Primers / Q000737:chemistry; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D004781:Environmental Exposure; D004958:Estradiol / Q000097:blood; D004997:Ethinyl Estradiol / Q000032:analysis / Q000633:toxicity*; D005260:Female; D015870:Gene Expression / Q000187:drug effects*; D020869:Gene Expression Profiling / Q000379:methods*; D008099:Liver / Q000187:drug effects / Q000502:physiology; D008297:Male; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013739:Testosterone / Q000097:blood; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics; D014819:Vitellogenins / Q000097:blood; D014874:Water Pollutants, Chemical / Q000633:toxicity*; D015027:Zebrafish / Q000235:genetics*
D017931:DNA Primers; D014819:Vitellogenins; D014874:Water Pollutants, Chemical; D013739:Testosterone; D004997:Ethinyl Estradiol; D004958:Estradiol