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Patrick Larkin,^1,3 Leroy C. Folmar,² Michael J. Hemmer,² Arianna J. Poston,¹ and Nancy D. Denslow¹

¹Department of Biochemistry and Molecular Biology and Center for Biotechnology, University of Florida, Gainesville, Florida, USA; ²National Health and Environmental Effects Research Laboratory, Gulf Ecological Division, U.S. Environmental Protection Agency, Gulf Breeze, Florida, USA; ³EcoArray LLC, Alachua, Florida, USA

Abstract

A variety of anthropogenic compounds are capable of binding to the estrogen receptor (ER) of vertebrate species. Binding of these chemicals to the ER can interfere with homeostasis by altering normal gene expression patterns. The purpose of this study was to characterize the expression of 30 genes using a sheepshead minnow (Cyprinodon variegatus) cDNA macroarray. Many of the genes on the array were previously identified by differential display reverse transcriptase-polymerase chain reaction to be upregulated or downregulated in sheepshead minnows treated through aqueous exposure to known or suspected estrogenic chemicals. The results of this study show that 17ß-estradiol (E₂) , 17-ethinyl estradiol (EE₂) , diethylstilbestrol (DES) , and methoxychlor (MXC) have similar genetic signatures for the 30 genes examined. The genetic signature of fish treated with p-nonylphenol was identical in pattern to that in fish treated with E₂, EE₂, DES, and MXC except for the additional upregulation of a cDNA clone that shares similarity to ubiquitin-conjugating enzyme 9. Endosulfan produced results that resembled the gene expression patterns of untreated control fish with exception of the upregulation of estrogen receptor and the downregulation of a cDNA clone that shares similarity to 3-hydroxy-3-methylglutaryl-coenzyme A reductase. We show that our estrogen-responsive cDNA macroarray can detect dose-dependent changes in gene expression patterns in fish treated with EE₂. Key words: array, biomarkers, endocrine disruption, estrogen, fish, macroarray. Environ Health Perspect 111:839-846 (2003) .