Final Report: Species-Specific Endocrine Disruption: PCB- and PAH-Induced Estrogenic Effects

EPA Grant Number: R826301
Title: Species-Specific Endocrine Disruption: PCB- and PAH-Induced Estrogenic Effects
Investigators: Zacharewski, Timothy
Institution: Michigan State University
EPA Project Officer: Turner, Vivian
Project Period: January 1, 1998 through December 31, 2000
Project Amount: $282,998
RFA: Endocrine Disruptors (1997) RFA Text |  Recipients Lists
Research Category: Economics and Decision Sciences , Endocrine Disruptors , Health , Safer Chemicals


This project examined the alleged estrogen receptor-mediated activities of a structurally diverse set of compounds using a battery of species-based in vitro and in vivo assays. The studies were performed using estrogen receptors from a number of species, including human, rodent, bird, frog, reptile and fish, to test the hypothesis that the use of a single surrogate species was inappropriate to identify and assess the estrogenic endocrine disrupting activities of structurally diverse chemicals to human and wildlife health due to divergent amino acid sequences within the estrogen receptor ligand binding domains observed among species.

Summary/Accomplishments (Outputs/Outcomes):

Species-specific responses to structurally diverse synthetic and natural estrogenic compounds (e.g., steroids, pharmaceuticals, phytoestrogens, mycotoxins, organochlorine compounds, PCBs, polyaromatic hydrocarbons, and other industrial chemicals) were investigated using a battery of in vitro and in vivo assays. Competitive ligand binding and report gene assays identified several compounds that exhibited differential interactions with recombinant estrogen receptors (ER) consisting of the D, E, and F domains of the human and , mouse and , chicken, green anole, xenopus and rainbow trout ERs. Qualitatively, all receptor fusion proteins bound the same set of compounds. However, there were significant differences in relative binding affinities between species. In general, reporter gene expression data correlated with competitive binding results. Intriguingly, the rainbow trout ER ligand binding and reporter gene transactivation were found to be temperature dependent and functioned more effectively closer to the physiological temperature. Comparison of the amino acids that lined the binding pocket of the rainbow trout and human ERs identified two conservative substitutions, which were found through reciprocal mutagenesis studies to play a significant role in this temperature dependence. In vivo, the ability of -zearalenol, genistein, and 4-t-octylphenol, three compounds that exhibited differential activity in vitro, to induce vitellogenin mRNA levels in juvenile trout, xenpous and Japanese quail was examined. There was a poor correlation between the in vitro and in vivo activities and striking differences in efficacy and potency between the species examined. Overall, these results do not preclude the use of a single surrogate species for screening purposes. However, the ability to predict in vivo activity based on in vitro data is questionable.

Journal Articles on this Report : 9 Displayed | Download in RIS Format

Other project views: All 50 publications 11 publications in selected types All 10 journal articles
Type Citation Project Document Sources
Journal Article Celius T, Matthews JB, Giesy JP, Zacharewski TR. Quantification of rainbow trout (Oncorhynchus mykiss) zona radiata and vitellogenin mRNA levels using real-time PCR after in vivo treatment with estradiol-17 beta or alpha-zearalenol. Journal of Steroid Biochemistry and Molecular Biology 2000;75(2-3):109-119. R826301 (2000)
R826301 (Final)
not available
Journal Article Fertuck KC, Matthews JB, Zacharewski TR. Hydroxylated benzo[a]pyrene metabolites are responsible for in vitro estrogen receptor-mediated gene expression induced by benzo[a]pyrene, but do not elicit uterotrophic effects in vivo. Toxicological Sciences 2001;59(2):231-240. R826301 (2001)
R826301 (Final)
not available
Journal Article Fertuck KC, Kumar S, Sikka HC, Matthews JB, Zacharewski TR. Interaction of PAH-related compounds with the and isoforms of estrogen receptor. Toxicology Letters 2001;121(3):167-177. R826301 (2001)
R826301 (Final)
R826192 (Final)
not available
Journal Article Huang YW, Matthews JB, Fertuck KC, Zacharewski TR. Use of Xenopus laevis as a model for investigating in vitro and in vivo endocrine disruption in amphibians. Environmental Toxicology and Chemistry 2005;24(8):2002-2009 R826301 (Final)
not available
Journal Article Matthews JB, Twomey K, Zacharewski TR. In vitro and in vivo interactions of bisphenol A and its metabolite, bisphenol A glucuronide, with estrogen receptors alpha and beta. Chemical Research in Toxicology 2001;14(2):149-157 R826301 (Final)
not available
Journal Article Matthews JB, Clemons JH, Zacharewski TR. Reciprocal mutagenesis between human α(L349, M528) and rainbow trout (M317, I496) estrogen receptor residues demonstrates their importance in ligand binding and gene expression at different temperatures. Molecular and Cellular Endocrinology 2001;183(1-2):127-139 R826301 (2001)
R826301 (Final)
not available
Journal Article Matthews JB, Fertuck KC, Celius T, Huang YW, Fong CJ, Zacharewski TR. Ability of structurally diverse natural products and synthetic chemicals to induce gene expression mediated by estrogen receptors from various species. Journal of Steroid Biochemistry and Molecular Biology 2002;82(2-3):181-194 R826301 (Final)
not available
Journal Article Matthews J, Zacharewski T. Differential binding affinities of PCBs, HO-PCBs, and aroclors with recombinant human, rainbow trout (Onchorhynkiss mykiss), and green anole (Anolis carolinensis) estrogen receptors, using a semi-high throughput competitive binding assay. Toxicological Sciences 2000;53(2):326-339. R826301 (1999)
R826301 (Final)
not available
Journal Article Rosen G, O'Bryant E, Matthews J, Zacharewski T, Wade J. Distribution of androgen receptor mRNA expression and immunoreactivity in the brain of the green anole lizard. Journal of Neuroendocrinology 2002;14(1):19-28. R826301 (2001)
R826301 (Final)
not available

Supplemental Keywords:

diethylstilbestrol, genistein, bisphenol A., RFA, Health, Scientific Discipline, Toxics, Waste, Environmental Chemistry, Health Risk Assessment, HAPS, Endocrine Disruptors - Environmental Exposure & Risk, chemical mixtures, endocrine disruptors, Risk Assessments, Biochemistry, Children's Health, Endocrine Disruptors - Human Health, adverse outcomes, complex mixtures, endocrine disrupting chemicals, PCBs, fertility, industrial chemicals, PAH, animal models, carcinogens, human growth and development, cancer, human exposure, estrogen response, reproductive processes, estrogen receptors, biological effects

Relevant Websites: Exit EPA icon

Progress and Final Reports:

Original Abstract
1999 Progress Report
2000 Progress Report
2001 Progress Report