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Expression profiling of estrogenic compounds using a sheepshead minnow cDNA macroarray.

Patrick Larkin1, Leroy C Folmar, Michael J Hemmer

  • 1Department of Biochemistry and Molecular Biology and Center for Biotechnology, University of Florida, Gainesville, Florida, USA. pmlarkin@ufl.edu

EHP Toxicogenomics : Journal of the National Institute of Environmental Health Sciences
|May 9, 2003
PubMed
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Estrogenic compounds like E2, EE2, DES, and MXC alter fish gene expression similarly. A sheepshead minnow macroarray revealed distinct genetic responses to various environmental estrogens.

Area of Science:

  • Environmental toxicology
  • Molecular biology
  • Ecotoxicology

Background:

  • Anthropogenic compounds can bind to the estrogen receptor (ER) in vertebrates, disrupting homeostasis by altering gene expression.
  • Estrogenic chemicals are known to affect gene expression patterns in aquatic organisms, posing ecological risks.

Purpose of the Study:

  • To characterize the expression of 30 genes in sheepshead minnow (Cyprinodon variegatus) using a cDNA macroarray.
  • To investigate the gene expression profiles induced by various estrogenic compounds in fish models.

Main Methods:

  • Utilized a sheepshead minnow cDNA macroarray to analyze the expression of 30 genes.
  • Exposed fish to known estrogenic compounds including 17 beta-estradiol (E2), 17 alpha-ethinyl estradiol (EE2), diethylstilbestrol (DES), methoxychlor (MXC), p-nonylphenol, and endosulfan.

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  • Employed differential display reverse transcriptase-polymerase chain reaction to identify previously regulated genes.

    • Main Results:

    • 17 beta-estradiol (E2), 17 alpha-ethinyl estradiol (EE2), diethylstilbestrol (DES), and methoxychlor (MXC) exhibited similar gene expression patterns.
    • p-Nonylphenol showed an identical pattern to E2, EE2, DES, and MXC, with the additional upregulation of ubiquitin-conjugating enzyme 9.
    • Endosulfan exposure resulted in gene expression patterns similar to controls, except for the upregulation of estrogen receptor alpha and downregulation of 3-hydroxy-3-methylglutaryl-coenzyme A reductase.
    • The estrogen-responsive cDNA macroarray demonstrated dose-dependent changes in gene expression in response to EE2.

    Conclusions:

    • Estrogenic compounds induce conserved gene expression signatures in sheepshead minnows.
    • The developed cDNA macroarray is a sensitive tool for detecting environmental estrogen effects and dose-dependent responses.
    • Understanding these genetic responses is crucial for assessing the ecological impact of endocrine-disrupting chemicals.