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Development and validation of a 2,000‐gene microarray for the fathead minnow ( Pimephales promelas )
Author(s) -
Larkin Patrick,
Villeneuve Daniel L.,
Knoebl Iris,
Miracle Ann L.,
Carter Barbara J.,
Liu Li,
Denslow Nancy D.,
Ankley Gerald T.
Publication year - 2007
Publication title -
environmental toxicology and chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.1
H-Index - 171
eISSN - 1552-8618
pISSN - 0730-7268
DOI - 10.1897/06-501r.1
Subject(s) - pimephales promelas , minnow , dna microarray , biology , ecotoxicology , microarray , unigene , gene expression , gene , microarray analysis techniques , computational biology , gene expression profiling , gene chip analysis , genetics , toxicology , transcriptome , fish <actinopterygii> , fishery
Gene microarrays provide the field of ecotoxicology new tools to identify mechanisms of action of chemicals and chemical mixtures. Herein we describe the development and application of a 2,000‐gene oligonucleotide microarray for the fathead minnow Pimephales promelas , a species commonly used in ecological risk assessments in North America. The microarrays were developed from various cDNA and subtraction libraries that we constructed. Consistency and reproducibility of the microarrays were documented by examining multiple technical replicates. To test application of the fathead minnow microarrays, gene expression profiles of fish exposed to 17β‐estradiol, a well‐characterized estrogen receptor (ER) agonist, were examined. For these experiments, adult male fathead minnows were exposed for 24 h to waterborne 17β‐estradiol (40 or 100 ng/L) in a flow‐through system, and gene expression in liver samples was characterized. Seventy‐one genes were identified as differentially regulated by estradiol exposure. Examination of the gene ontology designations of these genes revealed patterns consistent with estradiol's expected mechanisms of action and also provided novel insights as to molecular effects of the estrogen. Our studies indicate the feasibility and utility of microarrays as a basis for understanding biological responses to chemical exposure in a model ecotoxicology test species.