Development and validation of a 2,000‐gene microarray for the fathead minnow (Pimephales promelas)

Abstract

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 17beta-estradiol, a well-characterized estrogen receptor (ER) agonist, were examined. For these experiments, adult male fathead minnows were exposed for 24 h to waterborne 17beta-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.

The effects of silver nanoparticles on fathead minnow (Pimephales promelas) embryos

“…First, it is a well-characterized environmental model and the species is native to the US. Second, the availability of extensive genetic information on this fish lends to its efficient use in detailed molecular characterization of subcellular effects using available microarrays (Ankley et al 2001;Larkin et al 2007;Kane et al 2008) or other quantitative genetic molecular techniques. Finally, it is the preferred environmental model of the USEPA for use in toxicity studies and thus our knowledge of the biology of this animal will continue to increase through time.…”

Investigations of transcript expression in fathead minnow (Pimephales promelas) brain tissue reveal toxicological impacts of RDX exposure

“…Fathead minnow was selected for this investigation due to its status as the predominant fish model for investigating the acute and chronic toxicity of chemical contaminants (Ankley and Villeneuve, 2006;US EPA, 2002a,b) and its emergence as a robust model for conducting toxicogenomic investigations (Garcia-Reyero et al, 2009a,b, 2008Larkin et al, 2007;Ankley and Villeneuve, 2006;Wintz et al, 2006). Our study incorporated the development of a brain-tissue based de novo cDNA library for fathead minnow enriched for transcripts differentially expressed in response to the MCs, RDX and TNT.…”

Identification of copper-responsive genes in an early life stage of the fathead minnow Pimephales promelas

“…In our study, differential display (Liang and Pardee 1992) was used to identify candidate genes that showed a response to copper. Differential display is a technique that can be used without prior knowledge of an organism's genome and hence was ideal for working with the partially characterized genome of the fathead minnow (Wintz et al 2006;Larkin et al 2007). The purpose of this study was to characterize copper-induced transcriptional changes in fish larvae and to compare this expression profile to other stress studies in fish.…”