title: Gene expression profiling in zebrafish embryos exposed to diclofenac, an environmental toxicant.
abstract: Pharmaceuticals are continually released in the environment and therefore pollution from drugs is a pressing problem in the environment. Diclofenac, 2-[(2,6-dichlorophenyl)amino]phenylacetic acid is a FDA approved non-steroidal anti-inflammatory drug (NSAID) for the treatment of inflammation. This pharmaceutical has been found as pollutant in superficial waters. Danio rerio (zebrafish) embryo has been used as a model organism for acute pollutant toxicity tests in order to identify morphological alterations in development and death rate. Through the combination of mRNA differential display and quantitative Real Time experiments, we analyzed the alterations of gene expression in zebrafish embryos left to develop in the presence of diclofenac and thereby assess the molecular mechanism involved in ecotoxicity of diclofenac polluted waters. This approach, in embryos exposed to 1.25 mg/l drug for 48 h, allowed identifying 36 different genes, with both known and unknown functions, whose transcription is differentially regulated. The identity and ontological classification of these genes is presented. The wide variety of functional classes of transcripts isolated in this screen reflects the diverse spectrum of influences operating across diclofenac exposure. Of these 36 genes, several have been selected for detailed quantitative Real Time analysis to validate the screen. Our results, for the first time, provide an insight into some of the varied and novel molecular networks following zebrafish exposure to diclofenac polluted waters.
authors.affiliation: Department of Life Sciences, University of Naples II, Via Vivaldi 43, 81100 Caserta, Italy. bruna.defelice@unina2.it
authors.forename: Bruna
authors.initials: B
authors.lastname: De Felice
authors.forename: Luisa
authors.initials: L
authors.lastname: Copia
authors.forename: Marco
authors.initials: M
authors.lastname: Guida
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D019295:Computational Biology; D004008:Diclofenac / Q000633:toxicity*; D004625:Embryo, Nonmammalian / Q000378:metabolism*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D014874:Water Pollutants, Chemical / Q000633:toxicity*; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism*
chemical_list: D014874:Water Pollutants, Chemical; D004008:Diclofenac