Development of the GENIPOL European flounder (Platichthys flesus) microarray and determination of temporal transcriptional responses to cadmium at low dose

TitleDevelopment of the GENIPOL European flounder (Platichthys flesus) microarray and determination of temporal transcriptional responses to cadmium at low dose
Publication TypeJournal Article
Year of Publication2006
AuthorsWilliams, TD, Diab, AM, George, SG, Godfrey, RE, Sabine, V, Conesa, A, Minchin, SD, Watts, PC, Chipman, JK
JournalEnviron Sci Technol
Volume40
Pagination6479-88
KeywordsAnimals Cadmium Chloride/administration & dosage/*pharmacology Dose-Response Relationship; Developmental/drug effects Liver/drug effects/growth & development/metabolism Oligonucleotide Array Sequence Analysis/*methods Reverse Transcriptase Polymerase Chain Reaction Transcription; Drug Environmental Monitoring/methods Flounder/*genetics/growth & development Gene Expression Profiling Gene Expression Regulation; Genetic/*drug effects
Abstract

We have constructed a high density, 13 270-clone cDNA array for the sentinel fish species European flounder (Platichthys flesus), combining clones from suppressive subtractive hybridization and a liver cDNA library; DNA sequences of 5211 clones were determined. Fish were treated by single intraperitoneal injection with 50 micrograms cadmium chloride per kilogram body weight, a dose relevant to environmental exposures, and hepatic gene expression changes were determined at 1, 2, 4, 8, and 16 days postinjection in comparison to saline-treated controls. Gene expression responses were confirmed by real-time reverse transcription polymerase chain reaction (RT-PCR). Blast2GO gene ontology analysis highlighted a general induction of the unfolded protein response, response to oxidative stress, protein synthesis, transport, and degradation pathways, while apoptosis, cell cycle, cytoskeleton, and cytokine genes were also affected. Transcript levels of cytochrome P450 1A (CYP1A) were repressed and vitellogenin altered, real-time PCR showed induction of metallothionein. We thus describe the establishment of a useful resource for ecotoxicogenomics and the determination of the temporal molecular responses to cadmium, a prototypical heavy metal pollutant.

Notes

Williams, Tim D Diab, Amer M George, Stephen G Godfrey, Rita E Sabine, Victoria Conesa, Ana Minchin, Steven D Watts, Phil C Chipman, James K Research Support, Non-U.S. Gov’t United States Environmental science & technology Environ Sci Technol. 2006 Oct 15;40(20):6479-88.

URLhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17120584