Development of the GENIPOL European Flounder (Platichthys flesus) Microarray and Determination of Temporal Transcriptional Responses to Cadmium at Low Dose

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 intraperit...

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Bibliographic Details
Published in:	Environmental science & technology Vol. 40; no. 20; pp. 6479 - 6488
Main Authors:	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
Format:	Journal Article
Language:	English
Published:	Washington, DC American Chemical Society 15-10-2006
Subjects:	Agnatha. Pisces Animal, plant and microbial ecology Animals Applied ecology Arrays Biological and medical sciences Cadmium Cadmium Chloride - administration & dosage Cadmium Chloride - pharmacology Cell cycle Cloning Deoxyribonucleic acid DNA Dose-Response Relationship, Drug Ecotoxicology, biological effects of pollution Environmental Monitoring - methods Flounder Flounder - genetics Flounder - growth & development Fundamental and applied biological sciences. Psychology Gene expression Gene Expression Profiling Gene Expression Regulation, Developmental - drug effects General aspects Genomics Liver Liver - drug effects Liver - growth & development Liver - metabolism Metals Oligonucleotide Array Sequence Analysis - methods Platichthys flesus Pollutants Reverse Transcriptase Polymerase Chain Reaction Transcription, Genetic - drug effects Vertebrates: general zoology, morphology, phylogeny, systematics, cytogenetics, geographical distribution Europe Freshwater environment Marine environment Aquatic environment Cadmium Vertebrata Pisces Low dose Platichthys flesus Heavy metal
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Summary:	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.
Bibliography:	istex:794DADE34298EFE535A87200F3235ACA79F7250F ark:/67375/TPS-78HJV802-R ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0013-936X 1520-5851
DOI:	10.1021/es061142h