title: Differential changes in gene expression in rainbow trout hepatocytes exposed to extracts of oil sands process-affected water and the Athabasca River.
abstract: The oil sands region of northern Alberta represents the world's largest reserves of bitumen, and the accelerated pace of industrial extraction activity has raised concern about the possible impacts on the Athabasca River and its tributaries. An ecotoxicogenomic study was undertaken on Oncorhynchus mykiss trout hepatocytes exposed to extracts of water samples near the oil sand development area, as well as to oil sands process-affected water (OSPW) extracts using the quantitative reverse transcriptase polymerase chain reaction technique. The expression of the following genes (mRNA) was monitored to track changes in xenobiotic biotransformation (CYP1A1, CYP3A4, glutathione S-transferase, multi-drug resistance transporter), estrogenicity (estrogen receptor and vitellogenin), oxidative stress (superoxide dismutase and metallothionein) and DNA repair activity (DNA ligase). The extent of DNA-aromatic hydrocarbon adducts was also determined in cells by immuno-staining. A comparative analysis of gene expression between the river/lake and OSPW samples revealed that CYP3A4, metallothioneins, DNA ligase and GST genes, were specifically expressed by OSPW. Cells exposed to OSPW, commercial naphthenic acids, and benzo(a)pyrene showed increased polyaromatic hydrocarbon DNA-adducts, as determined by cell immunofluorescence analysis. Other genes were induced by all types of water samples, although the induction potential was stronger in OSPW most of the time (e.g., VTG gene was expressed nearly 15-fold by surface waters from the lake and river samples but increased to a maximum of 31-fold in OSPW). A multivariate discriminant function analysis revealed that the lake and river water samples were well discriminated from the OSPW. The CYP3A4 gene was the most highly expressed gene in cells exposed to OSPW and responded less to the lake or river water in the Athabasca River area. This study identified a suite of gene targets that responded specifically to OSPW extracts, which could serve as toxicogenomic fingerprints of OSPW contamination.
authors.affiliation: Fluvial Ecosystem Research, Aquatic Ecosystem Protection Division, Water Science and Technology, Environment Canada, 105 McGill Street, Montréal, Quebec, Canada H2Y 2E7. francois.gagne@ec.gc.ca
authors.forename: F
authors.initials: F
authors.lastname: Gagné
authors.forename: M
authors.initials: M
authors.lastname: Douville
authors.forename: C
authors.initials: C
authors.lastname: André
authors.forename: T
authors.initials: T
authors.lastname: Debenest
authors.forename: A
authors.initials: A
authors.lastname: Talbot
authors.forename: J
authors.initials: J
authors.lastname: Sherry
authors.forename: L M
authors.initials: LM
authors.lastname: Hewitt
authors.forename: R A
authors.initials: RA
authors.lastname: Frank
authors.forename: M E
authors.initials: ME
authors.lastname: McMaster
authors.forename: J
authors.initials: J
authors.lastname: Parrott
authors.forename: G
authors.initials: G
authors.lastname: Bickerton
mesh_terms: D000818:Animals; D051544:Cytochrome P-450 CYP3A / Q000235:genetics; D004247:DNA / Q000737:chemistry; D018736:DNA Adducts / Q000032:analysis; D011088:DNA Ligases / Q000235:genetics; D016002:Discriminant Analysis; D004784:Environmental Monitoring; D005618:Fresh Water; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D005982:Glutathione Transferase / Q000235:genetics; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D006838:Hydrocarbons / Q000737:chemistry / Q000633:toxicity*; D007220:Industrial Waste; D008668:Metallothionein / Q000235:genetics; D017686:Oncorhynchus mykiss; D011084:Polycyclic Aromatic Hydrocarbons / Q000032:analysis; D045483:Rivers; D014874:Water Pollutants, Chemical / Q000633:toxicity*
chemical_list: D018736:DNA Adducts; D006838:Hydrocarbons; D007220:Industrial Waste; D011084:Polycyclic Aromatic Hydrocarbons; D014874:Water Pollutants, Chemical; C095911:polycyclic aromatic hydrocarbons-DNA adduct; D004247:DNA; D008668:Metallothionein; D051544:Cytochrome P-450 CYP3A; D005982:Glutathione Transferase; D011088:DNA Ligases