title: Effects of exposure to Prestige-like heavy fuel oil and to perfluorooctane sulfonate on conventional biomarkers and target gene transcription in the thicklip grey mullet Chelon labrosus.
abstract: Thicklip grey mullets Chelon labrosus inhabit coastal and estuarine areas where they can be chronically exposed to commonly released pollutants such as polycyclic aromatic hydrocarbons (PAHs) and perfluorinated compounds. These pollutants can also originate from accidental spills, such as the Prestige oil spill in 2002, which resulted in the release of a heavy fuel oil that affected coastal ecosystems in the Bay of Biscay. Peroxisome proliferation (PP), induced biotransformation metabolism, immunosuppression and endocrine disruption are some of the possible biological effects caused by such chemicals. With the aim of studying the effects of organic toxic chemicals on such biological processes at the transcriptional and at the cell/tissue level, juvenile mullets were exposed to the typical mammalian peroxisome proliferator perfluorooctane sulfonate (PFOS), and to fresh (F) and weathered (WF) Prestige-like heavy fuel oil for 2 and 16 days. First, fragments of genes relevant to biotransformation, immune/inflammatory and endocrine disruption processes were cloned using degenerate primers. Fuel oil elicited a significant PP response as proved by the transcriptional upregulation of palmitoyl-CoA oxidase (aox1), peroxisome proliferator activated receptor alpha (pparalpha) and retinoic X receptor, by the AOX1 activity induction and by the increased peroxisomal volume density. PFOS only elicited a significant induction of AOX1 activity at day 2 and of PPARalpha mRNA expression at day 16. All treatments significantly increased catalase mRNA expression at day 16 in liver and at day 2 in gill. Cyp1a transcription (liver and gill) and EROD activity were induced in fuel oil treated organisms. In the case of phase II metabolism only hepatic glutathione S-transferase mRNA was overexpressed in mullets exposed to WF for 16 days. Functionally, this response was reflected in a significant accumulation of bile PAH metabolites. WF treated fish accumulated mainly high molecular weight metabolites while F exposure resulted in accumulation of mainly low molecular ones. Fuel oil significantly regulated immune response related complement component C3 and hepcidin transcription followed by a significant regulation of inflammatory response related apolipoprotein-A1 and fatty acid binding protein mRNAs at day 16. These responses were accompanied by a significant hepatic inflammatory response with lymphocyte accumulations (IRLA) and accumulation of melanomacrophage centers (MMC). PFOS did not elicit any transcriptional response in the studied biotransformation and immune related genes, although histologically significant effects were recorded in IRLA and MMC. A significant reduction of lysosomal membrane stability was observed in all exposed animals. No endocrine disruption effects were observed in liver while brain aromatase mRNA was overexpressed after all treatments at day 2 and estrogen receptor alpha was downregulated under WF exposure at day 16. These results show new molecular and cellular biomarkers of exposure to organic chemicals and demonstrate that in mullets PP could be regulated through molecular mechanisms similar to those in rodents, although the typical mammalian peroxisome proliferator PFOS and heavy fuel oil follow divergent mechanisms of action.
authors.affiliation: Laboratory of Cell Biology and Histology; Department of Zoology and Animal Cell Biology, School of Science and Technology, University of the Basque Country, E-48080 Bilbao, Basque Country, Spain.
authors.forename: E
authors.initials: E
authors.lastname: Bilbao
authors.affiliation: Laboratory of Cell Biology and Histology; Department of Zoology and Animal Cell Biology, School of Science and Technology, University of the Basque Country, E-48080 Bilbao, Basque Country, Spain.
authors.forename: D
authors.initials: D
authors.lastname: Raingeard
authors.affiliation: Laboratory of Cell Biology and Histology; Department of Zoology and Animal Cell Biology, School of Science and Technology, University of the Basque Country, E-48080 Bilbao, Basque Country, Spain.
authors.forename: O Diaz
authors.initials: OD
authors.lastname: de Cerio
authors.affiliation: Laboratory of Cell Biology and Histology; Department of Zoology and Animal Cell Biology, School of Science and Technology, University of the Basque Country, E-48080 Bilbao, Basque Country, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Ortiz-Zarragoitia
authors.affiliation: Laboratory of Cell Biology and Histology; Department of Zoology and Animal Cell Biology, School of Science and Technology, University of the Basque Country, E-48080 Bilbao, Basque Country, Spain.
authors.forename: P
authors.initials: P
authors.lastname: Ruiz
authors.affiliation: Laboratory of Cell Biology and Histology; Department of Zoology and Animal Cell Biology, School of Science and Technology, University of the Basque Country, E-48080 Bilbao, Basque Country, Spain.
authors.forename: U
authors.initials: U
authors.lastname: Izagirre
authors.affiliation: Laboratory of Cell Biology and Histology; Department of Zoology and Animal Cell Biology, School of Science and Technology, University of the Basque Country, E-48080 Bilbao, Basque Country, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Orbea
authors.affiliation: Laboratory of Cell Biology and Histology; Department of Zoology and Animal Cell Biology, School of Science and Technology, University of the Basque Country, E-48080 Bilbao, Basque Country, Spain.
authors.forename: I
authors.initials: I
authors.lastname: Marigómez
authors.affiliation: Laboratory of Cell Biology and Histology; Department of Zoology and Animal Cell Biology, School of Science and Technology, University of the Basque Country, E-48080 Bilbao, Basque Country, Spain.
authors.forename: M P
authors.initials: MP
authors.lastname: Cajaraville
authors.affiliation: Laboratory of Cell Biology and Histology; Department of Zoology and Animal Cell Biology, School of Science and Technology, University of the Basque Country, E-48080 Bilbao, Basque Country, Spain. Electronic address: ibon.cancio@ehu.es.
authors.forename: I
authors.initials: I
authors.lastname: Cancio
mesh_terms: D017738:Alkanesulfonic Acids / Q000633:toxicity*; D000818:Animals; D001483:Base Sequence; D015415:Biomarkers / Q000378:metabolism*; D001711:Biotransformation; D052244:Endocrine Disruptors / Q000633:toxicity; D004781:Environmental Exposure / Q000009:adverse effects; D005466:Fluorocarbons / Q000633:toxicity*; D005786:Gene Expression Regulation / Q000187:drug effects*; D007165:Immunosuppression Therapy; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism; D020675:Peroxisomes / Q000187:drug effects / Q000378:metabolism; D010578:Petroleum / Q000633:toxicity*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D023701:Smegmamorpha / Q000235:genetics* / Q000378:metabolism*; D013030:Spain; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects* / Q000235:genetics
chemical_list: D017738:Alkanesulfonic Acids; D015415:Biomarkers; D052244:Endocrine Disruptors; D005466:Fluorocarbons; D010578:Petroleum; D012333:RNA, Messenger; C076994:perfluorooctane sulfonic acid