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Induction patterns of new CYP1 genes in environmentally exposed rainbow trout
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Organism Biology, Environmental Toxicology.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Organism Biology, Environmental Toxicology.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Organism Biology, Environmental Toxicology.
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2010 (English)In: Aquatic Toxicology, ISSN 0166-445X, E-ISSN 1879-1514, Vol. 98, no 4, 311-321 p.Article in journal (Refereed) Published
Abstract [en]

The cytochrome P4501 (CYP1) gene family comprises four subfamilies in fish: CYP1A, CYP1B, CYP1C, and CYP1D. Only two CYP1 genes, CYP1A1 and CYP1A3, are so far known in rainbow trout (Oncorhynchus mykiss). The present study aimed to identify other CYP1 subfamily genes in rainbow trout, to establish methods for quantitative mRNA expression analysis of these genes, and to determine their basal and induced mRNA expression in gills and liver. Another goal was to examine their mRNA expression in environmentally exposed fish. We cloned four new transcripts, denoted rbCYP1B1, rbCYP1C1, rbCYP1C2, and rbCYP1C3. Levels of these and the previously known rbCYP1A transcripts were determined by real-time PCR in unexposed fish, fish exposed to the potent aryl hydrocarbon receptor (AhR) agonist 3,3',4,4',5-pentachlorobiphenyl (PCB126), and fish caged in various waters in the Uppsala region (Sweden). The mRNA expression patterns observed in unexposed rainbow trout (basal levels) were markedly similar to those reported for orthologous genes in other species. All six transcripts were induced by PCB126 in gills and liver, suggesting all genes to be AhR regulated. The caged fish showed clear rbCYP1 induction in gills at all monitoring sites (up to 70-fold the basal level), whereas the liver responses were weak; induction (up to 5-fold) was recorded only at the Uppsala municipal sewage treatment plant outlet. Gill filament EROD activity was induced at all caging sites. Most interestingly, the rbCYP1 gene response patterns in gills differed among caging sites and among subfamilies. The EROD induction seemed to only reflect induction of rbCYP1A transcription. Response patterns of multiple CYP1 genes in gills and liver could provide an improved monitoring strategy. Such patterns could be used to characterize complex mixtures of AhR agonists and antagonists in aquatic environments.

Place, publisher, year, edition, pages
2010. Vol. 98, no 4, 311-321 p.
National Category
Biological Sciences
Research subject
Ecotoxicology
Identifiers
URN: urn:nbn:se:uu:diva-130469DOI: 10.1016/j.aquatox.2010.03.003ISI: 000278982800001PubMedID: 20371123OAI: oai:DiVA.org:uu-130469DiVA: diva2:349725
Available from: 2010-09-08 Created: 2010-09-08 Last updated: 2017-12-12Bibliographically approved
In thesis
1. Basal and Pollutant-induced Expression of CYP1A, 1B and 1C isoforms in Fish: Implications for Biomonitoring
Open this publication in new window or tab >>Basal and Pollutant-induced Expression of CYP1A, 1B and 1C isoforms in Fish: Implications for Biomonitoring
2013 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Aquatic wildlife are exposed to contaminants in their natural habitats, and toxic pollutants may induce toxicity in sensitive target tissues and cells. There is therefore a need to establish biomarkers to determine exposure to certain classes of contaminants and the subsequent biological responses. In the present work the whole suite of cytochrome P450 1 (CYP1) genes expressed in fish were examined with regard to their inducibility and potential use as biomarkers. Complementary DNA of the CYP1A, 1B and 1C transcripts in rainbow trout and three-spined stickleback were cloned and characterized by quantitative mRNA expression analysis. All CYP1 transcripts could be induced by two selected aryl hydrocarbon receptor (AhR) agoinsts (indigo and PCB 126) in both species, suggesting that all genes were regulated by the AhR. CYP1 mRNA expression profiles induced by PCB 126 and indigo varied over time showing that PCB 126 gave rise to a high and persistent induction in gills and liver while induction by indigo was transient in both organs. The uptake and kinetics of 14C-indigo was studied by autoradiography in rainbow trout. A rapid uptake of 14C-indigo from the water and a subsequent elimination in bile and intestinal contents was observed, explaining the transient CYP1 induction. A high accumulation of 14C-indigo in the gills was completely blocked the CYP1 inhibitor ellipticine, suggesting a CYP1-dependent uptake. High dilutions of a sewage treatment plant effluent containing a complex mixture of pharmaceuticals were investigated. The same water sample both induced CYP1A mRNA expression and inhibited catalytic activity of CYP1A. A field study revealed different induction signatures of the CYP1 genes examined at various locations in Uppsala water environments. As concluded by the temporal and spatial responses obtained, the results of this work suggest that the CYP1 gene panel could be used for biomonitoring of environmental contaminants acting on the CYP system. Further field studies will be required to evaluate this possibility.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2013. 52 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 1044
National Category
Other Biological Topics
Identifiers
urn:nbn:se:uu:diva-198658 (URN)978-91-554-8670-9 (ISBN)
Public defence
2013-05-28, Lindahlsalen, Norbyvägen 18A, Uppsala, 09:15 (English)
Supervisors
Available from: 2013-05-06 Created: 2013-04-22 Last updated: 2013-08-30Bibliographically approved

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Jönsson, Maria E.

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