Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
A Gill Filament EROD Assay: Development and Application in Environmental Monitoring
Uppsala University, Teknisk-naturvetenskapliga vetenskapsområdet, Faculty of Science and Technology, Biology, Department of Evolutionary Biology, Department of Environmental Toxicology.
2003 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

A gill filament-based assay for the cytochrome P450 1A (CYP1A)-catalysed activity ethoxyresorufin O-deethylase (EROD) was developed in rainbow trout (Oncorhynchus mykiss) and applied to Atlantic salmon (Salmo salar), Arctic charr (Salvelinus alpinus), Atlantic cod (Gadus morhua), saithe (Pollachius virens), and spotted wolffish (Anarhichas minor). Exposure to waterborne β-naphthoflavone (βNF; 10-6 M) induced branchial EROD activity in all species but the spotted wolffish. In rainbow trout exposed to low concentrations of benzo[a]pyrene (BaP; 10-9 M) and the textile dye indigo (10-8 M) the gills responded more rapidly than the liver to BaP, and indigo induced branchial but not hepatic EROD activity.

A CYP1A-dependent BaP adduct formation was shown in gills of fish exposed to waterborne 3H-BaP, i.e. the adduct formation was enhanced by βNF and blocked by ellipticine (CYP1A inhibitor). The predominant location for BaP adducts was the secondary lamellae (most exposed part of the gill filament), whereas the CYP1A enzyme was also present in the primary lamellae of the gill filament. Hence, in addition to the cell-specific expression of CYP1A an important determinant for the localisation of adducts seemed to be the bioavailability of BaP. This idea is supported by the fact that the CYP1A enzyme was induced only in secondary lamellae by BaP (10-7 M) and indigo (10-6 M), whereas it was induced in both primary and secondary lamellae by 3,3´,4,4´,5-pentachlorobiphenyl (10-8 M). Apparently, readily metabolised inducers (BaP and indigo) are biotransformed in the secondary lamellae.

My results show that gill filament EROD activity is a sensitive biomarker of exposure to waterborne dioxin-like pollutants, and that the assay has potential for use in monitoring. Furthermore, the results suggest that readily metabolised dioxin-like compounds absorbed via the gills may undergo first-pass metabolism in the gill cells and therefore remain undetected by monitoring of EROD activity in the liver.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis , 2003. , p. 45
Series
Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1104-232X ; 920
Keywords [en]
Biology, aquatic, benzo[a]pyrene, biomarker, CYP1A, environmental monitoring, EROD, fish, gill, indigo, PCB
Keywords [sv]
Biologi
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:uu:diva-3913ISBN: 91-554-5841-6 (print)OAI: oai:DiVA.org:uu-3913DiVA, id: diva2:163885
Public defence
2004-01-24, Friessalen, EBC, Norbyvägen 14, Uppsala, 12:00
Opponent
Supervisors
Available from: 2003-12-22 Created: 2003-12-22Bibliographically approved
List of papers
1. A gill filament-based EROD assay for monitoring waterborne dioxin-like pollutants in fish.
Open this publication in new window or tab >>A gill filament-based EROD assay for monitoring waterborne dioxin-like pollutants in fish.
2002 In: Environmental Science and Technology, ISSN 0013-936X, Vol. 36, no 15, p. 3340-3344Article in journal (Refereed) Published
Identifiers
urn:nbn:se:uu:diva-91223 (URN)
Available from: 2003-12-22 Created: 2003-12-22Bibliographically approved
2. EROD activity in gills of anadromous and marine fish as a biomarker of dioxin-like pollutants.
Open this publication in new window or tab >>EROD activity in gills of anadromous and marine fish as a biomarker of dioxin-like pollutants.
Show others...
2003 In: Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, Vol. 136, no 3, p. 235-243Article in journal (Refereed) Published
Identifiers
urn:nbn:se:uu:diva-91224 (URN)
Available from: 2003-12-22 Created: 2003-12-22Bibliographically approved
3. Cell-specific CYP1A expression and benzo(a)pyrene adduct formation in gills of rainbow trout (Oncorhynchus mykiss ) following CYP1A induction in the laboratory and in the field
Open this publication in new window or tab >>Cell-specific CYP1A expression and benzo(a)pyrene adduct formation in gills of rainbow trout (Oncorhynchus mykiss ) following CYP1A induction in the laboratory and in the field
2004 (English)In: Environmental Toxicology and Chemistry, ISSN 0730-7268, E-ISSN 1552-8618, Vol. 23, no 4, p. 874-882Article in journal (Refereed) Published
Abstract [en]

The effect of cytochrome P4501A (CYP1A)induction on cell-specific benzo[a]pyrene (BaP) adduct formation was studied in rainbow trout (Oncorhynchus mykiss) gills. Fish preexposed to β-naphthoflavone (βNF) or caged in a polluted river were exposed to waterborne 3H-benzo[a]pyrene (3H-BaP). The 3H-benzo[a]pyrene adducts in the gill filaments were localized by autoradiography and CYP1A protein by immunohistochemistry. Ethoxyresorufin O-deethylase (EROD) activity was measured using a gill filament-based ex vivo assay. Branchial 3H-BaP binding and EROD activity were enhanced by exposure to βNF or to the river water, and completely blocked by the CYP1A inhibitor ellipticine. The predominant sites of adduct formation were in epithelium of the secondary lamellae and in epithelium of the efferent edge of the gill filament. In βNF-exposed fish, the strongest CYP1A immunoreactivity was observed in differentiating cells and in pillar cells. In fish caged in the polluted river, strong CYP1A immunoreactivity was found in most cells in the secondary lamellae, whereas the primary lamellae were almost devoid of immunoreactivity. Our results reveal a discrepancy between the localization of CYP1A protein and BaP adducts in the gill. Consequently, other factors, such as bioavailability of waterborne polycyclic aromatic hydrocarbons (PAHs) to the target cells, are important for the localization of PAH adducts in the gill.

Keywords
Gill, Cytochrome P4501A, Benzo[a]pyrene, Adduct
Identifiers
urn:nbn:se:uu:diva-91225 (URN)10.1897/03-211 (DOI)
Available from: 2003-12-22 Created: 2003-12-22 Last updated: 2017-12-14Bibliographically approved
4. EROD activities in gills and liver and CYP1A localisation in gills of fish exposed to waterborne benzo[a]pyrene, PCB#126 and indigo.
Open this publication in new window or tab >>EROD activities in gills and liver and CYP1A localisation in gills of fish exposed to waterborne benzo[a]pyrene, PCB#126 and indigo.
Manuscript (Other academic)
Identifiers
urn:nbn:se:uu:diva-91226 (URN)
Available from: 2003-12-22 Created: 2003-12-22 Last updated: 2010-01-13Bibliographically approved

Open Access in DiVA

fulltext(1109 kB)3385 downloads
File information
File name FULLTEXT01.pdfFile size 1109 kBChecksum SHA-1
30ea8aa30eb74814c8da92d51f827d8078689795cd4e8fbb7bd3d3f7e81906f7bdcb224b
Type fulltextMimetype application/pdf
Buy this publication >>

By organisation
Department of Environmental Toxicology
Biological Sciences

Search outside of DiVA

GoogleGoogle Scholar
Total: 3385 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

isbn
urn-nbn

Altmetric score

isbn
urn-nbn
Total: 2322 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf