A Gill Filament EROD Assay : Development and Application in Environmental Monitoring
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...
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Uppsala universitet, Ekotoxikologi
2003
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ftuppsalauniv:oai:DiVA.org:uu-3913 2023-05-15T14:30:14+02:00 A Gill Filament EROD Assay : Development and Application in Environmental Monitoring Jönsson, Maria 2003 application/pdf http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-3913 eng eng Uppsala universitet, Ekotoxikologi Uppsala : Acta Universitatis Upsaliensis Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, 1104-232X 920 http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-3913 urn:isbn:91-554-5841-6 info:eu-repo/semantics/openAccess Biology aquatic benzo[a]pyrene biomarker CYP1A environmental monitoring EROD fish gill indigo PCB Biologi Doctoral thesis, comprehensive summary info:eu-repo/semantics/doctoralThesis text 2003 ftuppsalauniv 2023-02-23T21:44:16Z 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. Doctoral or Postdoctoral Thesis Arctic charr Arctic atlantic cod Atlantic salmon Gadus morhua Salmo salar Salvelinus alpinus Uppsala University: Publications (DiVA) Arctic |
institution |
Open Polar |
collection |
Uppsala University: Publications (DiVA) |
op_collection_id |
ftuppsalauniv |
language |
English |
topic |
Biology aquatic benzo[a]pyrene biomarker CYP1A environmental monitoring EROD fish gill indigo PCB Biologi |
spellingShingle |
Biology aquatic benzo[a]pyrene biomarker CYP1A environmental monitoring EROD fish gill indigo PCB Biologi Jönsson, Maria A Gill Filament EROD Assay : Development and Application in Environmental Monitoring |
topic_facet |
Biology aquatic benzo[a]pyrene biomarker CYP1A environmental monitoring EROD fish gill indigo PCB Biologi |
description |
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. |
format |
Doctoral or Postdoctoral Thesis |
author |
Jönsson, Maria |
author_facet |
Jönsson, Maria |
author_sort |
Jönsson, Maria |
title |
A Gill Filament EROD Assay : Development and Application in Environmental Monitoring |
title_short |
A Gill Filament EROD Assay : Development and Application in Environmental Monitoring |
title_full |
A Gill Filament EROD Assay : Development and Application in Environmental Monitoring |
title_fullStr |
A Gill Filament EROD Assay : Development and Application in Environmental Monitoring |
title_full_unstemmed |
A Gill Filament EROD Assay : Development and Application in Environmental Monitoring |
title_sort |
gill filament erod assay : development and application in environmental monitoring |
publisher |
Uppsala universitet, Ekotoxikologi |
publishDate |
2003 |
url |
http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-3913 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic charr Arctic atlantic cod Atlantic salmon Gadus morhua Salmo salar Salvelinus alpinus |
genre_facet |
Arctic charr Arctic atlantic cod Atlantic salmon Gadus morhua Salmo salar Salvelinus alpinus |
op_relation |
Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, 1104-232X 920 http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-3913 urn:isbn:91-554-5841-6 |
op_rights |
info:eu-repo/semantics/openAccess |
_version_ |
1766304110360920064 |