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http://hdl.handle.net/1956/2675
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| Tittel: | Spatial transcription of CYP1A in fish liver |
| Forfattere: | Olsvik, Pål A.
Lie, Kai K.
Sæle, Øystein
Sanden, Monica |
| Dato: | 5-Dec-2007 |
| Forlag: | BioMed Central |
| Sitering: | BMC Physiology 2007, 7:12 |
| Sammendrag: | Background: The aim of this work was to study how evenly detoxifying genes are transcribed
spatially in liver tissue of fish. Ten Atlantic salmon Salmo salar were intraperitoneally injected with
50 mg/kg of the strong CYP1A inducer β-naphthoflavone and liver tissue harvested seven days
later. The liver from 10 control and 10 exposed fish were split into eight sections, RNA extracted
and three reference (β-actin, elongation factor 1AB (EF1AB)) and two detoxifying genes (CYP1A
and GST) quantified with real-time RT-PCR. The cellular localization of the EF1AB and CYP1A
mRNA in the liver of control and β-naphthoflavone treated fish was then determined by in situ
hybridization (ISH) using EF1AB and CYP1A biotinylated oligonucleotide probes.
Results: The study shows that genes encoding phase I and phase II conjugating enzymes are
unevenly transcribed in different parts of the liver of Atlantic salmon seven days after a single-dose
of β-naphthoflavone exposure. Transcription of CYP1A and GST was higher in the middle section
of the liver compared to the distal and proximal parts of the organ. The ISH data suggest that
CYP1A transcription happens mainly in hepatocyte cells in the liver, and that hepatocytes in the
vicinity of blood vessels respond stronger to β-naphthoflavone than cells further away from the
blood supply.
Conclusion: Overall, the qRT-PCR and ISH results reported here suggest that gene expression
analysis should be performed on as pure cell populations as possible. If bulk tissue samples are to
be used, one should always check how evenly the target genes are expressed in tissue sections and
organs in every study. |
| URI: | http://dx.doi.org/10.1186/1472-6793-7-12
http://hdl.handle.net/1956/2675 |
| Vises i samlingene: | Department of Biology
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