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BMC Physiology

, 7:12

First Online: 05 December 2007Received: 05 July 2007Accepted: 05 December 2007

Abstract

BackgroundThe 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.

ResultsThe 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.

ConclusionOverall, 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.

Electronic supplementary materialThe online version of this article doi:10.1186-1472-6793-7-12 contains supplementary material, which is available to authorized users.

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Author: Pål A Olsvik - Kai K Lie - Øystein Sæle - Monica Sanden

Source: https://link.springer.com/







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