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Transcriptional effects of PFOS in isolated hepatocytes from Atlantic salmon Salmo salar L
Anne Vatland Krøvel1, Liv Søfteland, Bente Torstensen
1National Institute of Nutrition and Seafood Research, N-5817 Bergen, Norway.
Abstract:
The main aim of the current in vitro experiment was to search for makers for PFOS exposure in isolated hepatocytes from Atlantic salmon Salmo salar, based on genes responding to PFOS exposure in other animals. Primary cell cultures of hepatocytes were exposed to four concentrations of PFOS (2.1-6.2-15.1-25.0 mg/L) for 24 and 48 h and the transcriptional levels of 12 genes encoding proteins known to respond to PFOS were quantified with real-time RT-PCR. The 12 examined genes were caspase 3B (apoptosis), GSH-Px and HSP70 (cellular stress), CYP1A, CYP3A, GST and UGT (P450 and phase II enzymes), acyl-CoA oxidase, PPARalpha, PPARbeta and PPARgamma (lipid metabolism) and Na(+)-K(+)-ATPase (ion regulation). Most of the studied genes responded in a dose-dependent manner to PFOS exposure, although the transcriptional differences in general where small with regard to fold change. Our results clearly suggest that PFOS exposure enhanced cellular stress in the examined cells, even though the exact mechanisms behind this stress remain unknown. The results from this in vitro experiment showed that genes known to be affected by PFOS exposure in other species also were induced in hepatocytes of Atlantic salmon, giving us the rationale to expand to testing the actual in vivo magnitude of effect in Atlantic salmon exposed to PFOS at doses usually seen in nature/diets.
Insights
Per- and polyfluoroalkyl substances (PFAS) exposure, specifically PFOS, induced cellular stress in Atlantic salmon hepatocytes. Gene expression analysis revealed dose-dependent responses, supporting further in vivo studies.
Area of Science:
- Environmental Toxicology
- Aquatic Toxicology
- Molecular Biology
Background:
- Perfluorooctanesulfonic acid (PFOS) is a persistent environmental contaminant.
- Understanding PFOS effects on aquatic organisms like Atlantic salmon is crucial for ecological risk assessment.
- Gene expression analysis can serve as a sensitive biomarker for chemical exposure.
Purpose of the Study:
- To identify potential biomarkers for PFOS exposure in Atlantic salmon hepatocytes.
- To investigate the transcriptional response of specific genes to PFOS in vitro.
- To assess the dose-dependent effects of PFOS on gene expression in salmon cells.
Main Methods:
- Primary Atlantic salmon hepatocytes were cultured in vitro.
- Cells were exposed to four concentrations of PFOS (2.1-25.0 mg/L) for 24 and 48 hours.
- Real-time RT-PCR was used to quantify the transcriptional levels of 12 genes involved in apoptosis, cellular stress, P450 and phase II metabolism, lipid metabolism, and ion regulation.
Main Results:
- Most of the 12 examined genes showed a dose-dependent response to PFOS exposure.
- PFOS exposure significantly enhanced cellular stress markers in salmon hepatocytes.
- Transcriptional changes were observed, although fold changes were generally small.
Conclusions:
- Genes known to respond to PFOS in other species are also induced in Atlantic salmon hepatocytes.
- PFOS exposure induces cellular stress in Atlantic salmon, suggesting potential toxicological effects.
- These in vitro findings provide a basis for future in vivo studies to assess PFOS effects at environmentally relevant concentrations.
