title: A characterization of the ZFL cell line and primary hepatocytes as in vitro liver cell models for the zebrafish (Danio rerio).
abstract: The zebrafish (Danio rerio) is a widely used model species in biomedical research. The ZFL cell line, established from zebrafish liver, and freshly isolated primary hepatocytes from zebrafish have been used in several toxicological studies. However, no previous report has compared and characterized these two systems at the level of gene expression. The aim of this study was to evaluate the ZFL cell line in comparison to primary hepatocytes as in vitro models for studying effects of environmental contaminants in zebrafish liver. Using quantitative real-time PCR, the basal level and transcriptional induction potential of key genes involved in toxic responses in the ZFL cell line, primary hepatocytes and whole liver from zebrafish were compared. The study showed that the ZFL cells have lower levels of mRNA of most selected genes compared to zebrafish liver. The induced gene transcription following exposure to ligand was much lower in ZFL cells compared to zebrafish primary hepatocytes at the doses tested. Importantly, oestrogen receptor and vitellogenin genes showed low basal transcription and no induction response in the ZFL cell line. In conclusion, it appears that primary hepatocytes are well suited for studying environmental contaminants including xenoestrogens, but may show large sex-dependent differences in gene transcription. The ZFL cell line shows potential in toxicological studies involving the aryl hydrocarbon receptor pathway. However, low potential for transcriptional induction of genes in general should be expected, especially notable when studying estrogenic responses.
authors.affiliation: Department of Biology, University of Bergen, Bergen, Norway. Electronic address: marta.eide@bio.uib.no.
authors.forename: Marta
authors.initials: M
authors.lastname: Eide
authors.affiliation: Department of Molecular Biology, University of Bergen, Bergen, Norway.
authors.forename: Marte
authors.initials: M
authors.lastname: Rusten
authors.affiliation: Department of Molecular Biology, University of Bergen, Bergen, Norway.
authors.forename: Rune
authors.initials: R
authors.lastname: Male
authors.affiliation: Department of Biology, University of Bergen, Bergen, Norway.
authors.forename: Knut Helge Midtbø
authors.initials: KH
authors.lastname: Jensen
authors.affiliation: Department of Biology, University of Bergen, Bergen, Norway.
authors.forename: Anders
authors.initials: A
authors.lastname: Goksøyr
mesh_terms: D000818:Animals; D002460:Cell Line; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology*; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics; D012737:Sex Factors; D014116:Toxicology / Q000379:methods*; D014874:Water Pollutants, Chemical / Q000633:toxicity; D015027:Zebrafish / Q000502:physiology*
chemical_list: D018160:Receptors, Cytoplasmic and Nuclear; D014874:Water Pollutants, Chemical
keywords: Environmental contaminants; Gene transcription; In vitro models; Nuclear receptors; Quantitative real-time PCR; Vitellogenin