Department of Biology, University of Bergen, Bergen, Norway.
Department of Molecular Biology, University of Bergen, Bergen, Norway.
Aquat Toxicol. 2014 Feb;147:7-17. doi: 10.1016/j.aquatox.2013.11.023. Epub 2013 Dec 7.
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.
斑马鱼（Danio rerio）是生物医学研究中广泛使用的模式生物。ZFL 细胞系是从斑马鱼肝脏中建立的，而从斑马鱼新鲜分离的原代肝细胞已被用于多项毒理学研究中。然而，之前没有报道比较和描述这两种系统在基因表达水平上的差异。本研究旨在评估 ZFL 细胞系作为研究环境污染物对斑马鱼肝脏影响的体外模型，与原代肝细胞进行比较。通过实时定量 PCR，比较了 ZFL 细胞系、原代肝细胞和斑马鱼肝脏中参与毒性反应的关键基因的基础水平和转录诱导潜力。研究表明，与斑马鱼肝脏相比，ZFL 细胞中大多数选定基因的 mRNA 水平较低。在测试剂量下，ZFL 细胞中受配体诱导的基因转录水平明显低于斑马鱼原代肝细胞。重要的是，雌激素受体和卵黄蛋白原基因在 ZFL 细胞系中基础转录水平低，且无诱导反应。总之，原代肝细胞非常适合研究包括外源性雌激素在内的环境污染物，但在基因转录方面可能表现出较大的性别依赖性差异。ZFL 细胞系在涉及芳香烃受体途径的毒理学研究中具有潜力。然而，预计基因转录诱导的潜力通常较低，特别是在研究雌激素反应时更为明显。
