Toxicological application of primary hepatocyte cell cultures of Atlantic cod (Gadus morhua)--effects of BNF, PCDD and Cd.

摘要: Abstract Fish primary hepatocyte cultures are commonly used for toxicological assessment of contaminants. So far no one has described a protocol on how to use Atlantic cod hepatocytes in bioassays. In this work we describe an experiment which were able isolate intact liver cells from mature individuals. Hepatic cytochrome P450 1A (CYP1A) expression the isolated was evaluated with situ hybridization after intraperitoneal injection strong CYP1A inducer s-naphthoflavone (BNF). Cod further exposed 1,2,3,7,8-polychlorinated dibenzo- p -dioxin (PCDD) and cadmium (Cd). Transcriptional responses 11 genes quantified (CYP1A, metallothionein (MT), aryl hydrocarbon receptor 2 (AhR2), UDP-glucuronosyltransferase (UGT), glutathione S-transferase (GST), vitellogenin B (VTGB), hypoxia-inducible factor 1 (HIF1), heme oxygenase (HO-1), transferrin, peroxidase (GPx) heat shock protein 70 (HSP70)). Immunohistochemisty evaluation clearly showed elevated mRNA BNF-exposed fish. The transcriptional results that PCDD exposure resulted 311-fold up-regulation Cd 1.82-fold increase MT. Unexpectedly, AhR2 UGT levels not significantly up-regulated PCDD-exposed hepatocytes. HO-1 transferrin dose-dependent response exposure. appears act as endocrine-disrupting metal This study demonstrates research.