Sensitivity of bald eagle (Haliaeetus leucocephalus) hepatocyte cultures to induction of cytochrome P4501A by 2,3,7,8-tetrachlorodibenzo-p-dioxin.
Graded doses of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) were added to primary hepatocyte cultures of bald eagle (Haliaeetus leucocephalus) embryos to determine their sensitivity to induction of cytochrome P4501A (CYP1A) and porphyrin accumulation. No porphyrin accumulation was observed, but both CYP1A catalytic activity (using the ethoxyresorufin-O-deethylase (EROD) assay) and immunodetectable CYP1A were induced by relatively high concentrations of TCDD. Bald eagle hepatocytes were less sensitive to CYP1A induction than hepatocytes from any other avian species that we have studied to date. These in vitro results are in general agreement with recent assessments of field data, which indicate that bald eagles are relatively insensitive to some of the effects of TCDD and related compounds. Preparation of bald eagle hepatocytes was challenging because existing methods did not yield monolayers of cells. Here we describe details of a new method that was successful for bald eagle hepatocytes. This new method is used routinely in our laboratory to prepare hepatocyte cultures from birds for examination of various biochemical responses to environmental contaminants.
Environment Canada, Canadian Wildlife Service, National Wildlife Research Centre, Carleton University, Ottowa, Ontario, Canada K1A 0H3. sean.kennedy@ec.gc.ca
Sean W
SW
Kennedy
Stephanie P
SP
Jones
John E
JE
Elliott
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D019363:Cytochrome P-450 CYP1A1 / Q000096:biosynthesis* / Q000494:pharmacology; D004625:Embryo, Nonmammalian; D004785:Environmental Pollutants / Q000009:adverse effects*; D004790:Enzyme Induction; D022781:Hepatocytes / Q000187:drug effects / Q000502:physiology*; D000072317:Polychlorinated Dibenzodioxins / Q000009:adverse effects*; D011166:Porphyrins / Q000493:pharmacokinetics; D020304:Raptors
D004785:Environmental Pollutants; D000072317:Polychlorinated Dibenzodioxins; D011166:Porphyrins; D019363:Cytochrome P-450 CYP1A1