Ethoxyresorufin-O-deethylase and porphyrin analysis in chicken embryo hepatocyte cultures with a fluorescence multiwell plate reader.
Rapid and sensitive methods for measuring ethoxyresorufin-O-deethylase (EROD) induction and total intracellular porphyrin accumulation in cultured chicken embryo hepatocytes exposed to polychlorinated biphenyls (PCBs) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) are described. Cells were cultured in 48-well cell culture plates, and EROD assays were carried out directly within the wells. EROD activity and porphyrin concentrations were determined with a computer-operated fluorescence multiwell plate reader. Porphyrin and EROD assays could be carried out on the same plates or on separate plates. The assays allow one to obtain dose-response curves at rates that are approximately 100 times faster than conventional methods.
Kennedy
S W
SW
Environment Canada, Canadian Wildlife Service, National Wildlife Research Centre, Hull, Quebec.
Lorenzen
A
A
James
C A
CA
Collins
B T
BT
D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D002851:Chromatography, High Pressure Liquid; D003201:Computers; D019363:Cytochrome P-450 CYP1A1; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis*; D007700:Kinetics; D008099:Liver / Q000737:chemistry* / Q000166:cytology / Q000201:enzymology; D010088:Oxidoreductases / Q000032:analysis*; D011166:Porphyrins / Q000032:analysis*; D012015:Reference Standards; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000295:instrumentation
D011166:Porphyrins; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; D019363:Cytochrome P-450 CYP1A1