title: Immunoquantitation and microsomal monooxygenase activities of hepatic cytochromes P4501A and P4502B and chlorinated hydrocarbon contaminant levels in polar bear (Ursus maritimus).
abstract: Contamination of the Arctic ecosystem by anthropogenic compounds has resulted in exposure of polar bear (Ursus maritimus) to lipophilic chlorinated hydrocarbon contaminants (CHCs) accumulated through the marine food web. Liver samples were collected from 16 adult male polar bears in the Canadian arctic and subjected to chemical analysis for CHCs and metabolites, determination of alkoxyresorufin O-dealkylase activities, and immunoquantitation of cytochrome P450 (CYP) protein levels. We report on the relationships between the hepatic microsomal levels of immunoreactive CYP1A and CYP2B isozymes, catalytic activities, and hepatic CHC and metabolite concentrations in polar bear. We specifically explored the influence of several CHCs on the induction of hepatic CYP in polar bear and the potential use of immunoassay quantitation as a bioindicator of CHC exposure. Polychlorinated biphenyls (PCB) classed as CYP1A and mixed CYP1A/CYP2B inducers accounted for about 25% of the total PCB residues present (18,680 +/- 5053 ng/g lipid). CYP1A protein content correlated strongly with hepatic levels of PCBs, PCDDs (0.032 +/- 0.018 ng/g lipid, and PCDFs (0.011 +/- 0.007 ng/g lipid) and their corresponding toxic equivalents (TEQ, 0.377 +/- 0.182 ng/g lipid). Mono-ortho-CB-156, CB-157, and CB-105 were the predominant TEQ contributors. Correlations between CYP2B protein content and CHC residue levels in polar bear liver suggested that ortho-chlorine-substituted PCBs and chlordanes were the major contributors to CYP2B induction. CYP1A and CYP2B contents were therefore good indicators of CHC exposure in polar bear liver. Ethoxyresorufin, pentoxyresorufin, and benzyloxyresorufin O-dealkylase activities increased with increasing CYP1A protein content up to protein levels of approximately 5 pmol/mg, suggesting that all three activities were primarily CYP1A-mediated. These results were substantiated by antibody inhibition experiments. In summary, immunoquantitated CYP1A and CYP2B isozymes are a more reliable measure of exposure to CHC inducers than alkoxyresorufin O-dealkylase activities in polar bear.
authors.affiliation: Centre for Analytical and Environmental Chemistry, Department of Chemistry, Carleton University, Ottawa, Ontario, Canada.
authors.forename: R J
authors.initials: RJ
authors.lastname: Letcher
authors.forename: R J
authors.initials: RJ
authors.lastname: Norstrom
authors.forename: S
authors.initials: S
authors.lastname: Lin
authors.forename: M A
authors.initials: MA
authors.lastname: Ramsay
authors.forename: S M
authors.initials: SM
authors.lastname: Bandiera
mesh_terms: D000818:Animals; D019362:Cytochrome P-450 CYP2B1; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000378:metabolism*; D004586:Electrophoresis; D004785:Environmental Pollutants / Q000378:metabolism* / Q000633:toxicity; D004790:Enzyme Induction / Q000187:drug effects; D006843:Hydrocarbons, Chlorinated / Q000378:metabolism* / Q000633:toxicity; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000494:pharmacology; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism*; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D010088:Oxidoreductases / Q000037:antagonists & inhibitors / Q000378:metabolism; D010575:Pesticides / Q000378:metabolism; D001503:Ursidae / Q000378:metabolism*
chemical_list: D065607:Cytochrome P-450 Enzyme Inhibitors; D004785:Environmental Pollutants; D006843:Hydrocarbons, Chlorinated; D007074:Immunoglobulin G; D007527:Isoenzymes; D010575:Pesticides; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; D019362:Cytochrome P-450 CYP2B1