title: Characterizing the in vitro hepatic biotransformation of the flame retardant BDE 99 by common carp.
abstract: Polybrominated diphenyl ethers (PBDEs) are a class of flame retardant chemicals known to biomagnify in aquatic foodwebs. However, significant biotransformation of some congeners via reductive dehalogenation has been observed during in vivo and in vitro laboratory exposures, particularly in fish models. Little information is available on the enzyme systems responsible for catalyzing this metabolic pathway in fish. This study was undertaken to characterize the biotransformation of one primary BDE congener, 2,2',4,4',5-pentabromodiphenyl ether (BDE-99), using in vitro techniques. Hepatic sub-cellular fractions were first prepared from individual adult common carp (Cyprinus carpio) to examine metabolism in both microsomal and cytosolic sub-cellular fractions. Debromination rates (i.e. BDE-99 biotransformation to BDE-47) were generally higher in the microsomal fraction than in the cytosolic fraction, and some intra-species variability was observed. Further experiments were conducted to determine the biotransformation kinetics and the influence of specific co-factors, inhibitors and competitive substrates on metabolism using pooled carp liver microsomes. The apparent K(m) and V(max) values were 19.4microM and 1120pmolesh(-1)mgprotein(-1), respectively. Iodoacetate (IaC) and the two thyroid hormones, reverse triodothyronine (rT3) and thyroxine (T4), significantly inhibited the debromination of BDE-99 in microsomal sub-cellular fractions with IC(50) values of 2.2microM, 0.83microM, and >1.0microM, respectively. These results support our hypothesis that deiodinase enzymes may be catalyzing the metabolism of PBDEs in fish liver tissues. Further studies are needed to evaluate metabolic activity in other species and tissues that contain these enzymes.
authors.affiliation: Duke University, Nicholas School of the Environment, Integrated Toxicology and Environmental Health Program, Durham, NC, USA.
authors.forename: Pamela D
authors.initials: PD
authors.lastname: Noyes
authors.forename: Shannon M
authors.initials: SM
authors.lastname: Kelly
authors.forename: Carys L
authors.initials: CL
authors.lastname: Mitchelmore
authors.forename: Heather M
authors.initials: HM
authors.lastname: Stapleton
mesh_terms: D000818:Animals; D002347:Carps / Q000378:metabolism*; D005411:Flame Retardants / Q000493:pharmacokinetics* / Q000633:toxicity; D005982:Glutathione Transferase / Q000378:metabolism; D055768:Halogenated Diphenyl Ethers / Q000493:pharmacokinetics* / Q000633:toxicity; D066298:In Vitro Techniques; D007461:Iodoacetates / Q000494:pharmacology; D007700:Kinetics; D016014:Linear Models; D008099:Liver / Q000201:enzymology / Q000378:metabolism*; D008297:Male; D011441:Propylthiouracil / Q000494:pharmacology; D013347:Subcellular Fractions / Q000201:enzymology / Q000378:metabolism*; D013974:Thyroxine / Q000037:antagonists & inhibitors / Q000378:metabolism; D014285:Triiodothyronine, Reverse / Q000037:antagonists & inhibitors / Q000378:metabolism; D014874:Water Pollutants, Chemical / Q000493:pharmacokinetics* / Q000633:toxicity
chemical_list: C477694:2,2',4,4',5-brominated diphenyl ether; D005411:Flame Retardants; D055768:Halogenated Diphenyl Ethers; D007461:Iodoacetates; D014874:Water Pollutants, Chemical; D014285:Triiodothyronine, Reverse; D011441:Propylthiouracil; D005982:Glutathione Transferase; D013974:Thyroxine