title: Polybrominated diphenyl ether (PBDE) effects in rat neuronal cultures: 14C-PBDE accumulation, biological effects, and structure-activity relationships.
abstract: Polybrominated diphenyl ethers (PBDEs), widely used as flame-retardants, are now recognized as globally distributed pollutants, and are detected in most environmental and biological samples, including human blood, adipose tissue, and breast milk. Due to their wide use in commercial products and their persistent nature, long-term exposure to PBDEs may pose a human health risk, especially to children. Our previous reports showed that the commercial PBDE mixture, DE-71, affected protein kinase C (PKC) and calcium homeostasis in a similar way to those of a structurally-related polychlorinated biphenyl (PCB) mixture. These intracellular signaling events are associated with neuronal development and learning and memory function. The objectives of the present study were to test whether environmentally relevant PBDE congeners, with different position and number of bromines, affected PKC translocation in cerebellar granule neuronal cultures and compare the potency and efficacy of PBDE congeners with their 14C-accumulation. All the tested PBDE congeners increased 3H-phorbol ester (PDBu) binding, and a significant effect was seen as low as 10 microM. Among the congeners tested, 2,2',4,4'-tetrabromodiphenyl ether (PBDE 47) increased 3H-PDBu binding in a concentration-dependent manner and to a greater extent than other congeners. These effects were seen at concentrations and exposure times where no cytotoxicity was observed. The efficacy of PBDE congeners varied with their structural composition, and the effects seen on 3H-PDBu binding with some PBDE congeners are similar to those of PCB congeners. Cerebellar granule neurons accumulated all three PBDE congeners (PBDEs 47, 99, and 153) following exposure. At the lowest concentration (0.67 microM), about 13-18% of the total dose of 14C-PBDE congeners was accumulated by these neurons. There were distinct differences in the pattern of 14C-PBDE accumulation among the PBDE congeners. The 14C-PBDE accumulation, either represented as percent basis or nanomole basis, was much lower for the 30.69 microM PBDE 99 and 10.69-30.69 microM PBDE 153 than at the lower concentrations, which may be due to low solubility of these congeners. The accumulation pattern with PBDE 47 did not vary with concentration. On a nanomole accumulation basis, PBDEs 47, 99, and 153 accumulation was linear with time. While the nanomole accumulation was linear with concentration for PBDE 47, it is nonlinear for PBDEs 99 and 153. The pattern of PBDE accumulation seems to correlate with the effects on PKC translocation, with regression values of 0.773-0.991. These results indicate that PBDEs affected PKC translocation in neurons in a similar way to those of other organohalogens, some PBDE congeners are equally efficacious as the respective PCB congeners, and PBDE accumulation correlated well with PKC translocation, suggesting a common mode of action for this group of chemicals.
authors.affiliation: Cellular and Molecular Toxicology Branch, Neurotoxicology and Experimental Toxicology Division, NHEERL, ORD, US Environmental Protection Agency, Research Triangle Park, North Carolina 27711, USA. kodavanti.prasada@epa.gov
authors.forename: Prasada Rao S
authors.initials: PR
authors.lastname: Kodavanti
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Ward
authors.forename: Gabriele
authors.initials: G
authors.lastname: Ludewig
authors.forename: Larry W
authors.initials: LW
authors.lastname: Robertson
authors.forename: Linda S
authors.initials: LS
authors.lastname: Birnbaum
mesh_terms: D000818:Animals; D002250:Carbon Radioisotopes; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D002531:Cerebellum / Q000166:cytology; D004305:Dose-Response Relationship, Drug; D004785:Environmental Pollutants / Q000378:metabolism / Q000633:toxicity*; D005411:Flame Retardants / Q000378:metabolism / Q000633:toxicity*; D055768:Halogenated Diphenyl Ethers; D009474:Neurons / Q000187:drug effects* / Q000201:enzymology; D010647:Phenyl Ethers / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity; D010703:Phorbol Esters / Q000378:metabolism; D011075:Polybrominated Biphenyls / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity; D011493:Protein Kinase C / Q000096:biosynthesis; D051381:Rats; D020318:Rats, Long-Evans; D013329:Structure-Activity Relationship
chemical_list: D002250:Carbon Radioisotopes; D004785:Environmental Pollutants; D005411:Flame Retardants; D055768:Halogenated Diphenyl Ethers; D010647:Phenyl Ethers; D010703:Phorbol Esters; D011075:Polybrominated Biphenyls; C086401:pentabromodiphenyl ether; D011493:Protein Kinase C