Detection of PBDE effects on mRNA expression in chicken (Gallus domesticus) neuronal cells using real-time RT-PCR and a new differential display method.
Polybrominated diphenyl ethers (PBDEs) are used as flame retardants in a wide range of consumer products. Previous studies have suggested that PBDEs can disrupt thyroid hormone homeostasis and the developing central nervous system in rodents, but few studies have determined whether PBDEs cause similar effects in birds. An in vitro method was used to determine effects of a commercial PBDE flame retardant (DE-71) on mRNA expression in primary chicken neuronal cells derived from the cerebral hemisphere. Real-time RT-PCR assays were developed to quantify changes in mRNA abundance of genes associated with the thyroid hormone pathway; thyroid hormone receptors (TRalpha and TRbeta) and transthyretin (TTR). We also used a new differential display PCR methodology, fluorescent RNA arbitrarily primed PCR (FRAP-PCR), to determine additional effects of DE-71 on mRNA expression. Neither of the TRs responded to DE-71 exposure, but TTR mRNA decreased approximately 2-fold following exposure to 0.1, 1 and 3 microM DE-71. Candidate transcripts associated with signal transduction, neurosteroidogenesis, and neurite and axonal growth were up-regulated by DE-71 exposure. Taken together, the findings from this study indicate that this in vitro cell culture method can be used to characterize the effects of PBDEs in the avian brain.
Crump
Doug
D
Environment Canada, National Wildlife Research Centre, 1125 Colonel By Drive, Ottawa, Ontario, Canada K1S 5B6. doug.crump@ec.gc.ca
Jagla
Magdalena M
MM
Chiu
Suzanne
S
Kennedy
Sean W
SW
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D054022:Cerebrum / Q000187:drug effects* / Q000196:embryology; D002642:Chick Embryo; D004305:Dose-Response Relationship, Drug; D005411:Flame Retardants / Q000633:toxicity*; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression / Q000187:drug effects*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D055768:Halogenated Diphenyl Ethers; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D010647:Phenyl Ethers / Q000633:toxicity*; D011075:Polybrominated Biphenyls / Q000633:toxicity*; D011228:Prealbumin / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D037021:Thyroid Hormone Receptors alpha / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D037042:Thyroid Hormone Receptors beta / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects
D005411:Flame Retardants; D055768:Halogenated Diphenyl Ethers; D010647:Phenyl Ethers; D011075:Polybrominated Biphenyls; D011228:Prealbumin; D012333:RNA, Messenger; D037021:Thyroid Hormone Receptors alpha; D037042:Thyroid Hormone Receptors beta; C086401:pentabromodiphenyl ether