title: Hexabromocyclododecane inhibits depolarization-induced increase in intracellular calcium levels and neurotransmitter release in PC12 cells.
abstract: Environmental levels of the brominated flame retardant (BFR) hexabromocyclododecane (HBCD) have been increasing. HBCD has been shown to cause adverse effects on learning and behavior in mice, as well as on dopamine uptake in rat synaptosomes and synaptic vesicles. For other BFRs, alterations in the intracellular Ca(2+) homeostasis have been observed. Therefore, the aim of this study was to investigate whether the technical HBCD mixture and individual stereoisomers affect the intracellular Ca(2+) concentration ([Ca(2+)](i)) in a neuroendocrine in vitro model (PC12 cells). [Ca(2+)](i) and vesicular catecholamine release were measured using respectively single-cell Fura-2 imaging and amperometry. Exposure of PC12 cells to the technical HBCD mixture or individual stereoisomers did neither affect basal [Ca(2+)](i), nor the frequency of basal neurotransmitter release. However, exposure to HBCD (0-20 microM) did cause a dose-dependent reduction of a subsequent depolarization-evoked increase in [Ca(2+)](i). This effect was apparent only when HBCD was applied at least 5 min before depolarization (maximum effect after 20 min exposure). The effects of alpha- and beta-HBCD were comparable to that of the technical mixture, whereas the inhibitory effect of gamma-HBCD was larger. Using specific blockers of L-, N- or P/Q-type voltage-gated Ca(2+) channels (VGCCs) it was shown that the inhibitory effect of HBCD is not VGCC-specific. Additionally, the number of cells showing depolarization-evoked neurotransmitter release was markedly reduced following HBCD exposure. Summarizing, HBCD inhibits depolarization-evoked [Ca(2+)](i) and neurotransmitter release. As increasing HBCD levels should be anticipated, these findings justify additional efforts to establish an adequate exposure, hazard and risk assessment.
authors.affiliation: Institute for Risk Assessment Sciences (IRAS), Utrecht University, NL-3508 TD Utrecht, The Netherlands. m.dingemans@uu.nl
authors.forename: Milou M L
authors.initials: MM
authors.lastname: Dingemans
authors.forename: Harm J
authors.initials: HJ
authors.lastname: Heusinkveld
authors.forename: Aart
authors.initials: A
authors.lastname: de Groot
authors.forename: Ake
authors.initials: A
authors.lastname: Bergman
authors.forename: Martin
authors.initials: M
authors.lastname: van den Berg
authors.forename: Remco H S
authors.initials: RH
authors.lastname: Westerink
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002395:Catecholamines / Q000378:metabolism; D016764:Cell Polarity / Q000187:drug effects / Q000502:physiology*; D002470:Cell Survival / Q000187:drug effects; D022162:Cytoplasmic Vesicles / Q000187:drug effects / Q000378:metabolism; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D003627:Data Interpretation, Statistical; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D005411:Flame Retardants / Q000633:toxicity*; D005456:Fluorescent Dyes; D016257:Fura-2; D006842:Hydrocarbons, Brominated / Q000633:toxicity*; D018377:Neurotransmitter Agents / Q000378:metabolism*; D016716:PC12 Cells; D051381:Rats; D013237:Stereoisomerism
chemical_list: D002121:Calcium Channel Blockers; D002395:Catecholamines; D005411:Flame Retardants; D005456:Fluorescent Dyes; D006842:Hydrocarbons, Brominated; D018377:Neurotransmitter Agents; C089796:hexabromocyclododecane; D002118:Calcium; D016257:Fura-2