title: Delayed effects of methylmercury on the mitochondria of dopaminergic neurons and developmental toxicity in zebrafish larvae (Danio rerio).
abstract: Methylmercury (MeHg) is a known neurotoxicant affecting the central nervous system but effects on dopaminergic (DA) neurons are not well understood. Wild-type zebrafish (Danio rerio) and two transgenic lines: Tg(dat:eGFP) expressing enhanced green fluorescent protein (eGFP) in DA neuron clusters and Tg(dat:tom20 MLS-mCherry) expressing red fluorescence (mCherry) targeted to mitochondria of DA neurons were used to evaluate the effects of micromolar MeHg exposure on DA neuron and whole animal motor function during early development. Three-day-old larvae were exposed to micromolar concentrations of MeHg (0.03, 0.06, and 0.3μM) in system water. Exposure to 0.3μM MeHg caused mortality and significant morphological abnormalities including edema, curvature of the spine, and hemorrhages in zebrafish larvae after a 48h exposure period. At 0.06μM MeHg, the appearance of morphological abnormalities was delayed for 72h and far less severe, whereas 0.03μM MeHg did not cause any morphological defects or mortalities. A delayed but significant reduction in locomotor ability and mCherry fluorescence in specific brain regions in the 0.06μM MeHg exposed larvae suggests that DA neuron function rather than neuron numbers was compromised. Double immunolabeling with tyrosine hydroxylase and pan neural staining showed no effect of MeHg exposure. We have established Tg(dat:tom20 MLS-mCherry) zebrafish larvae as a model which can be used to assess MeHg neurotoxicity and that exposure to low dose MeHg (0.06μM) during development may predispose DA neurons to impairment caused by changes in mitochondrial dynamics.
authors.affiliation: Centre for Advanced Research in Environmental Genomics, University of Ottawa, Ontario K1N 6N5, Canada.
authors.forename: Susie S Y
authors.initials: SS
authors.lastname: Huang
authors.affiliation: Centre for Advanced Research in Environmental Genomics, University of Ottawa, Ontario K1N 6N5, Canada.
authors.forename: Sandra
authors.initials: S
authors.lastname: Noble
authors.affiliation: Centre for Advanced Research in Environmental Genomics, University of Ottawa, Ontario K1N 6N5, Canada.
authors.forename: Rafael
authors.initials: R
authors.lastname: Godoy
authors.affiliation: Centre for Advanced Research in Environmental Genomics, University of Ottawa, Ontario K1N 6N5, Canada.
authors.forename: Marc
authors.initials: M
authors.lastname: Ekker
authors.affiliation: Centre for Advanced Research in Environmental Genomics, University of Ottawa, Ontario K1N 6N5, Canada. Electronic address: laurie.chan@uottawa.ca.
authors.forename: Hing Man
authors.initials: HM
authors.lastname: Chan
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001522:Behavior, Animal / Q000187:drug effects; D059290:Dopaminergic Neurons / Q000187:drug effects* / Q000378:metabolism; D007814:Larva / Q000187:drug effects*; D008767:Methylmercury Compounds / Q000633:toxicity*; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D013550:Swimming; D014874:Water Pollutants, Chemical / Q000633:toxicity*; D015027:Zebrafish / Q000502:physiology*
chemical_list: D008767:Methylmercury Compounds; D014874:Water Pollutants, Chemical
keywords: Delayed response; Dopaminergic neuron; Low dose exposure; Methylmercury; Mitochondria