title: Methylmercury-Dependent Increases in Fluo4 Fluorescence in Neonatal Rat Cerebellar Slices Depend on Granule Cell Migrational Stage and GABAA Receptor Modulation.
abstract: Methylmercury (MeHg) disrupts cerebellar function, especially during development. Cerebellar granule cells (CGC), which are particularly susceptible to MeHg by unknown mechanisms, migrate during this process. Transient changes in intracellular Ca(2+) (Ca(2+) i) are crucial to proper migration, and MeHg is well known to disrupt CGC Ca(2+) i regulation. Acutely prepared slices of neonatal rat cerebellum in conjunction with confocal microscopy and fluo4 epifluorescence were used to track changes induced by MeHg in CGC Ca(2+) i regulation in the external (EGL) and internal granule cell layers (IGL) as well as the molecular layer (ML). MeHg caused no cytotoxicity but did cause a time-dependent increase in fluo4 fluorescence that depended on the stage of CGC development. CGCs in the EGL were most susceptible to MeHg-induced increases in fluo4 fluorescence. MeHg increased fluorescence in CGC processes but only diffusely; Purkinje cells rarely fluoresced in these slices. Neither muscimol nor bicuculline alone altered baseline fluo4 fluorescence in any CGC layer, but each delayed the onset and reduced the magnitude of effect of MeHg on fluo4 fluorescence in the EGL and ML. In the IGL, both muscimol and bicuculline delayed the onset of MeHg-induced increases in fluo4 fluorescence but did not affect fluorescence magnitude. Thus, acute exposure to MeHg causes developmental stage-dependent increases in Ca(2+) i in CGCs. Effects are most prominent in CGCs during development or early stages of migration. GABAA receptors participate in an as yet unclear manner to MeHg-induced Ca(2+) i dysregulation of CGCs.
authors.affiliation: Department of Pharmacology and Toxicology (W.D.A.), Department of Biochemistry and Molecular Biology (A.A.B.), Institute for Integrative Toxicology (A.A.B., W.D.A.), and Neuroscience Program (J.D.M.), Michigan State University, East Lansing, Michigan.
authors.forename: Aaron B
authors.initials: AB
authors.lastname: Bradford
authors.affiliation: Department of Pharmacology and Toxicology (W.D.A.), Department of Biochemistry and Molecular Biology (A.A.B.), Institute for Integrative Toxicology (A.A.B., W.D.A.), and Neuroscience Program (J.D.M.), Michigan State University, East Lansing, Michigan.
authors.forename: Jayme D
authors.initials: JD
authors.lastname: Mancini
authors.affiliation: Department of Pharmacology and Toxicology (W.D.A.), Department of Biochemistry and Molecular Biology (A.A.B.), Institute for Integrative Toxicology (A.A.B., W.D.A.), and Neuroscience Program (J.D.M.), Michigan State University, East Lansing, Michigan atchiso1@msu.edu.
authors.forename: William D
authors.initials: WD
authors.lastname: Atchison
mesh_terms: D000814:Aniline Compounds; D000818:Animals; D000831:Animals, Newborn; D001640:Bicuculline / Q000494:pharmacology; D020013:Calcium Signaling / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002531:Cerebellum / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D003594:Cytoplasmic Granules / Q000187:drug effects / Q000378:metabolism; D005260:Female; D005456:Fluorescent Dyes; D018755:GABA Agonists / Q000494:pharmacology; D018756:GABA Antagonists / Q000494:pharmacology; D066298:In Vitro Techniques; D008297:Male; D008767:Methylmercury Compounds / Q000494:pharmacology*; D009118:Muscimol / Q000494:pharmacology; D011247:Pregnancy; D011689:Purkinje Cells / Q000187:drug effects; D051381:Rats; D011963:Receptors, GABA-A / Q000187:drug effects*; D014966:Xanthenes
chemical_list: D000814:Aniline Compounds; C409648:Fluo 4; D005456:Fluorescent Dyes; D018755:GABA Agonists; D018756:GABA Antagonists; D008767:Methylmercury Compounds; D011963:Receptors, GABA-A; D014966:Xanthenes; D009118:Muscimol; D001640:Bicuculline