title: Glucose starvation stimulates Zn2+ toxicity in cultures of cerebellar granule neurons.
abstract: Zinc chloride (0.02 mM, 3h) did not have any influence on the survival of cerebellar granule neurons (CGNs) incubated in balanced salt solution (BSS). However, in the absence of glucose ZnCl(2) caused severe neuronal damage, decreasing cell survival to 12±2%. Either the blockade of ionotropic glutamate NMDA-receptors with MK-801 or APV or supplementation the medium with ruthenium red (mitochondrial Ca(2+) uniporter blocker) almost entirely protected CGNs from the toxic effect of ZnCl(2) during glucose deprivation (GD). However, NBQX (AMPA/kainate glutamate receptor blocker) did not show protective effect. Measurements of intracellular calcium ions concentration using fluorescent probe (Fluo-4 AM) and zinc ions (FluoZin-3AM) demonstrated that 1.5h-exposure to GD induced intensive increase of Fluo-4 fluorescence and small increase of FluoZin-3 fluorescence in neurons. The supplementation of medium with ZnCl(2) caused equal increase of FluoZin-3 fluorescence at both GD and normoglycemia, whereas the potentiation of Fluo-4 fluorescence by zinc was observed only under GD and could be prevented by MK-801. However, neither MK-801 nor NBQX could influence [Zn(2+)](i) increase caused by zinc addition under GD, while ruthenium red did cause significant increase of [Zn(2+)](i). This data implies that zinc ions during GD induce an additional overload of CGNs with calcium ions that get transported through activated NMDA-channel. Zinc and calcium ions accumulate in mitochondria and amplify individual destructive action on these organelles leading to neuronal death.
authors.affiliation: A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, 119992 Moscow, Russia. isaev@genebee.msu.su
authors.forename: Nickolay K
authors.initials: NK
authors.lastname: Isaev
authors.forename: Ekaterina R
authors.initials: ER
authors.lastname: Lozier
authors.forename: Svetlana V
authors.initials: SV
authors.lastname: Novikova
authors.forename: Denis N
authors.initials: DN
authors.lastname: Silachev
authors.forename: Dmitry B
authors.initials: DB
authors.lastname: Zorov
authors.forename: Elena V
authors.initials: EV
authors.lastname: Stelmashook
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects*; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology*; D002712:Chlorides / Q000633:toxicity*; D004396:Coloring Agents / Q000494:pharmacology; D016291:Dizocilpine Maleate / Q000494:pharmacology; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000187:drug effects*; D018696:Neuroprotective Agents / Q000494:pharmacology; D011810:Quinoxalines / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D012430:Ruthenium Red / Q000494:pharmacology; D017967:Zinc Compounds / Q000633:toxicity*
chemical_list: D002712:Chlorides; D004396:Coloring Agents; D018691:Excitatory Amino Acid Antagonists; D018696:Neuroprotective Agents; D011810:Quinoxalines; D017967:Zinc Compounds; D012430:Ruthenium Red; C062865:2,3-dioxo-6-nitro-7-sulfamoylbenzo(f)quinoxaline; D016291:Dizocilpine Maleate; C016837:zinc chloride; D005947:Glucose; D002118:Calcium