Research reportGlucose starvation stimulates Zn2+ toxicity in cultures of cerebellar granule neurons

Zinc chloride (0.02 mM, 3 h) 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 ZnCl2 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 Ca2+ uniporter blocker) almost entirely protected CGNs from the toxic effect of ZnCl2 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-3 AM) demonstrated that 1.5 h-exposure to GD induced intensive increase of Fluo-4 fluorescence and small increase of FluoZin-3 fluorescence in neurons. The supplementation of medium with ZnCl2 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 [Zn2+]i increase caused by zinc addition under GD, while ruthenium red did cause significant increase of [Zn2+]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.

► Zinc and calcium ions as synergists in neuronal toxicity under glucose deprivation. ► Glucose ablation potentiates zinc toxicity in neurons. ► Mitochondria are involved in zinc toxicity. ► Zinc ions toxicity is exerted after transport in mitochondria.