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Extracellular Alkalosis Reduces the Neurotoxicity of Zinc Ions in Cultured Cerebellar Granule Neurons

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Abstract

Zn2+ is known to be important for the normal brain functions. Disruption of zinc homeostasis and zinc-induced neurotoxicity has been shown to play a role in the development of neurodegenerative diseases. In this work, we investigated the effect of extracellular alkalosis on the zinc ions neurotoxicity in the cultured rat cerebellar granule neurons. Zinc chloride (0.03–0.06 mM, 24 h) added to the culture medium of rat cerebellar granule neurons caused the dose-dependent death of these cells. According to ultrastructural morphological features, the process of cell death could be attributed to necrosis, since it was accompanied by swelling of intracellular organelles and disruption of cell membranes against the background of relatively intact nuclear membranes. Neuronal death was associated with an increase in the level of intracellular free zinc. The toxic effect of zinc ions was significantly decreased when ionotropic glutamate NMDA-receptors were blocked by MK-801 or when the extracellular pH was increased from 7.3 to 7.8, due to a decrease in the zinc overload of the cytoplasm of these cells. The presented results demonstrate that NMDA channels are one of the Zn ion entry pathways in the cultured cerebellar granule neurons. Extracellular alkalosis reduces the zinc overload of the cytoplasm and, consequently, promotes the survival of neurons. Probably, zinc’s neurotoxicity is inextricably linked with changes in the intracellular concentration of protons.

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The data of the paper are available upon request from the corresponding author.

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Authors and Affiliations

  1. M.V. Lomonosov Moscow State University, Leninskiye gory, 119234, Moscow, Russia

    Margarita O. Shedenkova, Sergey A. Golyshev & Nickolay K. Isaev

  2. Research Center of Neurology, Moscow, Russia

    Margarita O. Shedenkova, Elena V. Stelmashook, Elizaveta E. Genrikhs & Nickolay K. Isaev

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  1. Margarita O. Shedenkova
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  2. Elena V. Stelmashook
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  4. Elizaveta E. Genrikhs
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  5. Nickolay K. Isaev
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Contributions

N.K. Isaev, E.V. Stelmashook, and E.E. Genrikhs contributed to conceptualization, contributed to data collection and analyzed the data, and wrote the first draft of the manuscript; E.V. Stelmashook, E.E. Genrikhs, M.O. Shedenkova, and S.A. Golyshev performed the experiments and formal analysis and wrote experimental part of the manuscript; N.K. Isaev edited the figures; and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript. N.K. Isaev and E.V. Stelmashook formatted/submitted the paper.

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Correspondence to Nickolay K. Isaev.

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Handling and experimental procedures with animals were carried out in accordance with Council of the European Community directives 86/609/EEC on the use of animals for experimental research. The experimental protocols were approved by the Ethics committee of the Research Center of Neurology (Protocol No. 12–16/19) Moscow, Russia.

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Highlights

• Zn2+ caused the death of neurons and increase in the intracellular free zinc

• MK-801 decreased Zn2+-toxicity and intracellular free zinc

• The Zn2+ toxicity decreased with an increase in extracellular pH from 7.3 to 7.8

• An increase in extracellular pH reduces the zinc overload in the cytoplasm

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Shedenkova, M.O., Stelmashook, E.V., Golyshev, S.A. et al. Extracellular Alkalosis Reduces the Neurotoxicity of Zinc Ions in Cultured Cerebellar Granule Neurons. Biol Trace Elem Res 201, 856–864 (2023). https://doi.org/10.1007/s12011-022-03214-6

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  • DOI: https://doi.org/10.1007/s12011-022-03214-6

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