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Na+/Ca2+ exchange and regulation of cytoplasmic concentration of calcium in rat cerebellar neurons treated with glutamate

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Abstract

In the present work, the forward and/or reversed Na+/Ca2+ exchange in cerebellar granular cells was suppressed by substitution of Na +0 by Li+ before, during, and after exposure to glutamate for varied time and also using the inhibitor KB-R7943 of the reversed exchange. After glutamate challenge for 1 min, Na +0 /Li+ substitution did not influence the recovery of low [Ca2+]i in a calcium-free medium. A 1-h incubation with 100 μM glutamate induced in the neurons a biphasic and irreve rsible [Ca2+]i rise (delayed calcium deregulation (DCD)), enhancement of [Na+]i, and decrease in the mitochondrial potential. If Na +0 had been substituted by Li+ before the application of glutamate, i.e. the exchange reversal was suppressed during the exposure to glutamate, the number of cells with DCD was nearly fourfold lowered. However, addition of the Na+/K+-ATPase inhibitor ouabain (0.5 mM) not preventing the exchange reversal also decreased DCD in the presence of glutamate. Both exposures decreased the glutamate-caused loss of intracellular ATP. Glucose deprivation partially abolished protective effects of the Na +0 /Li+ substitution and ouabain. KB-R7943 (10 μM) increased 7.4-fold the number of cells with the [Ca2+]i decreased to the basal level after the exposure to glutamate. Thus, reversal of the Na+/Ca2+ exchange reinforced the glutamate-caused perturbations of calcium homeostasis in the neurons and slowed the recovery of the decreased [Ca2+]i in the post-glutamate period. However, for development of DCD, in addition to the exchange reversal, other factors are required, in particular a decrease in the intracellular concentration of ATP.

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Abbreviations

[Ca2+]i, [Na+]i, [Ca2+]0, [Na+]0 :

intracellular and outer concentrations of Ca2+ and Na+

DCD:

delayed clacium deregulation

DNP:

dinitrophenol

KB-R7943:

2-2-[4-(4-nitrobenzyloxyphenyl]ethylisothiourate methanesulfonate

NMDA:

N-methyl-D-aspartate

PM:

plasma membrane

ΔΨm :

mitochondrial membrane potential

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Authors and Affiliations

  1. Research Center for Children’s Health, Russian Academy of Medical Sciences, Lomonosovskii pr. 2/62, 119991, Moscow, Russia

    T. P. Storozhevykh, E. G. Sorokina, A. V. Vabnitz, Ya. E. Senilova, G. R. Tukhbatova & V. G. Pinelis

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  1. T. P. Storozhevykh
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  2. E. G. Sorokina
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  3. A. V. Vabnitz
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  4. Ya. E. Senilova
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  5. G. R. Tukhbatova
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Correspondence to T. P. Storozhevykh.

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Original Russian Text © T. P. Storozhevykh, E. G. Sorokina, A. V. Vabnitz, Ya. E. Senilova, G. R. Tukhbatova, V. G. Pinelis, 2007, published in Biokhimiya, 2007, Vol. 72, No. 7, pp. 923–933.

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Storozhevykh, T.P., Sorokina, E.G., Vabnitz, A.V. et al. Na+/Ca2+ exchange and regulation of cytoplasmic concentration of calcium in rat cerebellar neurons treated with glutamate. Biochemistry Moscow 72, 750–759 (2007). https://doi.org/10.1134/S0006297907070097

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