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pH Changes in the Mitochondrial Matrix and Cytosol under Glutamate Deregulation of Ca2+ Homeostasis in Cultured Rat Hippocampal Neurons

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Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology Aims and scope

Abstract

The effect of high concentrations of glutamate (Glu) on primary cultures of neurons from the rat brain led to a strong depolarization of mitochondria, which developed synchronously with a secondary increase in the intracellular free Ca2+ concentration (delayed calcium deregulation, DCD). Simultaneously with measurements of the intracellular free Ca2+ concentration ([Ca2+]i), pH was measured in the mitochondrial matrix (pHm) and cytosol (pHc) of neurons when exposed to a toxic dose of Glu (100 µM). For this purpose, pH-sensitive green fluorescent protein mtYFP in mitochondria and pH-sensitive red fluorescent protein mKate in cytosol were expressed in primary cultures from the hippocampus of newborn rats. The resulting neuronal culture was loaded with the Ca2+ indicator Fura-FF; [Ca2+]i, pHm and pHc were simultaneously measured in those neurons that expressed both mtYFP and mKate. It was found that during the first phase of the [Ca2+]i response to Glu, when partial depolarization of mitochondria was observed, there was an increase in the pH gradient between the mitochondrial matrix and the cytosol (ΔpH), which compensated for the decrease in the electrical component of the mitochondrial potential (∆Ψm), thereby maintaining the constancy of the electrochemical potential of mitochondria. The development of DCD led to an abrupt decrease in ∆Ψm and ΔpH in the soma of neurons; however, a complete collapse of ΔpH was not observed. This may mean that DCD was not caused by a nonspecific megapore in the inner mitochondrial membrane (mPTP), as is commonly believed. Alternatively, part of the mitochondria in the soma of neurons could retain the barrier properties of the inner membrane and did not form mPTP even with the development of DCD and reaching a high [Ca2+]i plateau.

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Funding

This work was carried out according to the plans of State tasks of the Ministry of Health of the Russian Federation (project no. AAAAA 19-119012590191-3) and the Ministry of Science and Higher Education of the Russian Federation (project no. FGFU-2022-0012).

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Authors and Affiliations

  1. Institute of General Pathology and Pathophysiology, Russian Academy of Medical Sciences, 125315, Moscow, Russia

    A. M. Surin & R. R. Sharipov

  2. Scientific Centre for Children’s Health, Russian Academy of Medical Sciences, 119991, Moscow, Russia

    A. M. Surin & V. G. Pinelis

  3. Pirogov Russian National Research Medical University, 117513, Moscow, Russia

    L. R. Gorbacheva & I. G. Savinkova

  4. Department of Human and Animal Physiology, School of Biology, Moscow State University, 119234, Moscow, Russia

    L. R. Gorbacheva

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  1. A. M. Surin
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Correspondence to A. M. Surin.

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All experimental procedures on animals were performed in accordance with ethical principles and regulatory documents recommended by the European Scientific Foundation (ESF) and the Declaration on humane treatment of animals and in accordance with the Order of the Ministry of Health and Social Development of Russia no. 708n dated 23.08.2010 “On approval of the rules of laboratory practice”.

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Translated by E. Puchkov

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Surin, A.M., Gorbacheva, L.R., Savinkova, I.G. et al. pH Changes in the Mitochondrial Matrix and Cytosol under Glutamate Deregulation of Ca2+ Homeostasis in Cultured Rat Hippocampal Neurons. Biochem. Moscow Suppl. Ser. A 16, 236–245 (2022). https://doi.org/10.1134/S1990747822040079

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