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Optimization of the Acidic–Alkaline Composition of the Incubation Medium for Long-Term and Reversible Cryopreservation of Brain Slices of Nonhibernating Animals

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Abstract

It was found that the activity of the NMDA receptors (bursts of action potentials) was blocked after long-term cryopreservation of brain slices at –10°С. To reactivate the NMDA receptors after cryopreservation/thawing, alterations in extracellular pH (pHo) of the incubation medium were studied. It was found that in the pHo range of 7.2–7.4 a stepwise acidification of the incubation medium down to pHo 6.5 ± 0.2 occurred. As a result, the NMDA receptor current amplitude decreased to 33%. The use of a hybrid buffer system (carbonate, phosphate, and Tris buffer, pHo 7.7) allowed the reactivation of NMDA receptors. This change in pHo was accompanied by an increase in the NMDA amplitude to 84% compared to the initial amplitude (before cryostorage). Thus, the use of the hybrid buffer system and an increase in the pHo level to 7.7 were the best conditions for reactivation of NMDA receptors during thawing after prolonged cryopreservation.

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ACKNOWLEDGMENTS

The author expresses his gratitude to G.P. Smirnova for her help in conducting the experiments and to S.Ye. Borovikov for his technical assistance in the installation and maintenance of the electrophysiological setup.

Funding

The work was supported by Program of Fundamental Research for state academies for 2013–2020 (GP-14, section 65.2).

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  1. Pavlov Institute of Physiology, Russian Academy of Sciences, nab. Makarova 6, 199034, St. Petersburg, Russia

    A. A. Mokrushin

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  1. A. A. Mokrushin
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Correspondence to A. A. Mokrushin.

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Statement on the welfare of animals. All applied international, national, and institutional principals of animal care and use were met during the experiments.

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Translated by A. Deryabina

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Mokrushin, A.A. Optimization of the Acidic–Alkaline Composition of the Incubation Medium for Long-Term and Reversible Cryopreservation of Brain Slices of Nonhibernating Animals. BIOPHYSICS 66, 812–820 (2021). https://doi.org/10.1134/S0006350921050134

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  • DOI: https://doi.org/10.1134/S0006350921050134

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