ANALYSIS OF RECOVERY ACTIVITIES IONOTROPIC GLUTAMATERGIC RECEPTOR MECHANISMS AFTER CRYOPRESERVATION OF BRAIN SLICES

DOI: https://doi.org/None

Mokrushin A.A., Borovikov S.E.

Introduction. We analyzed the hypothesis of cryopreservation of small volumes of nervous tissue (brain slices) without the use of traditional cryoprotectants. The aim of the study. The effects of freezing (-10°C) and rewarming to +37°C examined for changes in activity of glutamatergic ionotropic mechanisms: α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) and N-methyl-D-aspartate (NMDA) receptors. Сhanges in water content in slices were determined after cryopreservation. Methods. Experiments were carried outin slices of the rat olfactory cortex. The viability of slices determined from the change of the AMPA amplitudes and NMDA excitatory postsynaptic components of field potentials have been usedat two speeds of freezing/thawing: slow (0,1°C/min) and rapid (9,0°C/min). Results. The activities of AMPA and NMDA receptor mechanisms recovered at a slow rate of freezing/rewarming, while at the rapid rate – there was noted the hyperactivation of AMPA-dependent mechanisms, NMDA activity was blocked. The water content in slices during thawing was maximal when using a rapid freezing/rewarming cycles. Swelling of slices was negligible at the slow rate of freezing/rewarming. Dipeptide L-carnosine was ineffective as a cryoprotectant for preservation and restoration the activity of NMDA receptors. This peptide prevented the excessive swelling of slices after cryopreservation. Conclusion. Cryopreservation of small volumes of the nervous tissue (brain slices) at a slow rate (0,1°C/min), freezing/rewarming and deep-freezing (-10°C) without the use of cryoprotectants is optimal for cryopreservation and their possible use as grafts.
Keywords: 
brain slices, freezing/rewarming, AMPA, NMDA receptors
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