Abstract
Ischemic postconditioning (PostC) is known to reduce cerebral ischemia/reperfusion (I/R) injury; however, whether the opening of mitochondrial ATP-dependent potassium (mito-KATP) channels and mitochondrial permeability transition pore (mPTP) cause the depolarization of the mitochondrial membrane that remains unknown. We examined the involvement of the mito-KATP channel and the mPTP in the PostC mechanism. Ischemic PostC consisted of three cycles of 15 s reperfusion and 15 s re-ischemia, and was started 30 s after the 7.5 min ischemic load. We recorded N-methyl-d-aspartate receptors (NMDAR)-mediated currents and measured cytosolic Ca2+ concentrations, and mitochondrial membrane potentials in mouse hippocampal pyramidal neurons. Both ischemic PostC and the application of a mito-KATP channel opener, diazoxide, reduced NMDAR-mediated currents, and suppressed cytosolic Ca2+ elevations during the early reperfusion period. An mPTP blocker, cyclosporine A, abolished the reducing effect of PostC on NMDAR currents. Furthermore, both ischemic PostC and the application of diazoxide potentiated the depolarization of the mitochondrial membrane potential. These results indicate that ischemic PostC suppresses Ca2+ influx into the cytoplasm by reducing NMDAR-mediated currents through mPTP opening. The present study suggests that depolarization of the mitochondrial membrane potential by opening of the mito-KATP channel is essential to the mechanism of PostC in neuroprotection against anoxic injury.
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This study was supported by JSPS KAKENHI Grant Number JP16K10735.
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Conception and design or analysis and interpretation of data, or both; YM, IN, YO. Drafting of the manuscript or revising it critically for important intellectual content; YM, IN, YO, SY, TF, YS, HN. Final approval of the manuscript submitted; IN, YS, HN.
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10571_2020_996_MOESM1_ESM.tif
(TIF 126158 kb) Supplementary Fig. 1 Representative microphotographs showing changes in Fura-2 emissions resulting from excitation at 340 and 380 nm for the control group. The elevation in the Fura-2 ratio (340/380 ratio) represents an increase in cytosolic Ca2+ concentration. Scale bar 10 µm
10571_2020_996_MOESM2_ESM.tif
(TIF 48504 kb) Supplementary Fig. 2 Representative microphotographs of JC1 fluorescence in a hippocampal slice for the control group. Left: infrared differential interference contrast image; Middle: green fluorescent image excited at 477 nm; Right: red fluorescent image excited at 548 nm. Scale bar 10 µm
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Morisaki, Y., Nakagawa, I., Ogawa, Y. et al. Ischemic Postconditioning Reduces NMDA Receptor Currents Through the Opening of the Mitochondrial Permeability Transition Pore and KATP Channel in Mouse Neurons. Cell Mol Neurobiol 42, 1079–1089 (2022). https://doi.org/10.1007/s10571-020-00996-y
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DOI: https://doi.org/10.1007/s10571-020-00996-y