title: Anoxia-mediated calcium release through the mitochondrial permeability transition pore silences NMDA receptor currents in turtle neurons.
abstract: Mammalian neurons are anoxia sensitive and rapidly undergo excitotoxic cell death when deprived of oxygen, mediated largely by Ca(2+) entry through over-activation of N-methyl-d-aspartate receptors (NMDARs). This does not occur in neurons of the anoxia-tolerant western painted turtle, where a decrease in NMDAR currents is observed with anoxia. This decrease is dependent on a modest rise in cytosolic [Ca(2+)] ([Ca(2+)]c) that is mediated by release from the mitochondria. The aim of this study was to determine whether the mitochondrial permeability transition pore (mPTP) is involved in NMDAR silencing through release of mitochondrial Ca(2+). Opening the mPTP during normoxia with atractyloside decreased NMDAR currents by releasing mitochondrial Ca(2+), indicated by an increase in Oregon Green fluorescence. Conversely, the mPTP blocker cyclosporin A prevented the anoxia-mediated increase in [Ca(2+)]c and reduction in NMDAR currents. Mitochondrial membrane potential (Ψm) was determined using rhodamine-123 fluorescence and decreased with the onset of anoxia in a time frame that coincided with the increase in [Ca(2+)]c. Activation of mitochondrial ATP-sensitive potassium (mK(+)ATP) channels also releases mitochondrial Ca(2+) and we show that activation of mK(+)ATP channels during normoxia with diazoxide leads to Ψm depolarization and inhibition with 5-hydroxydecanoic acid blocked anoxia-mediated Ψm depolarization. Ψm does not collapse during anoxia but rather reaches a new steady-state level that is maintained via ATP hydrolysis by the F1-F0 ATPase, as inhibition with oligomycin depolarizes Ψm further than the anoxic level. We conclude that anoxia activates mK(+)ATP channels, which leads to matrix depolarization, Ca(2+) release via the mPTP, and ultimately silencing of NMDARs.
authors.affiliation: Department of Cell and Systems Biology, University of Toronto, Toronto, ON, Canada, M5S 3G5.
authors.forename: Peter John
authors.initials: PJ
authors.lastname: Hawrysh
authors.forename: Leslie Thomas
authors.initials: LT
authors.lastname: Buck
mesh_terms: D000818:Animals; D020013:Calcium Signaling; D015687:Cell Hypoxia; D054086:KATP Channels / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial; D033681:Mitochondrial Membrane Transport Proteins / Q000378:metabolism / Q000502:physiology*; D000083162:Mitochondrial Permeability Transition Pore; D008954:Models, Biological; D009474:Neurons / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism / Q000502:physiology*; D014426:Turtles / Q000378:metabolism*
chemical_list: D054086:KATP Channels; D033681:Mitochondrial Membrane Transport Proteins; D000083162:Mitochondrial Permeability Transition Pore; D016194:Receptors, N-Methyl-D-Aspartate
keywords: anoxia tolerance; channel arrest; mitochondrial BK channel; mitochondrial membrane potential; pyramidal neurons; whole-cell patch-clamp