Mitochondrial depolarization in glutamate-stimulated neurons: an early signal specific to excitotoxin exposure.
A brief exposure to high concentrations of glutamate kills cultured forebrain neurons by an excitotoxic process that is dependent on Ca2+ influx through the NMDA receptor. In this study, we have measured striking changes in mitochondrial function during and immediately after intense glutamate receptor activation. Using indo-1 microfluorometry and a specific inhibitor of the mitochondrial Na+/Ca2+ exchanger, CGP-37157, we have demonstrated that mitochondria accumulate large quantities of Ca2+ during a toxic glutamate stimulus and further that Ca2+ efflux from mitochondria contributes to the prolonged [Ca2+]i elevation after glutamate removal. We then used JC-1 (5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine+ ++ iodide), a ratiometric indicator of mitochondrial membrane potential (delta psi), to show that Ca2+ accumulation within the organelle dissipates delta psi. The abrupt loss of delta psi after glutamate stimulation did not occur in the presence of MK801 or in the absence of extracellular Ca2+. The mitochondrial depolarization was also cyclosporin A-sensitive, indicating a probable role for the permeability transition pore. Hence mitochondrial Ca2+ accumulation and the subsequent permeability transition may be a critical early event specific to the NMDA receptor-mediated excitotoxic cascade.
White
R J
RJ
Center for Neuroscience, University of Pittsburgh School of Medicine, Pennsylvania 15261, USA.
Reynolds
I J
IJ
D000818:Animals; D001562:Benzimidazoles / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002232:Carbocyanines / Q000494:pharmacology; D002352:Carrier Proteins / Q000037:antagonists & inhibitors; D002478:Cells, Cultured; D002998:Clonazepam / Q000031:analogs & derivatives / Q000494:pharmacology; D004594:Electrophysiology; D005456:Fluorescent Dyes; D005470:Fluorometry; D018698:Glutamic Acid / Q000494:pharmacology*; D007211:Indoles; D008564:Membrane Potentials / Q000187:drug effects; D008928:Mitochondria / Q000502:physiology*; D009474:Neurons / Q000187:drug effects* / Q000502:physiology*; D009498:Neurotoxins / Q000494:pharmacology*; D016548:Prosencephalon / Q000166:cytology; D051381:Rats; D017470:Receptors, Glutamate / Q000502:physiology; D019831:Sodium-Calcium Exchanger; D013841:Thiazepines / Q000494:pharmacology
D001562:Benzimidazoles; D002232:Carbocyanines; D002352:Carrier Proteins; D005456:Fluorescent Dyes; D007211:Indoles; D009498:Neurotoxins; D017470:Receptors, Glutamate; D019831:Sodium-Calcium Exchanger; D013841:Thiazepines; C068624:5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine; D018698:Glutamic Acid; D002998:Clonazepam; C079032:CGP 37157; C048960:indo-1; D002118:Calcium