The mitochondrial permeability transition pore and nitric oxide synthase mediate early mitochondrial depolarization in astrocytes during oxygen-glucose deprivation

Recent studies suggest that the degree of mitochondrial dysfunction in cere bral ischemia may be an important determinant of the final extent of tissue injury. Although loss of mitochondrial membrane potential (psi (m)), one i ndex of mitochondrial dysfunction, has been documented in neurons exposed t o ischemic conditions, it is not yet known whether astrocytes, which are re latively resistant to ischemic injury, experience changes in psi (m) under similar conditions. To address this, we exposed cortical astrocytes culture d alone or with neurons to oxygen-glucose deprivation (OGD) and monitored p si (m) using tetramethylrhodamine ethyl ester. Both neurons and astrocytes exhibited profound loss of psi (m) after 45-60 min of OGD. However, althoug h this exposure is lethal to nearly all neurons, it is hours less than that needed to kill astrocytes. Astrocyte psi (m) was rescued during OGD by cyc losporin A, a permeability transition pore blocker, and N-G-nitro-arginine, a nitric oxide synthase inhibitor. Loss of mitochondrial membrane potentia l in astrocytes was not accompanied by depolarization of the plasma membran e. Recovery of astrocyte psi (m) after reintroduction of O-2 and glucose oc curred over a surprisingly long period (>1 hr), suggesting that OGD caused specific, reversible changes in astrocyte mitochondrial physiology beyond t he simple lack of O-2 and glucose. Decreased psi (m) was associated with a cyclosporin A-sensitive loss of cytochrome c but not with activation of cas pase-3 or caspase-9. Our data suggest that astrocyte mitochondrial depolari zation could be a previously unrecognized event early in ischemia and that strategies that target the mitochondrial component of ischemic injury may b enefit astrocytes as well as neurons.