Bax affects production of reactive oxygen by the mitochondria of non-apoptotic neurons.
Depriving sympathetic neurons in cell culture of nerve growth factor (NGF) causes their apoptotic death. Bax-induced release of cytochrome c from mitochondria and the subsequent activation of cytosolic caspases are central to this death. A Bax-dependent increase of mitochondrial-derived reactive oxygen species (ROS) that is an important component of the apoptotic cascade in these cells begins soon after NGF withdrawal. Here we report that Bax can also influence mitochondrial production of ROS in non-apoptotic sympathetic neurons. We determined ROS levels by using confocal microscopy to monitor changes in the fluorescence intensity of a redox-sensitive dye loaded into single cells. ROS levels were similar in NGF-replete bax wild-type neurons and neurons from which bax had been deleted. To enhance any effects that Bax might have on ROS levels in NGF-replete cells we exposed cultures to the ATP synthase inhibitor, oligomycin. This treatment hyperpolarizes mitochondrial membrane potential (DeltaPsi(m)), an event that can favor increased ROS production. NGF-replete neurons from mice in which bax had been deleted had much higher levels of mitochondrial-derived ROS when treated with oligomycin than did bax wild-type cells. Oligomycin treatment also caused greater hyperpolarization of DeltaPsi(m) in bax-deleted cells than in wild-type cells. These findings indicate that Bax can affect mitochondrial ROS production in non-apoptotic neurons and may do so by altering DeltaPsi(m).
Kirkland
Rebecca A
RA
Department of Pharmaceutical and Biomedical Sciences, University of Georgia College of Pharmacy, 357 Wilson Pharmacy, Athens, GA 30602, USA.
Franklin
James L
JL
D000818:Animals; D017209:Apoptosis; D004791:Enzyme Inhibitors / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D018345:Mice, Knockout; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000378:metabolism* / Q000502:physiology; D051336:Mitochondrial Membranes / Q000378:metabolism* / Q000502:physiology; D025261:Mitochondrial Proton-Translocating ATPases / Q000037:antagonists & inhibitors; D009414:Nerve Growth Factors / Q000378:metabolism; D009474:Neurons / Q000378:metabolism* / Q000502:physiology; D009840:Oligomycins / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D017783:Superior Cervical Ganglion / Q000166:cytology; D051028:bcl-2-Associated X Protein / Q000172:deficiency / Q000378:metabolism*
D004791:Enzyme Inhibitors; D009414:Nerve Growth Factors; D009840:Oligomycins; D017382:Reactive Oxygen Species; D051028:bcl-2-Associated X Protein; D025261:Mitochondrial Proton-Translocating ATPases