Bax regulates production of superoxide in both apoptotic and nonapoptotic neurons: role of caspases.
A Bax- and, apparently, mitochondria-dependent increase in superoxide (O(2)(·-)) and other reactive oxygen species (ROS) occurs in apoptotic superior cervical ganglion (SCG) and cerebellar granule (CG) neurons. Here we show that Bax also lies upstream of ROS produced in nonapoptotic neurons and present evidence that caspases partially mediate the pro-oxidant effect of Bax. We used the O(2)(·-)-sensitive dye MitoSOX to monitor O(2)(·-) in neurons expressing different levels of Bax and mitochondrial superoxide dismutase (SOD2). Basal and apoptotic O(2)(·-) levels in both SCG and CG neurons were reduced in SOD2 wild-type (WT) cells having lower Bax concentrations. Apoptotic and nonapoptotic neurons from Bax-WT/SOD2-null but not Bax-null/SOD2-null mice had increased O(2)(·-) levels. A caspase inhibitor inhibited O(2)(·-) in both apoptotic and nonapoptotic SCG neurons. O(2)(·-) production increased when WT, but not Bax-null, SCG neurons were permeabilized and treated with active caspase 3. There was no apoptosis and little increase in O(2)(·-) in SCG neurons from caspase 3-null mice exposed to an apoptotic stimulus. O(2)(·-) levels in nonapoptotic caspase 3-null SCG neurons were lower than in WT cells but not as low as in caspase inhibitor-treated cells. These data indicate that Bax lies upstream of most O(2)(·-) produced in neurons, that caspase 3 is required for increased O(2)(·-) production during neuronal apoptosis, that caspase 3 is partially involved in O(2)(·-) production in nonapoptotic neurons, and that other caspases may also be involved in Bax-dependent O(2)(·-) production in nonapoptotic cells.
Department of Pharmaceutical and Biomedical Sciences, The University of Georgia College of Pharmacy, Athens, Georgia 30602, USA.
Rebecca A
RA
Kirkland
Geraldine M
GM
Saavedra
Brian S
BS
Cummings
James L
JL
Franklin
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D053148:Caspase 3 / Q000502:physiology*; D002478:Cells, Cultured; D005260:Female; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D024101:Mitochondrial Proteins / Q000502:physiology; D009474:Neurons / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism*; D051028:bcl-2-Associated X Protein / Q000502:physiology*
C494827:Bax protein, mouse; D024101:Mitochondrial Proteins; D017382:Reactive Oxygen Species; D051028:bcl-2-Associated X Protein; D013481:Superoxides; D053148:Caspase 3