Glucocorticoid-induced apoptosis of thymocytes: requirement of proteasome-dependent mitochondrial activity.
Thymocytes undergo negative and positive selection during development in the thymus. During this selection process, the majority of thymocytes are eliminated by apoptosis through signaling via TCR or die by neglect, possibly mediated through glucocorticoids. In this study, we report that thymocytes require molecular oxygen to undergo apoptosis induced by dexamethasone (DEX), a synthetic glucocorticoid, and treatment with N-acetyl-L-cysteine (NAC), a thiol antioxidant, inhibits thymocyte apoptosis in vivo as well as ex vivo. We detected elevated intracellular levels of hydrogen peroxide (H(2)O(2)) during DEX-induced apoptosis, which is reduced by NAC treatment, indicating that the elevated levels of intracellular H(2)O(2) are proapoptotic. We also show that loss of mitochondrial membrane potential, cytochrome c release, as well as caspase-3 activation induced by DEX are attenuated by NAC treatment. We identified the production site for H(2)O(2) as the ubiquinone cycle at complex III of mitochondria by using various inhibitors of the mitochondrial electron transport chain, and we show that the cell death events mediated by mitochondria are also significantly reduced when the inhibitors were used. Through inhibition of the proteasome, we also show that the production of H(2)O(2) and the cell death events mediated by mitochondria are regulated by proteosomal activities in DEX-induced thymocyte apoptosis. We conclude that in DEX-treated thymocytes, the increased production of H(2)O(2) originates from mitochondria and is proapoptotic for cell death mediated by mitochondria. We also conclude that all the apoptotic events mediated by mitochondria are regulated by proteasomes.
Tonomura
Noriko
N
Department of Veterinary and Animal Sciences and Program in Molecular and Cellular Biology, University of Massachusetts, Amherst, MA 01003, USA.
McLaughlin
Kelly
K
Grimm
Lisa
L
Goldsby
Richard A
RA
Osborne
Barbara A
BA
D000111:Acetylcysteine / Q000008:administration & dosage*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000276:immunology; D016923:Cell Death / Q000276:immunology; D002478:Cells, Cultured; D003546:Cysteine Endopeptidases / Q000378:metabolism / Q000502:physiology*; D003907:Dexamethasone / Q000008:administration & dosage*; D005260:Female; D006861:Hydrogen Peroxide / Q000378:metabolism; D007274:Injections, Intraperitoneal; D051379:Mice; D008807:Mice, Inbred BALB C; D008928:Mitochondria / Q000187:drug effects / Q000201:enzymology* / Q000276:immunology / Q000378:metabolism; D009097:Multienzyme Complexes / Q000378:metabolism / Q000502:physiology*; D010100:Oxygen / Q000502:physiology; D046988:Proteasome Endopeptidase Complex; D017382:Reactive Oxygen Species / Q000494:pharmacology; D013950:Thymus Gland / Q000166:cytology* / Q000187:drug effects* / Q000201:enzymology / Q000276:immunology
D009097:Multienzyme Complexes; D017382:Reactive Oxygen Species; D003907:Dexamethasone; D006861:Hydrogen Peroxide; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; D010100:Oxygen; D000111:Acetylcysteine