BNIP3 up-regulation and mitochondrial dysfunction in manganese-induced neurotoxicity.
The central nervous system (CNS) appears to be the critical target of manganese (Mn), and neurotoxicity has been the focus of most of the health effects of manganese. In brain, the mechanism underlying the Mn-induced cell death is not clear. We have previously demonstrated that NFkappabeta induction and the activation of nitric oxide synthase through reactive oxygen species (ROS) represent a proximate mechanism for Mn-induced neurotoxicity. In this study, an immortalized dopaminergic cells were used to characterize the cell death signaling cascade activated by manganese. Exposure to Mn resulted in a time and concentration-related loss of cell viability as observed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and live/dead cell assay. Mn increased BNIP3 expression within 3h and continued to increase up to 24h exposure followed by a concentration-related apoptotic death as determined by TUNEL. Further, Mn treatment resulted in accumulation of reactive oxygen species and mitochondrial dysfunction with loss of mitochondrial membrane potential and release of cytochrome c. Antioxidants significantly reduced Mn-induced BNIP3 expression and attenuated cell death, demonstrating the role of oxidative stress in BNIP3 induction. Blocking BNIP3 up-regulation with a transcription or a translational inhibitor reduced the response to manganese. Cell death by manganese was reduced in the presence of CsA (PT pore inhibitor). In addition, knockdown of BNIP3 by small interfering RNA (siRNA) improved mitochondrial recovery and reduced neuronal cell loss suggesting that constitutive expression of BNIP3 plays a role in Mn-induced neurotoxicity by regulating mitochondrial functions. These findings indicate a potential detrimental role of BNIP3 in manganese-induced neuronal cell death.
Prabhakaran
Krishnan
K
Naval Health Research Center Detachment, Environmental Health Effects Laboratory, Wright-Patterson Air Force Base, 2729 R Street, Area B, Building 837, Dayton, OH 45433, USA.
Chapman
Gail D
GD
Gunasekar
Palur G
PG
D000818:Animals; D017209:Apoptosis; D045744:Cell Line, Tumor; D016572:Cyclosporine / Q000494:pharmacology; D045304:Cytochromes c / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D008345:Manganese / Q000037:antagonists & inhibitors / Q000633:toxicity*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects*; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D051336:Mitochondrial Membranes / Q000187:drug effects* / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000378:metabolism*; D009474:Neurons / Q000187:drug effects* / Q000648:ultrastructure; D018384:Oxidative Stress / Q000187:drug effects; D051381:Rats; D014131:Trace Elements / Q000037:antagonists & inhibitors / Q000633:toxicity*; D015854:Up-Regulation
C508686:BNip3 protein, mouse; D004791:Enzyme Inhibitors; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D014131:Trace Elements; D008345:Manganese; D016572:Cyclosporine; D045304:Cytochromes c