1-Methyl-4-phenylpyridinium (MPP+)-induced apoptosis and mitochondrial oxidant generation: role of transferrin-receptor-dependent iron and hydrogen peroxide.

Abstract

1-Methyl-4-phenylpyridinium (MPP(+)) is a neurotoxin used in cellular models of Parkinson's Disease. Although intracellular iron plays a crucial role in MPP(+)-induced apoptosis, the molecular signalling mechanisms linking iron, reactive oxygen species (ROS) and apoptosis are still unknown. We investigated these aspects using cerebellar granule neurons (CGNs) and human SH-SY5Y neuroblastoma cells. MPP(+) enhanced caspase 3 activity after 24 h with significant increases as early as 12 h after treatment of cells. Pre-treatment of CGNs and neuroblastoma cells with the metalloporphyrin antioxidant enzyme mimic, Fe(III)tetrakis(4-benzoic acid)porphyrin (FeTBAP), completely prevented the MPP(+)-induced caspase 3 activity as did overexpression of glutathione peroxidase (GPx1) and pre-treatment with a lipophilic, cell-permeable iron chelator [N, N '-bis-(2-hydroxybenzyl)ethylenediamine-N, N '-diacetic acid, HBED]. MPP(+) treatment increased the number of TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labelling)-positive cells which was completely blocked by pre-treatment with FeTBAP. MPP(+) treatment significantly decreased the aconitase and mitochondrial complex I activities; pre-treatment with FeTBAP, HBED and GPx1 overexpression reversed this effect. MPP(+) treatment increased the intracellular oxidative stress by 2-3-fold, as determined by oxidation of dichlorodihydrofluorescein and dihydroethidium (hydroethidine). These effects were reversed by pre-treatment of cells with FeTBAP and HBED and by GPx1 overexpression. MPP(+)-treatment enhanced the cell-surface transferrin receptor (TfR) expression, suggesting a role for TfR-induced iron uptake in MPP(+) toxicity. Treatment of cells with anti-TfR antibody (IgA class) inhibited MPP(+)-induced caspase activation. Inhibition of nitric oxide synthase activity did not affect caspase 3 activity, apoptotic cell death or ROS generation by MPP(+). Overall, these results suggest that MPP(+)-induced cell death in CGNs and neuroblastoma cells proceeds via apoptosis and involves mitochondrial release of ROS and TfR-dependent iron.

• Authors: Shasi V Kalivendi, Srigiridhar Kotamraju, Sonya Cunningham, Tiesong Shang, Cecilia J Hillard, B Kalyanaraman
• Journal: The Biochemical journal (Biochem J) Vol. 371 Issue Pt 1 Pg. 151-64 (Apr 01 2003) ISSN: 0264-6021 [Print] England
• PMID: 12523938 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Antioxidants
• Iron Chelating Agents
• Metalloporphyrins
• Oxidants
• Reactive Oxygen Species
• Receptors, Transferrin
• Transferrin
• tetrakis(4-benzoic acid)porphyriniron(III)
• N,N'-bis(2-hydroxybenzyl)ethylenediamine-N,N'-diacetic acid
• Edetic Acid
• Hydrogen Peroxide
• Iron
• Glutathione Peroxidase
• NADH, NADPH Oxidoreductases
• CASP3 protein, human
• Caspase 3
• Caspases
• Aconitate Hydratase
• Deferoxamine
• 1-Methyl-4-phenylpyridinium
• Glutathione Peroxidase GPX1
• GPX1 protein, human

Topics

• 1-Methyl-4-phenylpyridinium (pharmacology)
• Aconitate Hydratase (drug effects, metabolism)
• Animals
• Antioxidants (pharmacology)
• Apoptosis (drug effects)
• Caspase 3
• Caspases (drug effects, metabolism)
• Cells, Cultured
• Deferoxamine (pharmacology)
• Edetic Acid (analogs & derivatives, pharmacology)
• Female
• Glutathione Peroxidase (drug effects, genetics, metabolism)
• Humans
• Hydrogen Peroxide (metabolism)
• Iron (metabolism)
• Iron Chelating Agents (pharmacology)
• Male
• Metalloporphyrins (pharmacology)
• Mitochondria (drug effects, metabolism)
• NADH, NADPH Oxidoreductases (metabolism)
• Neurons (drug effects, metabolism)
• Oxidants (metabolism)
• Oxidative Stress
• Parkinson Disease (metabolism, pathology)
• Reactive Oxygen Species (metabolism)
• Receptors, Transferrin (drug effects, immunology, metabolism)
• Staining and Labeling (methods)
• Transferrin (metabolism)
• Glutathione Peroxidase GPX1