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 24h with significant increases as early as 12h 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.
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April 2003
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Research Article|
April 01 2003
1-Methyl-4-phenylpyridinium (MPP+)-induced apoptosis and mitochondrial oxidant generation: role of transferrin-receptor-dependent iron and hydrogen peroxide
Shasi V. KALIVENDI;
Shasi V. KALIVENDI
∗Biophysics Research Institute and Free Radical Research Center, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, U.S.A.,
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Srigiridhar KOTAMRAJU;
Srigiridhar KOTAMRAJU
∗Biophysics Research Institute and Free Radical Research Center, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, U.S.A.,
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Sonya CUNNINGHAM;
Sonya CUNNINGHAM
†Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, U.S.A.
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Tiesong SHANG;
Tiesong SHANG
∗Biophysics Research Institute and Free Radical Research Center, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, U.S.A.,
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Cecilia J. HILLARD;
Cecilia J. HILLARD
†Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, U.S.A.
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B. KALYANARAMAN
B. KALYANARAMAN
1
∗Biophysics Research Institute and Free Radical Research Center, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, U.S.A.,
1To whom correspondence should be addressed (e-mail balarama@mcw.edu).
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Publisher: Portland Press Ltd
Received:
October 01 2002
Revision Received:
December 18 2002
Accepted:
January 10 2003
Accepted Manuscript online:
January 10 2003
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2003
2003
Biochem J (2003) 371 (1): 151–164.
Article history
Received:
October 01 2002
Revision Received:
December 18 2002
Accepted:
January 10 2003
Accepted Manuscript online:
January 10 2003
Citation
Shasi V. KALIVENDI, Srigiridhar KOTAMRAJU, Sonya CUNNINGHAM, Tiesong SHANG, Cecilia J. HILLARD, B. KALYANARAMAN; 1-Methyl-4-phenylpyridinium (MPP+)-induced apoptosis and mitochondrial oxidant generation: role of transferrin-receptor-dependent iron and hydrogen peroxide. Biochem J 1 April 2003; 371 (1): 151–164. doi: https://doi.org/10.1042/bj20021525
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