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

Shasi V. Kalivendi, Srigiridhar Kotamraju, Sonya Cunningham, Tiesong Shang, Cecilia J. Hillard, B. Kalyanaraman

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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.

Original languageEnglish (US)
Pages (from-to)151-164
Number of pages14
JournalBiochemical Journal
Volume371
Issue number1
DOIs
StatePublished - Apr 1 2003

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1-Methyl-4-phenylpyridinium
Transferrin Receptors
Oxidants
Hydrogen Peroxide
Iron
Apoptosis
Cells
Caspase 3
Neurons
Reactive Oxygen Species
ethylenediamine
Cell death
Neuroblastoma
Metalloporphyrins
Aconitate Hydratase
Oxidative stress
DNA Nucleotidylexotransferase
Neurotoxins
Chelating Agents
Glutathione Peroxidase

Keywords

  • Caspase 3
  • Glutathione peroxidase
  • Metalloporphyrin
  • Nitric oxide synthase
  • Oxidative stress
  • Parkinson's disease

ASJC Scopus subject areas

  • Biochemistry

Cite this

1-Methyl-4-phenylpyridinium (MPP+)-induced apoptosis and mitochondrial oxidant generation : Role of transferrin-receptor-dependent iron and hydrogen peroxide. / Kalivendi, Shasi V.; Kotamraju, Srigiridhar; Cunningham, Sonya; Shang, Tiesong; Hillard, Cecilia J.; Kalyanaraman, B.

In: Biochemical Journal, Vol. 371, No. 1, 01.04.2003, p. 151-164.

Research output: Contribution to journalArticle

Kalivendi, Shasi V. ; Kotamraju, Srigiridhar ; Cunningham, Sonya ; Shang, Tiesong ; Hillard, Cecilia J. ; Kalyanaraman, B. / 1-Methyl-4-phenylpyridinium (MPP+)-induced apoptosis and mitochondrial oxidant generation : Role of transferrin-receptor-dependent iron and hydrogen peroxide. In: Biochemical Journal. 2003 ; Vol. 371, No. 1. pp. 151-164.
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