1-Methyl-4-phenylpyridinium-induced Apoptosis in Cerebellar Granule Neurons Is Mediated by Transferrin Receptor Iron-dependent Depletion of Tetrahydrobiopterin and Neuronal Nitric-oxide Synthase-derived Superoxide

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

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Abstract

In this study, we investigated the molecular mechanisms of toxicity of 1-methyl-4-phenylpyridinium (MPP+), an ultimate toxic metabolite of a mitochondrial neurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, that causes Parkinson-like symptoms in experimental animals and humans. We used rat cerebellar granule neurons as a model cell system for investigating MPP + toxicity. Results show that MPP+ treatment resulted in the generation of reactive oxygen species from inhibition of complex I of the mitochondrial respiratory chain, and inactivation of aconitase. This, in turn, stimulated transferrin receptor (TfR)-dependent iron signaling via activation of the iron-regulatory protein/iron-responsive element interaction. MPP + caused a time-dependent depletion of tetrahydrobiopterin (BH 4) that was mediated by H2O2 and transferrin iron. Depletion of BH4 decreased the active, dimeric form of neuronal nitric-oxide synthase (nNOS). MPP+-mediated "uncoupling" of nNOS decreased .NO and increased superoxide formation. Pretreatment of cells with sepiapterin to promote BH 4 biosynthesis or cell-permeable iron chelator and TfR antibody to prevent iron-catalyzed BH4 decomposition inhibited MPP+ cytotoxicity. Preincubation of cerebellar granule neurons with nNOS inhibitor exacerbated MPP+-induced iron uptake, BH4 depletion, proteasomal inactivation, and apoptosis. We conclude that MPP +-dependent aconitase inactivation, Tf-iron uptake, and oxidant generation result in the depletion of intracellular BH4, leading to the uncoupling of nNOS activity. This further exacerbates reactive oxygen species-mediated oxidative damage and apoptosis. Implications of these results in unraveling the molecular mechanisms of neurodegenerative diseases (Parkinson's and Alzheimer's disease) are discussed.

Original languageEnglish (US)
Pages (from-to)19099-19112
Number of pages14
JournalJournal of Biological Chemistry
Volume279
Issue number18
DOIs
StatePublished - Apr 30 2004

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1-Methyl-4-phenylpyridinium
Nitric Oxide Synthase Type I
Transferrin Receptors
Superoxides
Neurons
Iron
Apoptosis
Aconitate Hydratase
Toxicity
Reactive Oxygen Species
Iron-Regulatory Proteins
Neurodegenerative diseases
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine
Poisons
Biosynthesis
Neurotoxins
Cytotoxicity
Transferrin
Chelating Agents
Electron Transport

ASJC Scopus subject areas

  • Biochemistry

Cite this

1-Methyl-4-phenylpyridinium-induced Apoptosis in Cerebellar Granule Neurons Is Mediated by Transferrin Receptor Iron-dependent Depletion of Tetrahydrobiopterin and Neuronal Nitric-oxide Synthase-derived Superoxide. / Shang, Tiesong; Kotamraju, Srigiridhar; Kalivendi, Shasi V.; Hillard, Cecilia J.; Kalyanaraman, B.

In: Journal of Biological Chemistry, Vol. 279, No. 18, 30.04.2004, p. 19099-19112.

Research output: Contribution to journalArticle

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abstract = "In this study, we investigated the molecular mechanisms of toxicity of 1-methyl-4-phenylpyridinium (MPP+), an ultimate toxic metabolite of a mitochondrial neurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, that causes Parkinson-like symptoms in experimental animals and humans. We used rat cerebellar granule neurons as a model cell system for investigating MPP + toxicity. Results show that MPP+ treatment resulted in the generation of reactive oxygen species from inhibition of complex I of the mitochondrial respiratory chain, and inactivation of aconitase. This, in turn, stimulated transferrin receptor (TfR)-dependent iron signaling via activation of the iron-regulatory protein/iron-responsive element interaction. MPP + caused a time-dependent depletion of tetrahydrobiopterin (BH 4) that was mediated by H2O2 and transferrin iron. Depletion of BH4 decreased the active, dimeric form of neuronal nitric-oxide synthase (nNOS). MPP+-mediated {"}uncoupling{"} of nNOS decreased .NO and increased superoxide formation. Pretreatment of cells with sepiapterin to promote BH 4 biosynthesis or cell-permeable iron chelator and TfR antibody to prevent iron-catalyzed BH4 decomposition inhibited MPP+ cytotoxicity. Preincubation of cerebellar granule neurons with nNOS inhibitor exacerbated MPP+-induced iron uptake, BH4 depletion, proteasomal inactivation, and apoptosis. We conclude that MPP +-dependent aconitase inactivation, Tf-iron uptake, and oxidant generation result in the depletion of intracellular BH4, leading to the uncoupling of nNOS activity. This further exacerbates reactive oxygen species-mediated oxidative damage and apoptosis. Implications of these results in unraveling the molecular mechanisms of neurodegenerative diseases (Parkinson's and Alzheimer's disease) are discussed.",
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T1 - 1-Methyl-4-phenylpyridinium-induced Apoptosis in Cerebellar Granule Neurons Is Mediated by Transferrin Receptor Iron-dependent Depletion of Tetrahydrobiopterin and Neuronal Nitric-oxide Synthase-derived Superoxide

AU - Shang, Tiesong

AU - Kotamraju, Srigiridhar

AU - Kalivendi, Shasi V.

AU - Hillard, Cecilia J.

AU - Kalyanaraman, B.

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AB - In this study, we investigated the molecular mechanisms of toxicity of 1-methyl-4-phenylpyridinium (MPP+), an ultimate toxic metabolite of a mitochondrial neurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, that causes Parkinson-like symptoms in experimental animals and humans. We used rat cerebellar granule neurons as a model cell system for investigating MPP + toxicity. Results show that MPP+ treatment resulted in the generation of reactive oxygen species from inhibition of complex I of the mitochondrial respiratory chain, and inactivation of aconitase. This, in turn, stimulated transferrin receptor (TfR)-dependent iron signaling via activation of the iron-regulatory protein/iron-responsive element interaction. MPP + caused a time-dependent depletion of tetrahydrobiopterin (BH 4) that was mediated by H2O2 and transferrin iron. Depletion of BH4 decreased the active, dimeric form of neuronal nitric-oxide synthase (nNOS). MPP+-mediated "uncoupling" of nNOS decreased .NO and increased superoxide formation. Pretreatment of cells with sepiapterin to promote BH 4 biosynthesis or cell-permeable iron chelator and TfR antibody to prevent iron-catalyzed BH4 decomposition inhibited MPP+ cytotoxicity. Preincubation of cerebellar granule neurons with nNOS inhibitor exacerbated MPP+-induced iron uptake, BH4 depletion, proteasomal inactivation, and apoptosis. We conclude that MPP +-dependent aconitase inactivation, Tf-iron uptake, and oxidant generation result in the depletion of intracellular BH4, leading to the uncoupling of nNOS activity. This further exacerbates reactive oxygen species-mediated oxidative damage and apoptosis. Implications of these results in unraveling the molecular mechanisms of neurodegenerative diseases (Parkinson's and Alzheimer's disease) are discussed.

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