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

Research output: Contribution to journalArticle

51 Scopus citations


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
Issue number18
StatePublished - Apr 30 2004


ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this