TY - JOUR
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.
PY - 2004/4/30
Y1 - 2004/4/30
N2 - 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.
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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U2 - 10.1074/jbc.M400101200
DO - 10.1074/jbc.M400101200
M3 - Article
C2 - 14752097
AN - SCOPUS:2442504779
SN - 0021-9258
VL - 279
SP - 19099
EP - 19112
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 18
ER -