Protection against MPP+ neurotoxicity in cerebellar granule cells by antioxidants

Authors

  • Rosa A. González-Polo,

    1. Departamento de Bioquımica y Biologıa Molecular y Genética, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, Spain
    Search for more papers by this author
    • R.A.G.-P. and G.S. contributed equally to this work.

  • Germán Soler,

    1. Departamento de Bioquımica y Biologıa Molecular y Genética, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, Spain
    Search for more papers by this author
    • R.A.G.-P. and G.S. contributed equally to this work.

  • Andrea Rodrıguezmartın,

    1. Departamento de Bioquımica y Biologıa Molecular y Genética, E.U. Enfermerıa y T.O., Universidad de Extremadura, Avda. de la Universidad s/n 10071, Cáceres, Spain
    Search for more papers by this author
  • Jose M. Morán,

    1. Departamento de Bioquımica y Biologıa Molecular y Genética, E.U. Enfermerıa y T.O., Universidad de Extremadura, Avda. de la Universidad s/n 10071, Cáceres, Spain
    Search for more papers by this author
  • José M. Fuentes

    Corresponding author
    1. Departamento de Bioquımica y Biologıa Molecular y Genética, E.U. Enfermerıa y T.O., Universidad de Extremadura, Avda. de la Universidad s/n 10071, Cáceres, Spain
    Search for more papers by this author

Corresponding author. Tel.: +34-927-257450; fax: +34-927-257451. jfuentes@unex.es

Abstract

The neuropathology associated with Parkinson's disease (PD) is thought to involve excessive production of free radicals, dopamine autoxidation, defects in glutathione peroxidase expression, attenuated levels of reduced glutathione, altered calcium homeostasis, excitotoxicity and genetic defects in mitochondrial complex I activity. While the neurotoxic mechanisms are vastly different for excitotoxins and 1-methyl-4-phenylpyridinium ion (MPP+), both are thought to involve free radical production, compromised mitochondrial activity and excessive lipid peroxidation. We show here that the levels of reactive oxygen species (ROS) and reactive nitrogen species (RNS) increased significantly after treatment of cultured cerebellar granule cells (CGCs) with 50 μM MPP+. Co-treatment with antioxidants such as ascorbate (ASC), catalase, α-tocopherol (α-TOH), coenzyme Q10 (CoQ10) or superoxide dismutase (SOD) rescued the cells from MPP+-induced death. MPP+-induced cell death was also abolished by co-treatment with nitric oxide synthase (NOS) inhibitors such as 7-nitroindazole (7-NI), 2-ethyl-2-thiopseudourea hydrobromide (EPTU) or S-methylisothiourea sulphate (MPTU). We also tested the protective effects of an iron chelator (deferoxamine mesylate, DFx) and a peroxynitrite scavenger (FeTTPS) and the results lend further support to the view that the free radical cytotoxicity plays an essential role in MPP+-induced death in primary cultures of CGC.

Ancillary