Two distinct mechanisms are involved in 6-hydroxydopamine- and MPP+- induced dopaminergic neuronal cell death: Role of caspases, ROS, and JNK

Won Seok Choi, So Young Yoon, Tae H. Oh, Eui Ju Choi, Karen L. O'Malley, Young J. Oh

Research output: Contribution to journalArticle

218 Citations (Scopus)

Abstract

In this study, we examined the possibility that MPTP and 6- hydroxydopamine (6-OHDA) act on distinct cell death pathways in a murine dopaminergic neuronal cell line, MN9D. First, we found that cells treated with 6-OHDA accompanied ultrastructural changes typical of apoptosis, whereas MPP+ treatment induced necrotic manifestations. Proteolytic cleavage of poly(ADP-ribose)polymerase by caspase was induced by 6-OHDA, whereas it remained uncleaved up to 32 h after MPP+ treatment and subsequently disappeared. Accordingly, 6-OHDA- but not MPP+-induced cell death was significantly attenuated in the presence of a broad-spectrum caspase inhibitor, N-benzyloxy-carbonyl-Val-Ala-Asp-fluomethylketone (Z-VAD-fmk). As measured by fluorometric probes, the level of reactive oxygen species (ROS) significantly increased after 6-OHDA treatment. In contrast, the level of dihydroethidium-sensitive ROS following MPP+ treatment remained unchanged while a slight increase in dichlorofluorescin-sentive ROS was temporarily observed. As demonstrated by immunoblot analysis, the level of superoxide dismutase was down-regulated following 6-OHDA treatment, whereas it remained unchanged after MPP+ treatment. Cotreatment of cells with antioxidants such as N-acetylcysteine or Mn(III)tetrakis(4-benzoic acid)porphyrin chloride (MnTBAP, cell-permeable superoxide dismutase mimetic) rescued 6-OHDA- but not MPP+-induced cell death, whereas inclusion of catalase or N(G)-nitro-L- arginine had no effect in both cases. In addition, 6-OHDA induced ROS- mediated c-Jun N-terminal kinase (JNK) activation that was attenuated in the presence of N-acetylcysteine or MnTBAP but not catalase or Z-VAD-fmk. In contrast, MPP+ has little effect on JNK activity, indicating that ROS and/or ROS-induced cell death signaling pathway seems to play an essential role in 6-OHDA-mediated apoptosis but not in MPP+-induced necrosis in a mesencephalon-derived, dopaminergic neuronal cell line.

Original languageEnglish
Pages (from-to)86-94
Number of pages9
JournalJournal of Neuroscience Research
Volume57
Issue number1
DOIs
Publication statusPublished - 1999 Jul 1

Keywords

  • Apoptosis
  • Caspase
  • JNK
  • Parkinson's disease
  • Reactive oxygen species

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience

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