Microarray analysis of oxidative stress regulated genes in mesencephalic dopaminergic neuronal cells: Relevance to oxidative damage in Parkinson's disease

Abstract

Oxidative stress and apoptotic cell death have been implicated in the dopaminergic cell loss that characterizes Parkinson's disease. While factors contributing to apoptotic cell death are not well characterized, oxidative stress is known to activate an array of cell signaling molecules that participate in apoptotic cell death mechanisms. We investigated oxidative stress-induced cytotoxicity of hydrogen peroxide (H2O2) in three cell lines, the dopaminergic mesencephalon-derived N27 cell line, the GABAergic striatum-derived M213-20 cell line, and the hippocampal HN2-5 cell line. N27 cells were more sensitive to H2O2-induced cell death than M213-20 and HN2-5 cells. H2O2 induced significantly greater increases in caspase-3 activity in N27 cells than in M213-20 cells. H2O2-induced apoptotic cell death in N27 cells was mediated by caspase-3-dependent proteolytic activation of PKCdelta. Gene expression microarrays were employed to examine the specific transcriptional changes in N27 cells exposed to 100 microM H2O2 for 4 h. Changes in genes encoding pro- or anti-apoptotic proteins included up-regulation of BIK, PAWR, STAT5B, NPAS2, Jun B, MEK4, CCT7, PPP3CC, and PSDM3, while key down-regulated genes included BNIP3, NPTXR, RAGA, STK6, YWHAH, and MAP2K1. Overall, the changes indicate a modulation of transcriptional activity, chaperone activity, kinase activity, and apoptotic activity that appears highly specific, coordinated and relevant to cell survival. Utilizing this in vitro model to identify novel oxidative stress-regulated genes may be useful in unraveling the molecular mechanisms underlying dopaminergic degeneration in Parkinson's disease.

PAR bZIP-bik is a novel transcriptional pathway that mediates oxidative stress-induced apoptosis in fibroblasts

“…It is worth noting that treatment of neural cell lines with H 2 O 2 upregulated the mRNA levels of bik as determined in microarray experiments, 23 suggesting a proapoptotic role of Bik in oxidative stress-induced neuronal apoptosis. In our fibroblast cell model, bik was constitutively expressed and did not show significant variation in response to H 2 O 2 .…”

Single-Cell Redox Imaging Demonstrates a Distinctive Response of Dopaminergic Neurons to Oxidative Insults

“…Additionally, thiol oxidation has been involved in PD pathogenic events such as K-ATP-mediated neuronal depolarization (3,16,43) and transferring-mediated iron accumulation (45). Therefore, investigating the variations in thiol redox state and the associated signaling events will provide crucial information for understanding the healthy and diseased brain.…”

Exogenous Hsc70, but not thermal preconditioning, confers protection to motoneurons subjected to oxidative stress

“…Hsp25 was upregulated during heat stress, but not during oxidative stress, reflecting what we observed with the other Hsps (data not shown). Additionally, oxidative stress can lead to the activation of glutathione, catalase, superoxide-dismutase, ubiquitin-proteosome components, and many other proteins involved in apoptosis, cell cycle, intracellular signaling, and transcription (Halliwell and Gutteridge, 1999;Anantharam et al, 2007). Despite the possible upregulation of other proteins due to oxi- Figure 5 Characterization of rhHsc70 and its domains.…”