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Mode of cell death in C6 glial cell exposed to 1‐methyl‐4‐phenylpyridinum (MPP+)
Author(s) -
Badisa Ramesh B,
Williams Zakia R,
DarlingReed Selina,
Soliman Karam F
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.707.19
Subject(s) - propidium iodide , astrogliosis , programmed cell death , apoptosis , necrosis , annexin , flow cytometry , chemistry , cell , microbiology and biotechnology , biology , pathology , medicine , neuroscience , biochemistry , central nervous system
1‐methyl‐4‐phenylpyridinum (MPP + ) is shown to cause selective death of nigrostriatal dopaminergic neurons in the brain, and is widely employed to induce Parkinson's disease symptoms in animal models. The present study was designed to examine if MPP + ‐induced astrogliosis is related to apoptosis or necrosis in presence of 2 or 10 mM D‐glucose by using flowcytometry. For this purpose, the C6 glial cell line was used as an in vitro model to assess the MPP + ‐induced astrogliosis by testing at 10, 30, 50, 70, 90, 100, 150, 250, 500 and 1000 μM for 12 h. The treated cells were stained with annexin V and propidium iodide (PI) and analyzed by flowcytometry within 1 h to assess the mode of cell death. In order to investigate the effect of MPP + on cell cycle stages, the rat C6 glial cells were treated with MPP + at 100, 150, 250, 500 and 1000 μM for 12 h in the presence of 2 or 10 mM D‐glucose. These cells were stained with PI and analyzed by flow‐cytometry. The data indicate that exposure with MPP + at 100 μM or above caused necrotic cell death predominantly when cells were grown in 2 mM D‐glucose, while MPP + atlower concentrations of 10, 30, 50, 70 and 90 μM caused significantly higher apoptotic cell death than necrotic. Interestingly, D‐glucose at 10 mM not only decreased the number of cells undergoing necrosis caused by MPP + at 100 μM or higher but also increased the number of live cells at these concentrations. Cell cycle analysis indicate that glial cells in 2 mM D‐glucose were marginally (< 12%) arrested at G1 stage with 1000 μM MPP + , while 10 mM D‐glucose decreased the percentage of cell arrest (< 6%) with 1000 μM MPP + . Moreover, the cell cycle results confirm the protective role of D‐glucose against MPP + toxicity. (Supported by NIH Grant RR03020)