Key Proteins of Activating Cell Death Can Be Predicted through a Kainic Acid-Induced Excitotoxic Stress

Figure 2

Comparison of mitochondrial membrane potential (MMP) measured by flow cytometry following exposure to kainic acid (KA) over different time periods. (a–e) Representative fluorescence histograms obtained from dissociated cortical neurons (10⁶ cells/mL) from 14-day-old rat brains incubated with 3-3′-dihexyloxacarbocyanine in the presence of 0.005 pM, 0.5 pM, 50 nM, 5 μM, and 500 μM KA for 5 (red), 30 (yellow), 60 (blue), and 180 min (purple). The white curves represent the background fluorescence of untreated neurons; the colored curves represent the increased loss of MMP from cells (indicated by an increase in 535 nm fluorescence intensity versus white curve) following KA treatment. (f) Bar graphs of MMP intensity (mean ± SE) expressed as the percentage change in mean fluorescence intensity (MFI) from controls. Experiments were performed in triplicate, and statistical analyses of the data were performed using ANOVA. Data are expressed as percentage (%) change from controls. compared with the respective time period in the control group. compared with the respective time period in the 0.5 pM KA group. Apoptosis and necrosis labels indicate the degrees of cell death at 180 min of treatment.

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