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Figure 4 | Journal of Neuroinflammation

Figure 4

From: Cellular injury and neuroinflammation in children with chronic intractable epilepsy

Figure 4

Quantification and Triple immunofluorescence confocal images of DNA fragmentation in the cortex and white matter. (A) DNA fragmentation in individual patients (patients No. 1 to 11) and controls (n = 5). Notice that the majority of our patients [6/10, except RE (No. 11)] showed diffuse cell injury that is comparable to the patient with Rasmussen's encephalitis (No. 11). (B) Fold changes in DNA fragmentation in epilepsy subgroups over controls. (C) The magnitude of DNA fragmentation is significantly correlated with seizure frequency prior to surgery (p = 0.002). (D) Low magnification view of a control shows no evidence of DNA fragmentation. (E-G) Epilepsy case. Low and high magnification of cortex (F) and white matter (G) show increased DNA fragmentations. (H-K) Higher magnification view of DNA fragmentation in neurons (black arrows), oligodendrocytes (green arrows) and astrocytes (blue arrows) in cortex (H, I and J) and white matter (K). Microglia were spared (red arrows). The identification of different cell types was made based on nuclear morphology. Neurons have large vesicular nuclei and oligodendrocytes have round uniform nuclei while astrocytes have bipolar elongated nuclei with irregular borders. Small rod-shaped nuclei of microglia show only faint staining. (L-O) Fluoro-Jade B staining in control (L) and epilepsy cases (M-O). Positive staining in the cortex (M, N) and white matter (O) of epilepsy cases and not in controls are consistent with cell injury noted by in situ end labeling of DNA fragmentation. (P-S) Triple immunofluorescence confocal images of cellular subtypes showing DNA fragmentation in the cortex (P, R) and white matter (Q, S) of focal cortical dysplasia patient; Low magnification (P) shows that neurons comprise the majority of cells with DNA fragmentation. Double labeling of neuron and DNA fragmentation is magnified and microglia closely associated with these neurons are also labeled with fragmented DNA (Fig. R4). DNA fragmentation (blue); microglia (red); neuron (green). (P, R) White matter. DNA fragmentation (blue); astrocyte (red); neuron (green). Low (Q) and high magnification (S) show double labeling of astrocytes with DNA fragmentation. Bars: D-G = 100 μ m; H-S = 20 μ m. * p < 0.05, ** p < 0.01, *** p < 0.001, one-way ANOVA.

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