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. 2010 Dec;177(6):3051–3060. doi: 10.2353/ajpath.2010.100442

Figure 3.

Figure 3

Direct oligodendroglial and collateral neuronal apoptosis in slices from ODC-OVA mice incubated with OVA-specific OT-I effector T cells is reduced by pre-incubation of OT-I splenocytes but not slices with GA. A and C: GA does not prevent neuronal cell death in acute brain slices from ODC-OVA mice on incubation for 6 hours with OT-I effector T cells. A: Representative immunofluorescence images of ODC-OVA slices incubated for 6 hours with in vitro activated OT-I T cells in the absence (left panel) and presence (right panel) of GA (50 μg/ml) in the hippocampus. (Inset) Nuclei of activated caspase-3+ neurons show inhomogeneous DAPI-staining consistent with nuclear condensation and fragmentation due to apoptotic cell death. DAPI (blue), NeuN (green), activated caspase-3 (red); scale bar represents 10 μm. C: Densities of activated caspase-3+ NeuN+ neuronal cell bodies detected in the hippocampal (upper panels) and cortical (lower panels) gray matter of acute brain slices from ODC-OVA mice incubated for 6 hours with (right panels) and without (left panels) in vitro activated OT-I T cells in the absence and presence of three different GA concentrations (10, 50, and 200 μg/ml). ns = not significant, **P < 0.05. B, D, and E: Neuronal (and oligodendroglial) apoptosis can be reduced by pre-incubation of OT-I splenocytes with GA before incubation in acute brain slices from ODC-OVA mice. B: Representative immunofluorescence images of ODC-OVA slices incubated for 6 hours with OT-I T cells in vitro activated in the absence (left panel) and presence (right panel) of GA (1 μg/ml). DAPI (blue), NeuN (green), activated caspase-3 (red). Scale bar = 10 μm. D: Densities of activated caspase-3+ NeuN+ neuronal cell bodies detected in the hippocampal (left panel) and cortical (right panel) gray matter of acute brain slices from ODC-OVA mice incubated for 6 hours without (left bar) or with OT-I T cells in vitro activated in the absence (middle bar) and presence (right bar) of GA (1 μg/ml). E: Densities of activated caspase-3+ NogoA+ oligodendrocytes detected in the hippocampal (left panel) and cortical (right panel) gray matter of acute brain slices from ODC-OVA mice incubated for 6 hours without (left bar) or with OT-I T cells in vitro activated in the absence (middle bar) and presence (right bar) of GA (1 μg/ml). ns = not significant, **P < 0.05.