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Fig. 4 | Molecular Neurodegeneration

Fig. 4

From: Counteracting roles of MHCI and CD8+ T cells in the peripheral and central nervous system of ALS SOD1G93A mice

Fig. 4

MHCI depletion reduces the inflammation in the lumbar and cervical spinal cord of mSOD1 mice during the disease course. a, b DAB immunostaining for CD68 in the lumbar spinal cord of NTG; G93A+/+ and G93A−/− mice at 123 and 140 d. Bar, 50 μm. c Immunofluorescence staining for MHCI (purple) and motor neurons (neuro-trance, NT; blue) in the cervical spinal cord of NTG; G93A+/+ and G93A−/− mice at 140 d. The inset shows a magnification of MHCI-labeled microglia surrounding MNs. The images are representative of at least four sections from two independent experiments from each genotype. Bar, 50 μm; Inset Bar: 50 μm. d Immunofluorescence staining for CD68 in the cervical spinal cord of NTG; G93A+/+ and G93A−/− mice at 140 d. Bar, 50 μm (e, i) Quantification of CD68 staining in (e) lumbar and (i) cervical spinal cord hemisections of G93A+/+, G93A−/− mice compared to NTG+/+ littermates at 123 and 140 d. Data are expressed as mean ± SEM (four serial sections for each animal). At 123 d, four, six and four independent experiments were analyzed for NTG, G93A+/+ and, G93A−/− mice, respectively. At 140 d, in cervical and lumbar spinal cord, four independent experiments were analyzed for each genotype. f-h Real-time PCR for Ifn-γ, Ccl2 and Igf1 transcripts in the lumbar spinal cord of G93A+/+, G93A−/− mice compared to NTG+/+ littermates at 123 and 140 d. Data are normalized to β-actin and expressed as the mean ± SEM fold change ratio between G93A+/+ mice, G93A−/− mice and controls from four independent experiments for each genotype. j-l Real-time PCR for Ifn-γ, Ccl2 and Igf1 transcripts in the cervical spinal cord of G93A+/+, G93A−/− mice compared to NTG+/+ littermates at 140 d. Data are normalized to β-actin and expressed as the mean ± SEM fold change ratio between G93A+/+, G93A−/− and controls mice from four independent experiments for each genotype. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; (G93A+/+ vs G93A−/−); °P < 0.05; °°P < 0.01; °°°P < 0.001; °°°°P < 0.0001 (G93A−/−; G93A+/+ vs NTG) by one-way ANOVA with Tukey’s post-analysis

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