Fig. 7.
rADAMTS-4 reduces perineuronal nets enwrapping motoneurons in the lumbar spinal cord of SOD1G93A mice. a Representative photomicrographs of ventral horns in lumbar spinal cord sections from WT and control or ADAMTS-4-treated SOD1G93A male mice stained with WFA, a marker of perineuronal nets. Scale bar: 500 or 250 μm. b Quantification of WFA immunoreactivity per area from male mice (a). Values plotted are mean ± SEM. Two-way ANOVA: *** P < 0.001 WT Vs SOD1G93A. Unpaired two-tailed t-Test: $$ P < 0.01 Control Vs ADAMTS-4 SOD1G93A, N = 3 Control WT, N = 5 ADAMTS-4 WT, N = 8 Control SOD1G93A, N = 7 ADAMTS-4 SOD1G93A. c Representative photomicrographs of ventral horns in lumbar spinal cord sections from WT and control or ADAMTS-4-treated SOD1G93A female mice stained with WFA. Scale bar: 500 or 250 μm. d Quantification of WFA immunoreactivity per area from female mice (c). Values plotted are mean ± SEM. Two-way ANOVA: *** P < 0.001 WT Vs SOD1G93A. Unpaired two-tailed t-Test: $ P < 0.05 Control Vs ADAMTS-4 SOD1G93A, N = 5 Control WT, N = 3 ADAMTS-4 WT, N = 5 Control SOD1G93A, N = 6 ADAMTS-4 SOD1G93A. e Positive correlation between the percentage of WFA-positive perineuronal nets per area and the number of motoneurons in male and female WT (N = 15; blank diamond) and SOD1G93A (N = 24; black diamond) symptomatic mice from figs. 4–5. Spearman’s rank correlation: *** P < 0.001. R represents the coefficient of correlation. f-k Quantitative RT-PCR for aggrecan (f-g) HAPLN1 (h-i) and tenascin R (j-k) in the lumbar spinal cord of WT and SOD1G93A male (f, h, j) and female (g, i, k) mice at PS, SS and ES. Values plotted are mean ± SEM. Mann–Whitney U-tests: * P < 0.05 compared to corresponding WT mice, $ P < 0.05 compared to SOD1G93A mice at other stages, # P < 0.05 compared to WT mice at other stages, N = 3-4. l Immunoblot for CSPG (chondroitin sulfate proteoglycans) core proteins in lumbar spinal cord protein extracts of SOD1G93A mice exposed to human recombinant ADAMTS-1, ADAMTS-4 or ADAMTS-5 ex vivo for 24 h at 37 °C. Quantification of total CSPG core proteins. Values plotted are mean ± SEM. Mann–Whitney U-tests: * P < 0.05 between control and ADAMTS-4 conditions, N = 4