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. 2021 Jan 13;41(2):376–389. doi: 10.1523/JNEUROSCI.2142-20.2020

Figure 2.

Figure 2.

Increased neuronal activity does not prevent motor neuron death. A, Immunostaining of ChAT+ motor neurons (green) in L1 spinal cord sections from WT, SMA, and SMA + 4-AP mice with chronic intraperitoneal injections at P11. Scale bar: 100 µm. B, Quantification of L1 motor neuron counts of the same groups as in A at P1, P6, and P11. P1: WT (3) = 313 ± 16.09, SMA (3) = 278 ± 11.27, p = 0.149, two-tailed t test; P6: WT (3) = 314.7 ± 7.31, SMA (5) = 202.6 ± 10.84, SMA + 4-AP (3) = 195.7 ± 8.99, p for WT versus SMA = 0.0001, p for WT versus SMA + 4-AP = 0.0002, one-way ANOVA with Tukey's correction; P11: WT (4) = 314 ± 16.2, SMA (4) = 129.5 ± 17.81, SMA + 4-AP (6) = 141.7 ± 10.78, p for WT versus SMA < 0.0001, p for WT versus SMA + 4-AP < 0.0001, one-way ANOVA with Tukey's correction. n.s. = not significant. Number of mice is reported in parentheses. C, Immunostaining of ChAT+ motor neurons (green) in L2 spinal cord sections from the same groups as in A at P11. Scale bar: 100 µm. D, Quantification of L2 motor neuron counts of the same groups as in A at P11. WT (4) = 332 ± 6.22, SMA (4) = 263.9 ± 17.69, SMA + 4-AP (3) = 264.2 ± 10.31, p for WT versus SMA = 0.0107, p for WT versus SMA + 4-AP = 0.0164, one-way ANOVA with Tukey's correction. Number of mice is reported in parentheses. E, Immunostaining of ChAT+ motor neurons (green) and p53 or p-p53S18 (magenta) of L1 spinal cords from the same groups as in A at P11. Scale bar: 50 µm. F, G, Percentage of p53+ L1 motor neurons (F) and p-p53S18+ L1 motor neurons (G) in the same groups as in A. p53: WT (3) = 0 ± 0, SMA (3) = 64.60 ± 3.86, SMA + 4-AP (3) = 65.4 ± 3.85, p for WT versus SMA < 0.0001, p for WT versus SMA + 4-AP < 0.0001; p-p53: WT (3) = 0 ± 0, SMA (3) = 24.76 ± 4.46, SMA + 4-AP (3) = 21.93 ± 6.78, p for WT versus SMA = 0.0167, p for WT versus SMA + 4-AP = 0.0388, one-way ANOVA with Tukey's correction. Number of mice is reported in parentheses. Data are presented as mean ± SEM. Asterisks on top of bars without a horizontal line indicate the significance compared with the WT group; *p < 0.05, **p < 0.01, ***p < 0.001.