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. 2022 Jul 6;42(27):5346–5360. doi: 10.1523/JNEUROSCI.2258-21.2022

Figure 6.

Figure 6.

Depletion of the CA1 Egr1-dependent activated neurons during the rotarod task. A, Schematic representation of the double mutant mice used in this experiment. B, Schematic representation of the AAV-flex-taCasp3-TEVp vector. Mice were bilaterally injected in CA1 with AAV-flex-taCasp3-TEVp (Casp3, n = 8) or vehicle (Vehicle, n = 7). A group of NT mice (Naive, n = 5) was also used. C, All three groups received a single injection 4-HT 1 h before the rotarod training (arrows) on days 1 and 2 of training. Data are mean ± SEM, analyzed by two-way ANOVA. A significant difference was detected between groups on day 3, F(1,52) = 15.78, p = 0.0002; day 4, F(1,52) = 9.9588, p = 0.0027; and day 5, F(1,52) = 10.05, p = 0.0025. D, Representative images of Egr1-dependent activation of neural cells (GFP-positive, green) co-stained with DAPI (blue) in the dorsal CA1 of the hippocampus in the three groups. Scale bar: 60 µm. E, Quantification of hippocampal GFP-positive neural cells density per area in the three groups. Data are mean ± SEM. Two-way ANOVA identified general significant changes between groups, F(2,51) = 49.27, p < 0.0001. Tukey's post hoc analysis indicated that both, Naive (p < 0.05) and Casp3 (p < 0.001) showed significantly different GFP-positive cell density compared with Vehicle mice. *: Naive versus Vehicle groups; $: Casp3 versus Naive; &: Casp3 versus Vehicle. F, Representative images of Egr1-dependent activation of neural cells (GFP-positive, green) in the dorsal striatum (DStr) from the three groups. Scale bar: 120 µm. G, Quantification of striatal GFP-positive neural cells density per area in the three groups. H, A new cohort of double mutant mice as in A, were subjected to the same experimental design as in B, C but without 4-HT administration to rule out potential leakiness of the system. Data are mean ± SEM, analyzed by two-way ANOVA (no significant differences were detected between groups). I, Postmortem hippocampal examination in mice from H shows no recombination because of the absence of 4-HT administration. SO: stratum oriens, SP: stratum pyramidale, SR: stratum radiatum.