Fig. 2.

Increased Sema3D levels deteriorate cognitive functions. A Representative IHC stain of endogenous Sema3D protein in the brain of WT mice. Upper panel: whole-brain section; Lower panel: hippocampus. B-D Learning and spatial memory performance were examined by the MWM in control and Sema3D-overexpressing mice aged 4 months (n = 4 /group). Significant results from one-way repeated measures ANOVA for each type of mice as indicated by pound signs (#), and from two-way repeated measures ANOVA for comparisons between Sema3D-overexpressing and control mice as indicated by asterisk signs (*). B The control mice significantly improved their learning after 4-day practices but Sema3D-overexpressing mice failed to show significant improvement after 5-day practices. C Escape latency and D frequency were measured in the memory trials. These two types of mice had significant differences in memory over the 14-day test. E Sema3D-overexpressing mice significantly lost short-term memory on day 14 in the novel object recognition test (n = 4/group). F The Y-maze test was performed to measure spatial working memory in mice for 10 weeks (n = 14/each group). A lower percentage of alteration indicates worse spatial working memory. Data in Fig. 2B-F are presented as mean ± SEM. **p < 0.01 and ***p < 0.001. #p < 0.05 G The Y-maze test measuring spatial working memory was used to assess the effect of siRNA-Ctrl or siRNA-Sema3D injected into miR-195a KO mice aged 12 months (n = 7/each group). The tests for siRNA-Sema3D and siRNA-Ctrl mice were conducted on the same experiment day. Two-way repeated measures ANOVA showed siRNA-Sema3D significantly improved spatial memory (*p < 0.05). H Locomotor function was determined using the total travel distance, with longer distance indicating better locomotor function