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. 2019 Oct 30;12:260. doi: 10.3389/fnmol.2019.00260

Figure 5.

Figure 5

Knockdown of Ezh2 rescues the deficits associated with the loss of miR-137. (A) Diagram showing that lentivirus encoding shRNA targeting Ezh2 (shEzh2) or negative control (Scramble) were injected into the dentate gyrus (DG). (B) The shEzh2 lentivirus could efficiently knock down the expression of EZH2 protein in the hippocampus. (C,D) In the light/dark box test, shEzh2 mice demonstrated improved time (C) and entries (D) in light box (cKO + shEZH2 vs. cKO + Scramble: n = 12–14 mice per group, **p < 0.01). (E,F) In the open field test, shEzh2 reduced the anxious levels in miR-137 cKO mice, as indicated by significantly increased time (E) spent in the central zone (cKO + shEZH2 vs. cKO + Scramble: n = 12–14 mice per group, *p < 0.05) and more entries (F) in it (cKO + shEZH2 vs. cKO + Scramble: n = 12–14 mice per group, *p < 0.05). (G) sh-EZH2 dramatically reduced dendritic complexity in miR-137 deletion neurons compared with controls, as determined by Sholl analysis (n = 35–42 neurons per group, ***p < 0.01, ***p < 0.001). (H) sh-EZH2 significantly reduced dendritic length (n = 35–42 neurons per group, **p < 0.01, ***p < 0.001). (I) sh-EZH2 had a significant effect on the number of dendritic branches (n = 35–42 neurons per group, *p < 0.05, **p < 0.01). (J) Knockdown of EZH2 in miR-137 cKO neurons reduced the spine density to a degree similar to that seen in WT group (n = 38–40 neurons per group, **p < 0.01, ***p < 0.001). Left: representative images of dendritic spines. Right: statistical analyses of spine density. (K) sh-EZH2 rescued the impaired LTP in miR-137 cKO mice. Left: pooled time-course data of LTP from all recordings made from miR-137 cKO + scramble, miR-137 cKO + shEZH2, and miR-137 WT + scramble mice. Right: average LTP amplitude measured 55–60 min post-induction (n = 5–6 slices from four mice per group, **p < 0.01). n.s., non-significant.