Fig. 2

Reduced dendritic spine density in neurons lacking DGCR2. A-B Reduced spine density by DGCR2 knockdown in primary cultured hippocampal neurons. Representative images of neuronal morphology (upper panel) and dendritic spines (lower panel) of cultured hippocampal neurons (A). Neurons were isolated from rat E18.5 embryos, transfected at DIV9 with indicated constructs, and fixed for staining at DIV16. Sh-control, empty pSUPER-vector. Scale bars as indicated. Spine density (per 10 μm) quantitative analysis of data in A (B). n = 15 neurons for each condition. ** p < 0.005, One-way ANOVA. C-D Reduced spine density by DGCR2 knockdown in the hippocampus in vivo. Representative images of dendritic spines of electroporated hippocampal CA1 neurons (C). In-utero electroporation of DGCR2 shRNA (sh-540) and its control scramble shRNA (sh-540-scr) to the hippocampi of E14.5 or E15.5 embryos. At P30 after birth, sections were stained with anti-GFP antibody and subjected to spine analysis. Scale bars as indicated. Spine density quantitative analysis of data in C (D). n = 30 neurons for each condition. ** p < 0.005, Student’s t test. E-F Reduced spine density in Dgcr2 mt mice. Representative spine images from Golgi staining (E). Dendrite segments were chosen from hippocampal CA1 pyramidal neurons. Scale bars as indicated. Spine density quantitative analysis of data in E (F). n = 35 neurons for wt and n = 40 neurons for mt. ** p < 0.005, Student’s t test