Figure 5: Abnormal dendritic and synaptic development of newborn neurons with CRMP2 deficiency in the adult dentate gyrus.

(a) Confocal projection images of GFP⁺ neurons at 2 weeks post injection (w.p.i.) of retroviruses co-expressing GFP and control shRNA (Ctrl) or Crmp2 shRNA (shC2#2 and shC2#5). Scale bar, 20 μm. (b) Summary of total dendritic length of GFP⁺ newborn neurons expressing different shRNAs. Data were collected from 5, 3, 3 animals for cntl, shC2#2 and shC2#5, respectively. (c) Total dendritic length of GFP⁺ neurons at 2, 4 and 6 w.p.i. of retroviruses co-expressing GFP and either Ctrl or Crmp2 shRNA (shC2) in adult dentate gyrus. Data were collected from 5 (cntl)/3 (shC2), 3 (cntl)/5 (shC2), 3 (cntl)/4 (shC2) animals for 2, 4 and 6 w.p.i., respectively. (d–f) Sholl analysis at 2 (d), 4 (e) and 6 w.p.i. (f) with the same set of GFP⁺ neurons analysed in c. (g) Representative confocal projection images of dendritic spines from GFP⁺ neurons at 2, 4 and 6 w.p.i. Scale bar, 2 μm. Data were collected from 3 (cntl)/3 (shC2), 4 (cntl)/5 (shC2), 3 (cntl)/4 (shC2) animals for 2, 4 and 6 w.p.i., respectively. (h,i) Summary of total spine density (h) and mature spine density (i), with the same set of GFP⁺ neurons analysed in g. (j) Representative confocal projection images of mossy fibre boutons. Scale bar, 10 μm. (k) Summary of average volume of boutons. Data were collected from 5 cntl and 3 shC2 animals, respectively. Numbers associated with each bar graph refer to the total number of cells analysed under each condition. Values represent mean±s.e.m. (**P<0.01; ***P<0.001; (b,c,h,i) one-way ANOVA with Tukey post hoc test, (d–f) Kolmogorov–Smirnov test, (k) unpaired t-test).