Treatment with FTY720 increases the number of DCX-positive cells in SGZ.
a, b Representative images acquired using confocal laser scanning microscopy of double immufluorescence staining for BrdU and DCX in ipsilateral DG in saline-injected control (vehicle), FTY720, KA and KA + FTY720 injected animals, 8 days after surgery. (i) Low magnification and (ii) Z-stack maximal projection of 5–10 confocal layers spaced by 2.5 μm of the area marked with a white frame in (i). Scale bar (i) 200 μm and (ii) 50 μm. c, d Quantitative analysis of BrdU+ and DCX+/BrdU+ cells within the SGZ of the ipsilateral dorsal hippocampus. Positive cells were counted using confocal acquired images over the total SGZ of the ipsi DG in two sections per animal. DCX and BrdU colocalization was determined examining three-dimensional orthogonal reconstruction of confocal layers (ImageJ software). Data are presented as c mean of positive cells per slice ± SEM and d percentage of DCX+ cells over BrdU+ total cells. Statistical analysis: one-way ANOVA followed by Bonferroni post hoc test; *p < 0.05, **p < 0.01. N = 6 for each experimental group. e Representative confocal Z-stack images showing BrdU and DCX colocalization, as single and merged channels