Neuritin up-regulates neurogenesis in the dentate gyrus of 7-week-old SD rats.

<p>(A) A experimental scheme is shown. 7-week-old male SD rats were injected with EGFP- or neuritin-lentiviral particles in the dentate gyrus of the hippocampus using a stereotaxic apparatus (n = 6 for each group). BrdU was intraperitoneally administered for 6 days. (B) After 2 weeks, GFP expression was confirmed by confocal laser scanning microscopy. The scale bars indicate 200 µm (low-scaled panel) and 50 µm (magnified panel). (C) Brain sections were stained with anti-GFP (green), anti-BrdU (red), and anti-NeuN (purple) antibodies. Orthogonal analysis of the staining was conducted by confocal microscopy. (D) Cells stained with BrdU antibody were counted. (E) Staining with anti-GFP and anti-BrdU antibodies was performed and counted. (F) Staining with anti-BrdU, anti-NeuN and anti-GFP antibodies was performed and counted. The results were normalized as percentage ratios to the staining in EGFP expressing rats. Rats with neuritin expression showed an increase in BrdU/GFP expressing cells by 330.77±130.78% compared to the rats with EGFP expression. Quantitative analysis of triple staining with BrdU/NeuN/GFP antibodies showed that neuritin expression induced a significant increase of newly generated neuronal cells in the dentate gyrus by 338.46±123.08% compared to EGFP-injection (*<i>p</i><0.05, Student's <i>t</i>-test).