Figure 1

Characterization of the effects of the intrastriatal delivery of AAV-GFP and AAV-hCDNF at 6 weeks. (A) Experimental scheme for analysing the transduction of AAV-GFP and AAV-hCDNF in the mouse brain. C57BL/6 mice received bilateral intrastriatal injections of AAV-GFP, AAV-hCDNF, or PBS as a control. Samples were collected at 2, 4 and 6 weeks after injection. (B) Immunohistochemical staining of striatum (STR) and substantia nigra (SN) sections for GFP (upper panel, brown) and CDNF (lower panel, brown) and counterstaining with cresyl violet (Nissl bodies). Scale bars: 500 µm for the STR, 200 µm for the SN, and 20 µm for the enlarged images. (C) Distribution profiles of GFP and hCDNF at 6 weeks after injection of AAV-GFP or AAV-hCDNF into the STR. (D) Western blot analysis of GFP and CDNF expression in the STR and SN. (E) Immunofluorescence staining of GFP (green) or hCDNF (green) in dopaminergic neurons (TH; magenta), microglia (Iba1; magenta) and astroglia (GFAP: magenta) with in the STR. Scale bar, 20 µm. (F) Double immunofluorescence staining of nuclei (DAPI, cyan) and GRP78 (red) with GFP (green) or hCDNF (green) in the SNpc. Scale bar, 20 µm. (G) Immunohistochemical staining of striatal sections for TH (upper) and DAT (lower). Scale bar, 1 mm, 200 μm for enlarged images of the STR. (H) Quantification of the TH⁺ or DAT⁺ fibre density in the STR. (I) Immunohistochemical staining of SN sections for TH. Scale bar, 200 µm. (J) Quantification of TH⁺ neurons in the SNpc. (C) n = 4 for the AAV-GFP group, n = 7 for the AAV-hCDNF group, (H,J) n = 5 for the PBS group, n = 5 for the AAV-GFP group, and n = 6 for the AAV-hCDNF group. (H,J) One-way ANOVA followed by Tukey’s multiple comparisons post hoc test. All the data are expressed as the mean ± S.E.M.