Figure 3.

AAV vectors drive RGMa expression and secretion in the mouse brain. A, Quantification of the percentage of RGMa fluorescence intensity in SN and the striatum of the bilateral and unilateral high-titer-injected animals. RGMa protein levels were significantly increased in the injected SN compared with AAV-Empty-injected (p < 0.0001) and noninjected SN (p = 0.003). Similarly, striatal RGMa levels were significantly higher in the bilaterally AAV-RGMa-injected animals compared with the AAV-Empty-injected animals (p < 0.0001) and in the ipsilateral striatum versus the contralateral striatum of the unilaterally AAV-RGMa-injected animals (p = 0.0126). B, Quantification of RGMA fluorescence intensity in SN and striatum in low-titer AAV-RGMa-injected animals. The percentage of RGMa fluorescence intensity was significantly increased in the right SN injected with AAV-RGMa and, to a smaller extent, in the left noninjected SN, compared with all control groups (GFP, saline, or Empty, SN right, p < 0.0001, F_(3,25) = 159.0; SN left, p < 0.0001, F_(3,25) = 42.9). Both striatal (p = 0.001) and SN (p = 0.0001) levels of RGMa were significantly higher on the right transduced side of the nigrostriatal tract. Data are represented in percentages of fluorescence relative to the AAV-Empty RGMa fluorescence. For representative images of the RGMa immunohistochemical stained mouse SN and striatum, see Figures 6B, E and 7B, E. Tissue from 6 mice was measured in the high-titer treatment group, and from 7–8 mice in low-titer treatment group. Statistical analysis was performed using Student's t test and one-way ANOVA with Tukey's post hoc multiple testing correction: *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. All data are expressed as mean with bars representing SEM.