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. 2017 Mar 29;140(5):1447–1465. doi: 10.1093/brain/awx060

Figure 4.

Figure 4

AAV-Grn transduces neurons. Brains from AAV-treated mice were immunostained for progranulin to assess the spread of AAV-expressed progranulin (A). Strong progranulin immunoreactivity was detected throughout the mPFC of AAV-Grn-treated mice, as well as in other medial structures including striatum, septum, and ventromedial thalamus. Progranulin ELISA was performed to more quantitatively assess progranulin levels in mPFC and amygdala (B and C). Both wild-type and Grn+/– mice exhibited dramatic increases in progranulin levels in the mPFC (B, ANOVA main effect of virus, P < 0.0001), with a similar level of expression in each genotype. AAV-Grn-treated mice also had a modest, but statistically significant increase in progranulin in the amygdala, restoring progranulin levels to around normal in Grn+/– mice (C, ANOVA main effect of virus, P = 0.0001). In both regions, AAV-GFP-treated Grn+/– mice exhibited progranulin insufficiency as expected (B inset, C). Brains from AAV-treated mice were double-immunostained for the myc tag on virally-expressed progranulin and markers for neurons (D, NeuN), astrocytes (E, GFAP), or microglia (F, Iba1), revealing that of these three cell types, AAV-Grn only transduced neurons. Immunostaining and ELISA data were analysed by ANOVA. Tukey’s post hoc test was used for immunostaining and amygdala ELISA data. Genotype differences within viral group from the prefrontal cortex ELISA data were analysed by t-test due to the large difference in progranulin levels between AAV groups. Scale bars in representative double-labelled images represent 5 µm. n = 10–20 mice per group for progranulin immunostaining and 6–16 per group for progranulin ELISA. **P < 0.01, ***P < 0.001, ns = not significant.