Figure 6.

GluN2 and GFP transgene expression in vivo mediated by adeno-associated virus. In this experiment, AAV viruses designed to express Flag-GluN2A/B, GFP, and Flag-GluN2A/BΔC from a CMV, EFS, TRE3G or 0.4αCaMKII promoter, as indicated, were infused into the basolateral complex of the amygdala (BLA). All viruses were infused into the rat BLA, except for the TRE3G promoter containing viruses – these viruses were infused into the BLA of αCaMKII-tTA transgenic mice. Twenty one days following viral infusion, coronal sections were prepared that contained the BLA and native GFP expression was observed via fluorescence microscopy and Flag-GluN2 expression was observed via immunohistochemistry(IHC) and fluorescence microscopy. Images depict DAPI stained nuclei with the same fields viewed for GFP or Flag-GluN2 (Texas Red) transgene expression. Viruses designed to express GFP and Flag-GluN2A/BΔC exhibited convincing and robust transgene expression in vivo. Viruses designed to express Flag-GluN2A/B were not capable of conferring GluN2 expression in vivo. *indicates virus derived from pAAV-Basic vector. Coronal sections from naïve controls were processed as negative controls for Flag-IHC and coronal sections from wild type mice were processed as controls (Control A) for mice infused with AAV-TRE3G-Flag-GluN2A.