Viral vector delivery

AAV8–CaMKII–HA–rM₃D(G_s)–IRES–mCitrine (AAV–G_s–DREADD–mCitrine) or AAV8–CaMKII–GFP (AAV–GFP) viral vectors were generated by the University of North Carolina Viral Vector Core and infused into wild-type mice. Lentiviral vectors expressing GFP or Cre recombinase (Cre) under the CMV promoter were generated by the Emory University Viral Vector Core and infused into floxed Bdnf mice. Mice were anesthetized with ketamine/dexdomitor. With needles centered at bregma, stereotaxic coordinates were located on the leveled skull using a digitized stereotaxic frame. Viral vectors were delivered to +2.3 mm anteroposterior (AP), −2.8 mm dorsoventral (DV), and ±0.1 mm mediolateral (ML; Gourley et al., 2010) in a volume of 0.5 μl/side. The microsyringe remained in place for 5 min after infusion. Mice were sutured and allowed to recover for at least 3 weeks before behavioral testing. After testing, mice were deeply anesthetized and transcardially perfused with 4% paraformaldehyde, then brains were sectioned into 40 μm sections, and mCitrine or GFP was imaged to confirm that infusions infected the mOFC.