Figure 4.
AAV2-CBA-eGFP- and AAV2-CBA-MIF-mediated neuronal transduction of GFP and MIF. A) Representative fluorescence micrographs from SD rat neuronal cultures showing GFP fluorescence (left), NeuN immunostaining (center), and merged images (GFP+NeuN; right) after incubation with AAV2-CBA-eGFP (1×109 viral gc/well) for 3 days (×40 view). B) Adult male SHRs were microinjected bilaterally into the PVN with AAV2-CBA-eGFP (0.5 μl; 1×109 gc). Four months later, rats were euthanized, and brains were removed and sectioned in preparation for detection of GFP and immunostaining with NeuN. Images are high-power (×40) fluorescence micrographs from the same cell field in the PVN, showing GFP fluorescence (left), NeuN immunostaining (center), and merged images (GFP+NeuN; right). Data are representative of 4 rats. C) Adult male SHRs were microinjected bilaterally into the PVN with AAV2-CBA-MIF (0.5 μl; 1×109 gc). Four months later, rats were euthanized, and brains were removed and sectioned in preparation for detection of MIF and NeuN via immunostaining. Images are high-power (×40) fluorescence micrographs from the field of cells outlined in the inset, showing MIF immunostaining (left), NeuN immunostaining (center), and merged images (MIF+NeuN; right). White arrows indicate MIF-expressing neurons; yellow arrows indicate MIF immunoreactivity within non-neuronal cells, probably glia. Insets: low-power (×10) fluorescence micrographs. 3rd v, third cerebroventricle. Data are representative of 6 rats. Scale bars = 50 μm; 100 μm (insets).