From: Efficient Gene Transfer into Retinal Cells Using Adenoviral Vectors: Dependence on Receptor Expression Invest. Ophthalmol. Vis. Sci.. 2004;45(6):1680-1687. doi:10.1167/iovs.03-0730 Figure Legend: Comparison of transduction efficiency of AdV5-GFP to AdV5/F35-GFP in murine retina. Mice were given subretinal injections of either AdV5-GFP or AdV5/F35-GFP (107 IU). Enucleated eyes were examined 2 weeks after injection by immunohistochemistry using anti-GFP and peroxidase-conjugated second antibody as described in the Methods section. A sham-injected eye shows no staining of any retinal cell layers with anti-GFP (A). Retina pigment epithelium (RPE) stains reddish brown near the site of AdV5-GFP injection. These RPE cells appear enlarged and hyperplastic (B). GFP is also detected in Müller cells (MC) and occasional full-length cell bodies and nuclei of photoreceptors at the site of injection (C). Rag-2 and C57BL/6 mice examined 2 weeks after injection with AdV5/F35-GFP show reddish-brown staining in the photoreceptors and RPE beyond the injection sites (arrows). The retinal detachment is an artifact of processing as is the light brown staining in the lens (L) (D, E). Few nerve fibers in the optic nerve also stain (arrow) (F). Higher magnification indicates labeling of the entire photoreceptor including the outer segments (OS), inner segments (IS), nucleus (ONL), and synaptic zone in the outer plexiform layer (OPL) (G). Sections were also double labeled with anti-GFAP (stained brown), a glial cell marker, and anti-GFP (stained red). A Müller cell can be identified in the inner retinal layer that stains for both GFAP (brown) and GFP (red). The arrow points to a Müller cell foot process that stains positive for both chromogens. Cells containing large nuclei (ganglion cells) and the cells in the inner nuclear layer are negative for both GFAP and GFP (H). The green arrows point to brown-labeled glial cells in the ganglion cell layer (GC) that weakly stain red, suggesting the presence of GFP. Black arrows point to two cells in the inner nuclear layer (INL) showing the presence of GFP but not GFAP (I). These cells are possibly horizontal cells because they contain dendrites directed toward photoreceptors. The RPE shows transduced cells staining bright red and nontransduced cells with football-shaped brown melanin granules. The underlying choroid (C) contains dark brown-pigmented melanocytes (J). Magnification: (A, B) ×20; (C, G–J) ×40; (D, E) ×4; (F) ×10. Date of download: 10/25/2017 The Association for Research in Vision and Ophthalmology Copyright © 2017. All rights reserved.