Journal: Molecular Therapy. Methods & Clinical Development
Article Title: Comparison of AAV-Mediated Optogenetic Vision Restoration between Retinal Ganglion Cell Expression and ON Bipolar Cell Targeting
doi: 10.1016/j.omtm.2020.05.009
Figure Lengend Snippet: Expression and Specificity of CoChR-GFP following ON-BC Targeting (A–L) Representative immunofluorescence images show the triple labeling of GFP (green), G γ13 (purple), and PKC (red) in retinal vertical sections (A–F) and retinal whole mounts (G–L). G γ13 and PKC are ON-BC- and RBC-specific markers, respectively. Images show single labeling of GFP (A and G), G γ13 (B and H), and PKC (D and J); double labeling of GFP/G γ13 (C and I) and GFP/PKC (E and K); and triple labelign of GFP/G γ13 /PKC (F and L). As evidenced by the co-labeling of GFP/ G γ13 in retinal vertical sections, the expression of CoChR-GFP was almost exclusively targeted to ON-BCs in the inner nuclear layer (C). The majority of ON-BCs and RBCs were transduced by virus vectors based on the co-labeling of GFP/G γ13 (C and I) and GFP/PKC (E and K), respectively. The images of retinal whole mounts were captured with the focal plane in the inner nuclear layer. INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer. The images were captured from TKO mice. Scale bar, 50 μm.
Article Snippet: The following primary antibodies were used in this study: mouse anti-GFP (1:1000; Neuromab, UC Davis, Davis, CA, USA), mouse anti-PKC (1:10,000; Santa Cruz, Dallas, TX, USA), rabbit anti-G γ13 (1:1,000; sc-368324, Santa Cruz, Dallas, TX, USA), and rabbit anti-RBPMS (1:1,000; ABN1362, MilliporeSigma, Temecula, CA, USA).
Techniques: Expressing, Immunofluorescence, Labeling, Virus