Figure 3 of Nishiguchi, Mol Vis 2009; 15:187-199.


Figure 3. Retinal precursors were identified in the pars plana of adult rd1 mice. Double-headed arrows indicate the pars plana. A: At P30, many cells positive for rhodopsin as well as recoverin were seen in the pars plana of rd1 mice (n=39). B: The majority of the recoverin-positive cells were stained with PNA (n=12). C: The proportions of recoverin-positive cells in the pars plana at P30 that are positive for PNA or rhodopsin (mean±SEM) are presented. After the number of recoverin-positive cells within 320 µm width of the pars plan were determined from a single optical scan (10.0 μm thick), the number of those also positive for rhodopsin and PNA were determined from the same image. Three independent images randomly obtained from the same eye were analyzed to calculate the proportion of recoverin-positive cells that were also positive for rhodopsin (Rho) or PNA (PNA) or neither (Rcv) per animal. Average proportion (%) of cells for each category was determined from 6 animals. D: Many retinal precursors bridged both the retina and pars plana. Note that the cilioretinal border was sharply demarcated with DAPI staining (n=24). E: Recoverin-positive cells (arrowhead) in the pars plana at P120. Note that a gap between the pars plana and retina (open arrowhead) was seen with DAPI staining. F: Increased numbers of recoverin-positive cells were observed in the pars plana of P60 (31.2 fold; n=8) and P120 (7.6 fold; n=6) rd1 mice compared with P60 wild-type mice (n=6). The number of recoverin-positive cells within 320 µm width of the pars plan from a single optical scan (10.0 μm thick) were determined from three independent images randomly obtained from the same eye. Average number of recoverin-positive cells within 320 µm width of the pars plan (mean±SEM) were determined for each group (i.e., P60 wild-type mice, P60 rd1 mice, and P120 rd1 mice). Note that the number of recoverin-positive cells decreased with aging, from P60 to P120, in rd1 mice. G: A representative image of a single thick scan used for quantification of recoverin-positive (green) cells in the pars plana (filled arrowhead) is shown. A semitransparent white line outlines the cilioretinal border. Those that encompassed the border (open arrowhead) were excluded. Note that the immunopositive cells in the deep retina are defocused, while those in the ciliary epithelium of the pars plana appear sharper. The oval structures connected to a cell body through a thin process (filled arrow) were considered cone pedicles, thus excluded from the cell count. H, I, J: Recoverin-positive cells in the pars plana with synaptophysin-positive synaptic vesicles are presented (J; n=4; arrowhead). A merged image of H and I is shown. Scale bar equals 25 μm in D, E, G, and H and 50 μm in A and B. A, B, D, H, I, and J are thick scans (merged from 2 scans; each scan was 7.1 μm thick for A and B and 3.9 μm thick for D, G, H, and I, respectively). E is presented as a thin scan (a single scan 10.0 μm thick). Abbreviations: rhodopsin (Rho); recoverin (Rcv); ciliary body (CB); synaptophysin (Syn).