FIGURE 5

FIGURE 5

Competence factor of red cone precursor cells, thrb, is upregulated in the light-damaged retina. (Aa–aj) Single z-plane confocal images of retinal sections from light-damaged Tg[thrb:Tomato]q22 zebrafish (Ag–l,As–aj), (0, 48, 60, 72, 84, 96 hLT) immunolabeled for PCNA (Aa–f,As–ad) and the red/green double cone marker, Zpr-1 (Am–x,Aae–aj) and counterstained with DAPI (As–x). (Ay–aj) Regions outlined in panels (As–x) at higher magnification. Arrows, thrb:Tomato-positive INL cells. Scale bars, 20 μm (Aa) and 10 μm (Ay). (B,C) Number of PCNA-positive, thrb:Tomato-positive and PCNA and thrb:Tomato-double positive cells in the INL (B) and ONL (C) over the light treatment timecourse. (D) Number of thrb:Tomato-positive and thrb:Tomato and PCNA-double positive cells in the INL at a different scale to (B). (E) Number of thrb:Tomato-positive, thrb:Tomato and PCNA-double positive cells in comparison to Zpr-1-positive cells and thrb:Tomato, PCNA and Zpr-1-triple positive cells in the ONL of retinas exposed to constant intense light for 0, 36, 48, 60, 72, 84 and 96 h. Mean ± SE, n ≥ 9, ^∗p_Tukey < 0.05 and ^#p < 0.05 indicate comparisons to 0 hLT for the different measures that were assessed. The symbols are color-coded according to the line that they represent in the corresponding graphs (p_ANOVA, see Table 1). Note, significance was not determined for PCNA in panels (B,C) and symbols indicating significance for thrb:Tomato-positive and thrb:Tomato and PCNA-double-positive cells are not shown in panel (E), as they are indicated in panel (C). (F) Single z-plane confocal images of retinal sections from Tg[thrb:Tomato]q22 zebrafish (Fa–d,g,h) co-labeled with Zpr-1 (Fe–h) and DAPI (Fa,b) at 96 hLT [cells indicated by arrowhead in panel (Aad) at higher zoom] and 2 drec. Arrowheads illustrate the presence of small inner/outer segments. Astericks, thrb:Tomato and Zpr-1 double positive cells. Arrows, Zpr-1-positive and thrb:Tomato-negative cells. Scale bar, 10 μm.