Wnt agonist treatment rescues ethanol-induced retinal cell differentiation defects.

<p>(A-C) Treatment with 350 nM GSK3β inhibitor (LSN 2105786) showed a rescue of Wnt+ cells in the peripheral CMZ, which were reduced after ethanol treatment. Control and ethanol treated embryos in this experiment appeared identical to those shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0201659#pone.0201659.g004" target="_blank">Fig 4A–4C</a> and were left out for brevity. (D) Quantification of Wnt+/Alcama+ and Wnt-/Alcama+ cells in the peripheral CMZ. (E-J) Treatment with LSN 2105786 on <i>Tg(TP1</i>:<i>GFP)</i> fish showed rescue of Notch signaling after LSN 2105786 treatment. Ethanol treated fish show reduced notch-activity (green) and photoreceptor terminal differentiation marker expression (zpr-1, red), which was restored by LSN 2105786 treatment. (K) Quantification of mCherry-positive, Notch-active cells per optic nerve containing optical section, in the retina showed a significant rescue of notch signaling after LSN 2105786 treatment. (L) Quantification of phosphoH3-positive cells in the retina showed a significant rescue of cell proliferation response after LSN 2105786 treatment at 72 hpf. (M) Quantification of total zpr-1 intensity in the ONL of the retina showed a significant decrease in photoreceptor marker expression after ethanol treatment (2–24 hpf) and subsequent rescue by LSN 2105786 treatment from 32–48 hpf. 100E+ LSN 2105786, and 150E+ LSN 2105786 indicates 100 mM and 150 mM ethanol exposed embryos treated with LSN 2105786, respectively. Images show rostral at top, lateral at left. Error bars indicate standard deviation. ‘**’ indicates statistical significance in comparison to control embryos (p<0.01). ‘*’ indicates statistical significance in comparison to control embryos (p<0.05). ‘a’ indicates statistical significance in comparison to 100 mM ethanol embryos (p<0.05). ‘b’ indicates statistical significance in comparison to 150 mM ethanol embryos (p<0.05). Scale bar = 10 μm for panels A-C. Scale bar = 10 μm for panels E-J.