Fig. 7.

Purinergic signalling effect on lin28a and ascl1a transcriptional expression in the zebrafish retina. Total RNA was purified from pools of 10–16 neural retinas from which the peripheral tissue had been carefully removed (mature retina without the CMZ) (a, b). Each retina was obtained from uninjured eyes, which had been in vivo treated for 3 days with saline solution or 3 μM ADPβS (a slowly hydrolysable ADP analogue) solution injected within the vitreous cavity. Retina pools were obtained from three independent assays of ADPβS-treated uninjured retinas (graphical bars labelled with ADPβS-1, ADPβS-2, and ADPβS-3 tags) which were considered samples of interest. Fold change represents the relative expression ratio (rER) calculated and plotted separately for each independent assay. Calibrator samples were three independent pools of saline solution-treated uninjured retinas (fold change for each of the three control retina pools was equal to 1 and are graphically represented in the same control bar). c, d RNA was purified from pools of 10–16 retinas (mature portion only without the CMZ), each obtained from control saline-, 6 μM ouabain plus saline-, or ouabain plus 1 μM MRS2179-injected eyes. Intravitreous injections of MRS2179 were performed once daily from 0 to 48 h after lesion (hpl), and retinas were isolated 65 hpl. Fold change represents the average rER calculated from each of three independent retinal pools treated with ouabain or ouabain plus MRS2179. The calibrator sample was calculated as the average of three independent pools of saline solution-treated uninjured retinas. Data were expressed as mean ± SE. ***p < 0.001; **p < 0.01; *p < 0.05, in Tukey’s multiple comparison test after ANOVA