Figure 3

ISO-1 treatment blocks photoreceptor apoptosis and loss of ONL thickness. Experimental design (a): ISO-1(40 mg/kg/day) or vehicle treatment was initiated 1 day prior to RD creation and continued daily until sacrifice at day 3 or 14 (n = 6/group). Representative photographs show TUNEL positive cells (red) in the ONL in day 3 detached retina from vehicle (b) and ISO-1 treated mice (c). Scale bar denotes 50 microns. TUNEL positive cells were significantly reduced in ISO-1 treated mice at day 3 when apoptosis is maximal (f, *p = 0.0036), but not different at day 14 (not shown). A second independent study confirmed the significant reduction in TUNEL by ISO-1 (35 mg/kg/day, n = 6/group, p = 0.008, not shown). ONL thickness in ISO-1 treated eyes was significantly thicker than that of vehicle treated eyes at day 14 (g, *p = 0.0278). Confocal micrographs shows activated caspase-3 positive cells (green) in the ONL of day 3 background controls (d) and ISO-1 treated (e) mice (n = 10/group). Scale bar denotes 25 microns. Caspase-3 positive cells were not significantly reduced in ISO-1 treated animals compared to controls in day 3 and 14 (h, n = 10/group, p = 0.2593 and p = 0.1367, respectively). Cleaved PARP protein expression in ISO-1 treated eyes was not significantly different from vehicle treated eyes at either timepoints (i, n = 6, p = 0.3269 at day3 and n = 5, p = 0.3163 at day14). AIF protein expression in ISO-1 treated eyes was not significantly increased from vehicle treated eyes at day 3 (j, n = 6/group, p = 0.2891).