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Albarracin et al. BMC Neuroscience 2013, 14:125 http://www.biomedcentral.com/1471-2202/14/125 Page 5 of 14 (A) 0d NT 0d Tr 14 NT 14 Tr GCL INL ONL (B) 100 80 60 40 20 0 GCL INL ONL GCL INL ONL GCL INL ONL ** 0d 14d Figure 3 Effects of 670 nm light treatment on the population of photoreceptors hyperoxic mice. (A) Representative micrographs of retinal sections showing the nuclear layers stained with Bisbenzimide (blue). To account for obliquely cut retina and possible artefacts, the ratio of the ONL (white line) to the OLM-ILM (red line) was used for analyses. (B) Quantitative analysis of photoreceptor nuclei density in the inferior central region of NT group (black bar) and Tr mice (red bar). Compared with the nontreated retinas at 14d hyperoxia (14dNT), treatment with 670 nm red light induced a significant level of photoreceptor nuclei density preservation in the 14dTr group (*p < 0.001). Scale bar 50 μm in A. Nontreated 670nm Treated up-regulation of Hmox-1 expression in NT retinas, while in 10dTr retinas, levels of Hmox-1 expression remained at around 2-fold above baseline. Similar data were obtained from the 14d animals. The levels of Hmox-1 expression in the Tr animals were significantly less than in the NT animals at both 10d and 14d (p < 0.05). C3 Expression of C3 increased over longer periods of hyperoxia exposure (Figure 10B) in both Tr and NT animals. In the NT group (black bars), this increase above baseline was significant at 7d and 14d. At 10d and 14d, there were sig- nificantly higher levels of C3 expression in NT retinas (black bar) compared to Tr retinas (red bars) (p < 0.05). Fgf-2 A small and not statistically significant upregulation of Fgf-2 was detected at 3d in both Tr (Figure 10C, red bar) and NT animals (Figure 9C, black bar). After 7d hyperoxia, there was a 10-fold up-regulation of Fgf-2 in the NT retinas, compared with a 26-fold upregulation in Tr animals. Levels of expression were significantly differ- ent between 7dTr and 7dNT animals (p < 0.05). Levels of Fgf-2 continued to increase in NT animals at 10d and 14d; however, in Tr animals there was a fall in Fgf-2 expression at 10d to approximately 15-fold above baseline, which was maintained at 14d in the Tr group. At 10d and 14d, ex- pression levels of Fgf-2 in NT retinas were higher com- pared to Tr retinas, showing significant difference at 14d (p < 0.05). Discussion These findings corroborate earlier studies showing that hyperoxia causes oxidative stress, photoreceptor-specific cell death, and retinal inflammation [30,32,38]. Those stu- dies show that (i) hyperoxia-induced changes are evident Photoreceptor Nuclei Density (%) (Inferior Central “hotspot”)PDF Image | 670 nm light mitigates oxygen-induced degeneration
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