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Albarracin et al. BMC Neuroscience 2013, 14:125 http://www.biomedcentral.com/1471-2202/14/125 Page 7 of 14 death induced by oxidative stress. The data demonstrate that pretreatment with 9 J/cm2 of 670 nm once daily for 5 consecutive days in this model: (i) delays the onset, and slows the progression of photoreceptor death, and (ii) downregulates the associated oxidative stress mar- kers, acrolein and Hmox-1. This decrease in oxidative stress leads to the attenuation of damage to the retina and the reduction of expression of proinflammatory C3 expression and reduced expression of the stress- inducible neuroprotectant, Fgf-2. Significantly, treatment of 670 nm light alone did not cause measurable damage in non-challenged retinas. 670 nm light treatment promotes photoreceptor survival The timecourse of photoreceptor degeneration in this model has been reported previously [30]. In the present study, we find that the severity of the hyperoxia-induced damage in photoreceptors is significantly reduced, but not prevented, by pretreatment with 670 nm red light. We used two methods to quantify damage to photoreceptors: measures of ONL thickness, and counts of numbers of apoptotic (TUNEL positive) photoreceptors. Analyses of ONL thickness, the cumulative measure of cell loss, indi- cates that 14dTr animals have an ONL thickness profile 225 200 175 150 125 100 75 50 25 0 Figure 6 Quantitative analysis of the average total number of TUNEL + cells. Total counts of TUNEL + photoreceptors across the entire section of NT (black bar) and Tr (red bar) retinas from 0d, 3d, 7d, 10d and 14d groups. *Statistically significant difference (p < 0.05) between groups marked with black or red lines. Nontreated 670nm Treated 0d 3d 7d 10d 14d Days exposure in hyperoxic environment (75% oxygen) by 3d exposure to hyperoxia, (ii) that there is a focal point of pathology in the inferior central retina, and (iii) this pathology spreads to other regions of the retina over time. In addition, the present findings show that pretreat- ment with 670 nm light effectively offsets photoreceptor Figure 7 Immunohistochemical analysis of Cox Va and acrolein. (A-F) Effects of 670 nm light treatment on the metabolic status and oxidative stress profiles of the mouse retinas following exposure to 10d and 14d of hyperoxic exposure were examined by immunohistochemical staining. Cryosectioned retinas from the inferior region of the NT and Tr groups were labelled with marker for mitochondrial metabolism, Cox Va (green) and indicator of oxidative stress, acrolein (red). Compared to 0d control retina (A), Cox Va labelling patterns (green) in the 10dNT and 14dNT were disrupted (B-C, green) but not in the 10dTr and 14dTr groups (E-F, green). Exposure to hyperoxia caused a significant increase in the acrolein expression in 10dNT and 14dNT retinas (B-C, red) compared to 0d control (D). The acrolein labelling was also present in the 10dTr and 14dTr groups but the intensity was much lower than the NT retinas (E-F, red). Scale bar 25 μm. Ave. Number of TUNEL+ cells per sectionPDF Image | 670 nm light mitigates oxygen-induced degeneration
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