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Albarracin et al. BMC Neuroscience 2013, 14:125 Page 6 of 14 http://www.biomedcentral.com/1471-2202/14/125 INL0d 3d 7d 10d 14d BBZ TUNEL ONL Figure 4 Effects of 670 nm light pretreatment on photoreceptor cell death following hyperoxia. Representative images of TUNEL-stained sections sampled from the inferior region of the NT and Tr retinas exposed to hyperoxia at different timepoints. TUNEL + labelling (red) was undetectable in the NT and Tr retinas from 0d (controls). A small population of TUNEL + photoreceptors was present in 3dNT retinas and continued to increase in number from 7, 10 and 14 days of exposure to hyperoxia. TUNEL + cells are also present in the 670 nm light-treated groups with significantly reduced number. The blue staining is Bisbenzimide, a DNA-specific stain, identifying the nuclear layers of the retina. Scale bar 25 μm. (A) 100 100 75 75 50 50 25 25 00 IP IC ON SC SP (B) 0d NT 0d Tr IP IC ON SC SP Area 3dNT Ave# TUNE+L SEM 3d Tr Ave# TUNE+L SEM P Value n=8 Area 10d NT Ave# TUNE+L SEM 10dTr Ave# TUNE+L SEM P Value n=8 IP 16.7 +2.5 1 +0.4 ns IP 8.8 +1.9 12.9 +3.6 ns IC 30.7 +3.3 12.3 +3.7 *P<0.001 IC 77.1 +9.2 22.1 +1.9 *P<0.001 SC 2 +0.2 2.8 +0.8 ns SC 16.6 +4.9 3.9 +2.0 ns SP 0.3 +0.1 0.3 +0.3 ns SP 3.3 +2.4 0 +0 ns IP 7dNT 6.3 +3.0 7d Tr 1 +0.4 ns IP 14d NT 43.4 +4.9 14dTr 32.9 +2.3 ns IC 45.0 +1.4 12.3 +3.7 *P<0.001 IC 77.0 +5.3 35.9 +5.5 * P<0.001 SC 9.8 +5.1 2.8 +0.8 ns SC 39.8 +8.4 18.5 +5.6 *P<0.01 SP 2.0 +0.2 0.25 +0.1 ns SP 36.6 +6.4 14.5 +0.7 *P<0.001 Figure 5 Quantitative analyses of photoreceptors death. (A) TUNEL + photoreceptor cells were counted across the retina (from the inferior to superior) in 0d, 3d, 7d, 10d and 14d animals and data from Nontreated (solid colored lines) and 670 nm Treated (broken colored lines) groups were compared. TUNEL + cells were present in the inferior central area (hotspot) of the nontreated retinas from 3d (solid red line) and the number of cell death increased as a function of hyperoxia-exposure time. (B) In the 14dNT retinas (solid green line), hyperoxia exposure caused widespread cell death that also affected the photoreceptors in the superior retina. By contrast, these numbers were significantly reduced by 670 nm light pretreatment. *Statistically significant difference (p < 0.001) compared to NT groups. Treated Nontreated Ave. No. of TUNEL+ cells Ave. No. of TUNEL+ cellsPDF Image | 670 nm light mitigates oxygen-induced degeneration 2013
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