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. 2018 May 31;19(6):1636. doi: 10.3390/ijms19061636

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© 2018 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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(A) Exemplary retinal cross-sections of the four groups stained with anti-Brn-3a for RGCs (green), Bax for apoptotic cells (red), and DAPI for cell nuclei (blue). Fewer Brn-3a⁺ RGCs and more Bax⁺ apoptotic cells were noted in ischemic retinae; (B) A significant loss of Brn-3a⁺ ganglion cells was revealed in the ischemic (p = 0.02) and the bevacizumab group (p = 0.04), but not in ranibizumab-treated retinae (p = 0.35); (C) Significantly more apoptotic cells could be observed in ischemic eyes (p = 0.03), but not in the groups treated with bevacizumab (p = 0.22) and ranibizumab (p = 0.57); (D) A significant reduction of relative Brn-3a (Pou4f1) mRNA expression was measured in all ischemic groups via qRT-PCR. The untreated (p = 0.023), the bevacizumab- (p = 0.018), and ranibizumab-treated eyes (p = 0.017) expressed a lower Brn-3a (Pou4f1) mRNA level than the controls; (E) The relative Bad mRNA expression was significantly elevated in ischemic (p = 0.002) eyes. No effect in Bad mRNA expression could be detected between the control and the bevacizumab- (p = 0.131) and ranibizumab-treated groups (p = 0.491); (F) Regarding the relative caspase-3 mRNA expression, a significant up-regulation was noted in ischemic (p = 0.026), bevacizumab (p < 0.001), and ranibizumab-treated retinae (p = 0.009), when compared to the control group; (G) Analysis of relative caspase-9 mRNA expression revealed no differences in the expression level between the control and all ischemic eyes (ischemia: p = 0.131, beva: p = 0.118, rani: p = 0.858); (H) A significant elevation of NF-κB mRNA levels was detected in the ischemic (p = 0.034) and bevacizumab-treated group (p = 0.008) in comparison to control retinae. No differences in relative NF-κB mRNA expression were seen between the control and ranibizumab-treated retinae (p = 0.069). *: p < 0.05; **: p < 0.01; ***: p < 0.001. Abbreviations: GCL: ganglion cell layer, IPL: inner plexiform layer, Beva: bevacizumab, Rani: ranibizumab. Arrows: colocalization. Scale bar: 20 µm. Immunohistology: n = 5–6/group; qRT-PCR: n = 4/group.

(A) Exemplary retinal cross-sections of the four groups stained with anti-Brn-3a for RGCs (green), Bax for apoptotic cells (red), and DAPI for cell nuclei (blue). Fewer Brn-3a⁺ RGCs and more Bax⁺ apoptotic cells were noted in ischemic retinae; (B) A significant loss of Brn-3a⁺ ganglion cells was revealed in the ischemic (p = 0.02) and the bevacizumab group (p = 0.04), but not in ranibizumab-treated retinae (p = 0.35); (C) Significantly more apoptotic cells could be observed in ischemic eyes (p = 0.03), but not in the groups treated with bevacizumab (p = 0.22) and ranibizumab (p = 0.57); (D) A significant reduction of relative Brn-3a (Pou4f1) mRNA expression was measured in all ischemic groups via qRT-PCR. The untreated (p = 0.023), the bevacizumab- (p = 0.018), and ranibizumab-treated eyes (p = 0.017) expressed a lower Brn-3a (Pou4f1) mRNA level than the controls; (E) The relative Bad mRNA expression was significantly elevated in ischemic (p = 0.002) eyes. No effect in Bad mRNA expression could be detected between the control and the bevacizumab- (p = 0.131) and ranibizumab-treated groups (p = 0.491); (F) Regarding the relative caspase-3 mRNA expression, a significant up-regulation was noted in ischemic (p = 0.026), bevacizumab (p < 0.001), and ranibizumab-treated retinae (p = 0.009), when compared to the control group; (G) Analysis of relative caspase-9 mRNA expression revealed no differences in the expression level between the control and all ischemic eyes (ischemia: p = 0.131, beva: p = 0.118, rani: p = 0.858); (H) A significant elevation of NF-κB mRNA levels was detected in the ischemic (p = 0.034) and bevacizumab-treated group (p = 0.008) in comparison to control retinae. No differences in relative NF-κB mRNA expression were seen between the control and ranibizumab-treated retinae (p = 0.069). *: p < 0.05; **: p < 0.01; ***: p < 0.001. Abbreviations: GCL: ganglion cell layer, IPL: inner plexiform layer, Beva: bevacizumab, Rani: ranibizumab. Arrows: colocalization. Scale bar: 20 µm. Immunohistology: n = 5–6/group; qRT-PCR: n = 4/group.