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Nadine von Thun und Hohenstein-Blaul, Oliver Gramlich, Maarten Ruitenberg, Andreas Gravius, Franz Grus; β-Amyloid deposition in human glaucomatous retinae revealed by proteomic and immunohistochemical analyses. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1139.
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Recently, Aβ has been reported to be implicated in the development of RGC apoptosis in glaucoma, with evidence of caspase-3-mediated abnormal APP processing and increased expression of Aβ in RGCs and optic nerves in experimental glaucoma. Additionally, decreased vitreous Aβ levels have been reported in patients with glaucoma, what is consistent with retinal Aβ deposition. Aim of this study was to develop and establish a method to analyze levels of β-Amyloid in human glaucomatous retinae (glaucoma, n=5) in comparison to healthy subjects (CTRL, n=5). Furthermore, it should be examined whether β-Amyloid is located intra- or extracellular in human RGCs.
Antibodies against APP, β-Amyloid/APP (4G8), β-Amyloid 1-40 and 1-42 were used in an antibody microarray approach to quantify β-Amyloid levels in the sample tissues by mean intensities (MI). Additionally, β-Amyloid was detected immunohistochemically with the same antibody setup in paraffin embedded eye tissues and levels were evaluated by mean values (MV) using a 0-3 grading system. For establishment, brain and eye samples from a transgenic AD mouse strain (tg2576) were used.
Both methods revealed up-regulated retinal β-Amyloid levels in AD mice, as well as in human glaucomatous retinae compared to age matched controls. In glaucomatous samples, levels of β-Amyloid 1-42 (FCA 3542) were significantly up-regulated (CTRL= 6878±2144 MI, glaucoma= 9815±4786 MI, p=0.027) and elevated levels were detected for anti-β-Amyloid/APP 4G8 (CTRL: 3066±1423 MI; glaucoma=4161±1786 MI, p=0.056). Semi-quantitative analysis of the immunohistochemical stainings in human retinae: mean values (MV) of β-Amyloid 1-40 and 1-42 (12F4) were significantly up-regulated in the glaucomatous group (anti-β-Amyloid 1-40: CTRL= 0.7±0.4 MV, glaucoma= 1.8±0.7 MV; p=0.001; anti-β-Amyloid 1-42 (12F4): CTRL= 0.6±0.5 MV, glaucoma= 1.8±1.2 MV; p=0.037), while β-Amyloid/APP showed only a trend toward up-regulation (4G8: CTRL= 1.3±0.8 MV, glaucoma= 1.8±0.7 MV; p=0.18) and APP did not differ. Moreover, β-Amyloid was visualized intracellular around the soma of RGC, as well as extracellular, presumably on single axons.
Antibody microarray and immunohistochemistry revealed an increase of M. Alzheimer specific β-Amyloid in glaucomatous subjects. Therefore, β-Amyloid might be involved in the pathology of glaucoma.
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