Elevated Albumin in Retinas of Monkeys with Experimental Glaucoma
Elevated Albumin in Retinas of Monkeys with Experimental Glaucoma
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DOI：
10.1167/iovs.09-4331
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发表时间：
2010-02-01
影响因子：
4.4
通讯作者：
Redell, John B.
中科院分区：
文献类型：
--
作者：
Carter-Dawson, Louvenia;Zhang, Yujin;Redell, John B.
目的.为了确定一个突出的蛋白质，约70 kDa，这是显着增加，在实验性青光眼的猴子视网膜相比，同胞控制视网膜，青光眼的严重程度的关系，其在视网膜中的定位。视网膜提取物进行二维凝胶电泳，以确定差异表达的蛋白质。通过液相色谱/质谱/质谱（LC/MS/MS）分离和碰撞诱导解离测序分析和鉴定来自丰富的70 kDa蛋白的纯化肽。在MASCOT（Matrix Science，Boston，MA）上进行蛋白质鉴定，并通过蛋白质印迹确认。通过回归分析研究了这种蛋白的增加与青光眼严重程度之间的关系。蛋白质定位在视网膜进行了评估，免疫组化与共聚焦成像。从8个不重叠的蛋白质片段中鉴定出丰富的蛋白质为猕猴血清白蛋白前体（67 kDa），并通过Western blot证实了其身份。视网膜白蛋白含量平均增加2.3倍（P = 0.015）。在青光眼眼，白蛋白定位于内核层、内丛状层和沿着玻璃体表面的一些神经元，而在对照组视网膜中仅在血管中发现。白蛋白是在患有青光眼的猴子视网膜中发现的丰富蛋白质。增加的白蛋白主要定位于视网膜内层，已知与实验性青光眼相关的氧化损伤在此处是突出的。由于白蛋白是一种主要的抗氧化剂，实验性青光眼患者视网膜中白蛋白的增加可能有助于保护视网膜免受氧化损伤。（Invest Ophthalmol维斯科学。2010; 51：952-959）DOI：10.1167/iovs.09-4331
PURPOSE. To establish the identity of a prominent protein, approximately 70 kDa, that is markedly increased in the retina of monkeys with experimental glaucoma compared with the fellow control retina, the relationship to glaucoma severity, and its localization in the retina.METHODS. Retinal extracts were subjected to 2-D gel electrophoresis to identify differentially expressed proteins. Purified peptides from the abundant 70 kDa protein were analyzed and identified by liquid chromatography/mass spectrometry/mass spectrometry (LC/MS/MS) separation, and collision-induced dissociation sequencing. Protein identity was performed on MASCOT (Matrix Science, Boston, MA) and confirmed by Western blot. The relationship between the increase in this protein and glaucoma severity was investigated by regression analyses. Protein localization in retina was evaluated by immunohistochemistry with confocal imaging.RESULTS. The abundant protein was identified as Macaca mulatta serum albumin precursor (67 kDa) from eight non-overlapping proteolytic fragments, and the identity was confirmed by Western blot. The average increase in retinal albumin content was 2.3 fold (P = 0.015). In glaucoma eyes, albumin was localized to some neurons of the inner nuclear layer, in the inner plexiform layer, and along the vitreal surface, but it was only found in blood vessels in control retinas.CONCLUSIONS. Albumin is the abundant protein found in the glaucomatous monkey retinas. The increased albumin is primarily localized to the inner retina where oxidative damage associated with experimental glaucoma is known to be prominent. Since albumin is a major antioxidant, the increase of albumin in the retinas of eyes with experimental glaucoma may serve to protect the retina against oxidative damage. (Invest Ophthalmol Vis Sci. 2010; 51: 952-959) DOI:10.1167/iovs.09-4331