Age-related changes in the retinal pigment epithelium (RPE).
Age-related changes in the retinal pigment epithelium (RPE).
复制标题
DOI：
10.1371/journal.pone.0038673
复制
发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Bonilha VL
中科院分区：
文献类型：
--
作者：
Gu X;Neric NJ;Crabb JS;Crabb JW;Bhattacharya SK;Rayborn ME;Hollyfield JG;Bonilha VL
视网膜中与年龄相关的变化通常伴随着视觉障碍，但其机制细节仍鲜为人知。蛋白质组学研究旨在更好地从分子水平了解视网膜色素上皮(RPE)的衰老机制。分离幼年(3~4月龄)和老年(24~25月龄)F344BN大鼠视网膜色素上皮(RPE)细胞，分成含有顶端微绒毛(MV)和RPE细胞体(CB)的亚细胞组。蛋白质经洗涤剂提取、SDS-PAGE分离、胰酶原位消化、LC-MS/MS分析，对幼年和老年大鼠RPE中检测到的精选蛋白进行免疫荧光和Western印迹分析。青年大鼠RPE MV共检测到356个蛋白质，老年大鼠RPE MV共检测到378个蛋白质，其中48%为各年龄组所共有。幼年大鼠RPE CB共鉴定出897种蛋白质，老年CB共鉴定出675种蛋白质，其中56%为各年龄组所共有。一些已鉴定的蛋白质，包括参与氧化应激反应的蛋白质，在RPE老化过程中总体丰度显示出数量和质量上的变化。首次在视网膜色素上皮细胞中鉴定出大量蛋白质。一种这样的蛋白质，Colltrin，定位于近端小管顶端刷状边缘的顶膜，在那里它可能调节几个氨基酸转运体。此外，胶原素还参与了胰腺β细胞的增殖和胰岛素的分泌。在RPE中，胶原蛋白的表达在RPE老化过程中显著调节。另一种受年龄调节的新蛋白质是DJ-1，这是一种在大脑中被广泛研究的蛋白质，其氧化应激相关功能已被描述。本文提供的数据揭示了RPE在老化过程中的具体变化，提供了第一个RPE老化的蛋白质数据库，这将有助于未来对与年龄相关的视网膜疾病的研究。
Age-related changes in the retina are often accompanied by visual impairment but their mechanistic details remain poorly understood. Proteomic studies were pursued toward a better molecular understanding of retinal pigment epithelium (RPE) aging mechanisms. RPE cells were isolated from young adults (3–4 month-old) and old (24–25 month-old) F344BN rats, and separated into subcellular fractions containing apical microvilli (MV) and RPE cell bodies (CB) lacking their apical microvilli. Proteins were extracted in detergent, separated by SDS-PAGE, digested in situ with trypsin and analyzed by LC MS/MS. Select proteins detected in young and old rat RPE were further studied using immunofluorescence and Western blot analysis. A total of 356 proteins were identified in RPE MV from young and 378 in RPE MV from old rats, 48% of which were common to each age group. A total of 897 proteins were identified in RPE CB from young rats and 675 in old CB, 56% of which were common to each age group. Several of the identified proteins, including proteins involved in response to oxidative stress, displayed both quantitative and qualitative changes in overall abundance during RPE aging. Numerous proteins were identified for the first time in the RPE. One such protein, collectrin, was localized to the apical membrane of apical brush border of proximal tubules where it likely regulates several amino acid transporters. Elsewhere, collectrin is involved in pancreatic β cell proliferation and insulin secretion. In the RPE, collectrin expression was significantly modulated during RPE aging. Another age-regulated, newly described protein was DJ-1, a protein extensively studied in brain where oxidative stress-related functions have been described. The data presented here reveals specific changes in the RPE during aging, providing the first protein database of RPE aging, which will facilitate future studies of age-related retinal diseases.
登录
查看更多内容
影响因子：
3.4
作者：
Bonilha, VL;Bhattacharya, SK;Crabb, JW
通讯作者：
Crabb, JW
影响因子：
4.4
作者：
Garcia, Gonzalo G.;Akha, Amir A. Sadighi;Miller, Richard A.
通讯作者：
Miller, Richard A.
影响因子：
64.8
作者：
Danilczyk, Ursula;Sarao, Renu;Penninger, Josef M.
通讯作者：
Penninger, Josef M.
影响因子：
2.9
作者：
Panahian, N;Huang, TJ;Maines, MD
通讯作者：
Maines, MD
影响因子：
2.9
作者：
DILORETO, DA;MARTZEN, MR;DELCERRO, M
通讯作者：
DELCERRO, M