Yee Kenneth M P, Feener Edward P, Madigan Michele, Jackson Nicholas J, Gao Ben-Bo, Ross-Cisneros Fred N, Provis Jan, Aiello Lloyd Paul, Sadun Alfredo A, Sebag J
VMR Institute for Vitreous Macula Retina, Huntington Beach, California, United States 2Doheny Eye Institute, Los Angeles, California, United States.
Beetham Eye Institute, Joslin Diabetes Center, Boston, Massachusetts, United States.
Invest Ophthalmol Vis Sci. 2015 Nov;56(12):7036-42. doi: 10.1167/iovs.15-16809.
The proteomic profile of vitreous from second-trimester human embryos and young adults was characterized using mass spectrometry and analyzed for changes in protein levels that may relate to structural changes occurring during this time. This vitreous proteome was compared to previous reports to confirm proteins already identified and reveal novel ones.
Vitreous from 17 human embryos aged 14 to 20 weeks gestation (WG) and from a 12-, a 14-, a 15-, and a 28-year-old was individually analyzed using tandem mass spectrometry-based proteomics. Peptide spectral count associations with embryonic age were assessed using a general linear model of fold changes and Spearman's rank correlation. Differences between embryonic and young adult vitreous proteomes were also compared. Immunohistochemistry was used to evaluate three proteins in five additional fetal (10-18 WG) human eyes.
There were 1217 proteins identified in fetal and young adult human vitreous, 206 after quantile normalization and variance filtering. In embryos, the peptide counts of 37 proteins changed significantly from 14 to 20 WG: 75.7% increased, 24.3% decreased. Immunohistochemistry confirmed the absence of clusterin and cadherin in 10 and 14 WG eyes and their presence at 18 WG. Comparing embryonic to young adult vitreous, 47 proteins were significantly higher or lower. A total of 768 proteins not previously identified in the literature are presented.
Proteins previously unreported in the human vitreous were identified. The human vitreous proteome undergoes significant changes during embryogenesis and young adulthood. A number of protein levels change considerably during the second trimester, with the majority decreasing.
利用质谱分析法对孕中期人类胚胎和年轻人玻璃体的蛋白质组特征进行表征,并分析可能与该时期发生的结构变化相关的蛋白质水平变化。将该玻璃体蛋白质组与先前的报告进行比较,以确认已鉴定的蛋白质并揭示新的蛋白质。
使用基于串联质谱的蛋白质组学方法分别分析来自17例孕龄14至20周(WG)的人类胚胎以及一名12岁、一名14岁、一名15岁和一名28岁个体的玻璃体。使用倍数变化的一般线性模型和Spearman等级相关性评估肽谱计数与胚胎年龄的关联。还比较了胚胎和年轻成人玻璃体蛋白质组之间的差异。使用免疫组织化学方法评估另外五只胎儿(10 - 18 WG)人眼的三种蛋白质。
在胎儿和年轻成人的人类玻璃体中鉴定出1217种蛋白质,经过分位数归一化和方差过滤后为206种。在胚胎中,37种蛋白质的肽计数在14至20 WG之间有显著变化:75.7%增加,24.3%减少。免疫组织化学证实10和14 WG的眼睛中没有聚集素和钙黏蛋白,而在18 WG时存在。将胚胎玻璃体与年轻成人玻璃体进行比较,47种蛋白质显著更高或更低。总共呈现了768种先前文献中未鉴定的蛋白质。
鉴定出了先前在人类玻璃体中未报道的蛋白质。人类玻璃体蛋白质组在胚胎发育和年轻成年期经历了显著变化。在孕中期,许多蛋白质水平发生了相当大的变化,大多数呈下降趋势。