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Combined retinal proteome datasets in response to atropine treatment using iTRAQ and SWATH-MS based proteomics approaches in guinea pig myopia model

机译:结合视网膜蛋白质组数据集响应于使用ITRAQ和基于SWATH-MS的蛋白质组学方法在豚鼠近视模型中的蛋白质组学方法

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摘要

Atropine, a non-selective muscarinic antagonist, is known to slow down myopia progression in human adolescents and in several animal models. However, its underlying molecular mechanism is unclear. The present work built a monocular form-deprivation myopia (FDM) guinea pig model, using facemasks as well as atropine treatment on FDM eyes for 2 and 4 weeks. Retinal protein changes in response to the FDM and effects of topical administration of atropine were screened for the two periods using fractionated isobaric tags for a relative and absolute quantification (iTRAQ) approach coupled with nano-liquid chromatography-tandem mass spectrometry (nano-LC–MS/MS) (n=24, 48 eyes). Retinal tissues from another cohort receiving 4-weeks FDM with atropine treatment (n=12, 24 eyes) with more significant changes were subjected to sequential window acquisition of all theoretical mass spectra (SWATH-MS) proteomics for further protein target confirmation. A total of 1695 proteins (8875 peptides) and 5961 proteins (51871 peptides) were identified using iTRAQ and SWATH approaches, respectively. Using the Paragon algorithm in the ProteinPilotTM software, the three most significantly up-regulated and down-regulated proteins that were commonly found in both ITRAQ and SWATH experiments are presented. All raw data generated from the work were submitted and published in the Peptide Atlas public repository (http://www.peptideatlas.org/) for general release (Data ID PASS01507).
机译:阿托品是一种非选择性的毒蕈碱拮抗剂,已知减缓人类青少年和几种动物模型中的近视近视进展。然而,其潜在的分子机制尚不清楚。目前的作品建立了一种单眼形式剥夺近视(FDM)豚鼠模型,使用面部进行FDM眼睛的阿托品治疗2和4周。使用分级的等异常标签的两个时段筛选出对FDM的视网膜蛋白变化以及局部给药的局部施用阿托品的影响,用于与纳米液相色谱 - 串联质谱法(Nano-LC- MS / MS)(n = 24,48只眼)。来自另一种群组的视网膜组织接受4周FDM的与阿托品处理(n = 12,24只眼)进行更显显着的变化,对所有理论质谱(SWATH-MS)蛋白质组学进行顺序窗口,以进行进一步的蛋白质靶标确认。使用ITRAQ和SWATH方法分别鉴定了总共1695个蛋白质(8875肽)和5961蛋白(51871肽)。呈现蛋白质聚合物软件中的Paragon算法,介绍了在ITRAQ和SWATH实验中常见的三种最具明显上调和下调的蛋白质。从工作中生成的所有原始数据都在普通版本(数据ID Pass01507)中提交并发布了肽阿特拉斯公共存储库(http://www.peptidlas.org/)。

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