实验性胼胝体脱髓鞘和髓鞘再生过程中的蛋白质组学变化。
Proteomic changes during experimental de- and remyelination in the corpus callosum.
机构信息
Department of Biochemistry and Medical Chemistry, University of Pécs Medical School, Pécs, Hungary.
Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark.
出版信息
PLoS One. 2020 Apr 9;15(4):e0230249. doi: 10.1371/journal.pone.0230249. eCollection 2020.
BACKGROUND
In the cuprizone model of multiple sclerosis, de- and remyelination can be studied without major interference from the adaptive immune responses. Since previous proteomic studies did not focus on the corpus callosum, where cuprizone causes the most pronounced demyelination, we performed a bottom up proteomic analysis on this brain region.
METHODS
Eight week-old mice treated with 0.2% cuprizone, for 4 weeks and controls (C) were sacrificed after termination of the treatment (4wD), and 2 (2dR) or 14 (2wR) days later. Homogenates of dissected corpus callosum were analysed by quantitative proteomics. For data processing, clustering, gene ontology analysis, and regulatory network prediction, we used Perseus, PANTHER and Ingenuity Pathway Analysis softwares, respectively.
RESULTS
We identified 4886 unmodified, single- or multi phosphorylated and/or gycosylated (PTM) proteins. Out of them, 191 proteins were differentially regulated in at least one experimental group. We found 57 proteins specific for demyelination, 27 for early- and 57 for late remyelinationwhile 36 proteins were affected in two, and 23 proteins in all three groups. Phosphorylation represented 92% of the post translational modifications among differentially regulated modified (PTM) proteins with decreased level, while it was only 30% of the PTM proteins with increased level. Gene ontology analysis could not classify the demyelination specific proteins into any biological process category, while allocated the remyelination specific ones to nervous system development and myelination as the most specific subcategory. We also identified a protein network in experimental remyelination, and the gene orthologues of the network were differentially expressed in remyelinating multiple sclerosis brain lesions consistent with an early remyelination pattern.
CONCLUSION
Proteomic analysis seems more informative for remyelination than demyelination in the cuprizone model.
背景
在多发性硬化症的铜诱导模型中，可以在不受适应性免疫反应主要干扰的情况下研究脱髓鞘和髓鞘再生。由于之前的蛋白质组学研究并未集中在胼胝体上，而铜诱导模型在该区域造成最明显的脱髓鞘，因此我们对该脑区进行了下向蛋白质组学分析。
方法
8 周龄的小鼠用 0.2%铜诱导剂处理 4 周，然后分为对照组（C）和实验组（4 周停药组，4wD），停药后 2 天（2dR）或 14 天（2wR）处死。对分离的胼胝体匀浆进行定量蛋白质组学分析。使用 Perseus、PANTHER 和 Ingenuity Pathway Analysis 软件分别用于数据处理、聚类、基因本体分析和调控网络预测。
结果
我们鉴定出 4886 个未经修饰的、单或多磷酸化和/或糖基化的（PTM）蛋白。其中，有 191 种蛋白在至少一个实验组中发生差异调控。我们发现 57 种蛋白特异性地参与脱髓鞘，27 种蛋白参与早期髓鞘再生，57 种蛋白参与晚期髓鞘再生，而 36 种蛋白在两组中均受影响，23 种蛋白在三组中均受影响。在差异调节的修饰（PTM）蛋白中，磷酸化占差异调节的修饰（PTM）蛋白水平降低的 92%，而在水平升高的 PTM 蛋白中仅占 30%。基因本体分析不能将脱髓鞘特异性蛋白分类到任何生物学过程类别中，而将髓鞘再生特异性蛋白分配到神经系统发育和髓鞘化作为最特异的亚类别。我们还鉴定了实验性髓鞘再生中的蛋白质网络，并且网络的基因直系同源物在髓鞘再生多发性硬化症脑病变中表达差异，与早期髓鞘再生模式一致。
结论
蛋白质组学分析在铜诱导模型中对髓鞘再生的信息比脱髓鞘更丰富。
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