Extracellular Alterations in pH and K+ Modify the Murine Brain Endothelial Cell Total and Phospho-Proteome.
Extracellular Alterations in pH and K+ Modify the Murine Brain Endothelial Cell Total and Phospho-Proteome.
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DOI：
10.3390/pharmaceutics14071469
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发表时间：
2022-07-15
期刊：
影响因子：
5.4
通讯作者：
Largent-Milnes, Tally M.
中科院分区：
文献类型：
--
作者：
Wahl, Jared R.;Vivek, Anjali;Palomino, Seph M.;Almuslim, Moyad;Cottier, Karissa E.;Langlais, Paul R.;Streicher, John M.;Vanderah, Todd W.;Liktor-Busa, Erika;Largent-Milnes, Tally M.
血脑屏障（BBB）的病理与许多中枢神经系统（CNS）疾病有关，而这些疾病的病理尚不清楚。皮质扩张性抑制（CSD）长期以来被认为与这些疾病状态的潜在机制有关，但尚未完全理解。本研究旨在利用脑内皮细胞（b.End3）小鼠内皮瘤细胞的血脑屏障（BBB）体外模型，通过表征主要前感觉物质释放到中枢神经系统细胞外空间的影响，研究CSD在血脑屏障病理中的作用。应用跨内皮电阻（TEER）筛选、跨细胞摄取和免疫反应方法，结合整体蛋白质组学和磷酸化蛋白质组学方法，评估体外模拟CSD事件对模拟血脑屏障的影响。研究结果表明与肌动蛋白细胞骨架和内皮紧密连接相关的蛋白质的重新定位和功能改变。此外，在磷酸化蛋白质组学分析中发现，CSD过程中释放的单个物质诱导的独特病理机制具有独特的磷酸化特征，这表明Zona Occludins 1 （ZO-1）可能是血脑屏障的病理“检查点”。利用这些磷酸化特征，可能会开发出新的CSD诊断方法，值得进一步研究。
Pathologies of the blood–brain barrier (BBB) have been linked to a multitude of central nervous system (CNS) disorders whose pathology is poorly understood. Cortical spreading depression (CSD) has long been postulated to be involved in the underlying mechanisms of these disease states, yet a complete understanding remains elusive. This study seeks to utilize an in vitro model of the blood–brain barrier (BBB) with brain endothelial cell (b.End3) murine endothelioma cells to investigate the role of CSD in BBB pathology by characterizing effects of the release of major pronociceptive substances into the extracellular space of the CNS. The application of trans-endothelial electrical resistance (TEER) screening, transcellular uptake, and immunoreactive methods were used in concert with global proteome and phospho-proteomic approaches to assess the effect of modeled CSD events on the modeled BBB in vitro. The findings demonstrate relocalization and functional alteration to proteins associated with the actin cytoskeleton and endothelial tight junctions. Additionally, unique pathologic mechanisms induced by individual substances released during CSD were found to have unique phosphorylation signatures in phospho-proteome analysis, identifying Zona Occludins 1 (ZO-1) as a possible pathologic “checkpoint” of the BBB. By utilizing these phosphorylation signatures, possible novel diagnostic methods may be developed for CSD and warrants further investigation.
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影响因子：
6.3
作者：
Andrew, R. David;Hsieh, Yi-Ting;Brisson, C. Devin
通讯作者：
Brisson, C. Devin
影响因子：
14.5
作者：
Dreier, Jens P.;Woitzik, Johannes;Strong, Anthony J.
通讯作者：
Strong, Anthony J.
影响因子：
8
作者：
Blasig, IE;Winkler, L;Piontek, J
通讯作者：
Piontek, J
影响因子：
2.9
作者：
Aanismaa, Paivi;Gatlik-Landwojtowicz, Ewa;Seelig, Anna
通讯作者：
Seelig, Anna
影响因子：
7
作者：
Kruse, Rikke;Krantz, James;Langlais, Paul R.
通讯作者：
Langlais, Paul R.