A microanalytical capillary electrophoresis mass spectrometry assay for quantifying angiotensin peptides in the brain
A microanalytical capillary electrophoresis mass spectrometry assay for quantifying angiotensin peptides in the brain
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DOI：
10.1007/s00216-019-01771-9
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发表时间：
2019-07-01
影响因子：
4.3
通讯作者：
Nemes, Peter
中科院分区：
文献类型：
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作者：
Lombard-Banek, Camille;Yu, Zhe;Nemes, Peter
大脑的肾素-血管紧张素系统（RAS）产生一系列生物活性的血管紧张素来源的肽，参与生理稳态和疾病的病理生理。尽管迄今为止有大量的研究工作，但对大脑RAS生理学的全面了解尚不充分。一个重大的挑战是，有限的生物分析分析方法能够在生理低浓度（2-15fmol/g湿组织）下检测血管紧张素（Ang）肽，并具有足够的化学特异性来进行明确的分子鉴定。此外，复杂的大脑解剖结构要求对特定组织区域进行微观分析，从而进一步提高了RAS研究中识别和量化的灵敏度要求。为了填补这一技术空白，我们开发了一种微量分析方法，将实验室建立的毛细管电泳（CE）纳米电喷雾电离（nano-ESI）平台与高分辨率质谱仪（HRMS）相结合。使用平行反应监测，我们证明了该技术实现了自信的鉴定和定量的Ang肽在大约。灵敏度为5ammol至300zmol。这种微量分析方法揭示了从下丘脑皮层下器官和室旁核（参与口渴和水分平衡以及神经内分泌调节压力的重要大脑区域）中微采样的组织之间的Ang肽谱差异。微分析ce -纳米esi - hrms扩展了神经科学的分析工具箱，以帮助更好地了解RAS。
The renin-angiotensin system (RAS) of the brain produces a series of biologically active angiotensinogen-derived peptides involved in physiological homeostasis and pathophysiology of disease. Despite significant research efforts to date, a comprehensive understanding of brain RAS physiology is lacking. A significant challenge has been the limited set of bioanalytical assays capable of detecting angiotensin (Ang) peptides at physiologically low concentrations (2-15fmol/g of wet tissue) and sufficient chemical specificity for unambiguous molecular identifications. Additionally, a complex brain anatomy calls for microanalysis of specific tissue regions, thus further taxing sensitivity requirements for identification and quantification in studies of the RAS. To fill this technology gap, we here developed a microanalytical assay by coupling a laboratory-built capillary electrophoresis (CE) nano-electrospray ionization (nano-ESI) platform to a high-resolution mass spectrometer (HRMS). Using parallel reaction monitoring, we demonstrated that this technology achieved confident identification and quantification of the Ang peptides at approx. 5amol to 300 zmol sensitivity. This microanalytical assay revealed differential Ang peptide profiles between tissues that were micro-sampled from the subfornical organ and the paraventricular nucleus of the hypothalamus, important brain regions involved in thirst and water homeostasis and neuroendocrine regulation to stress. Microanalytical CE-nano-ESI-HRMS extends the analytical toolbox of neuroscience to help better understand the RAS.