Metabolic Profiling and Quantification of Neurotransmitters in Mouse Brain by Gas Chromatography-Mass Spectrometry

Q1 Agricultural and Biological Sciences

Current protocols in mouse biology Pub Date : 2016-09-01 DOI:10.1002/cpmo.15

Christian Jäger, Karsten Hiller, Manuel Buttini

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引用次数: 10

Abstract

Metabolites are key mediators of cellular functions, and have emerged as important modulators in a variety of diseases. Recent developments in translational biomedicine have highlighted the importance of not looking at just one disease marker or disease inducing molecule, but at populations thereof to gain a global understanding of cellular function in health and disease. The goal of metabolomics is the systematic identification and quantification of metabolite populations. One of the most pressing issues of our times is the understanding of normal and diseased nervous tissue functions. To ensure high quality data, proper sample processing is crucial. Here, we present a method for the extraction of metabolites from brain tissue, their subsequent preparation for non-targeted gas chromatography-mass spectrometry (GC-MS) measurement, as well as giving some guidelines for processing of raw data. In addition, we present a sensitive screening method for neurotransmitters based on GC-MS in selected ion monitoring mode. The precise multi-analyte detection and quantification of amino acid and monoamine neurotransmitters can be used for further studies such as metabolic modeling. Our protocol can be applied to shed light on nervous tissue function in health, as well as neurodegenerative disease mechanisms and the effect of experimental therapeutics at the metabolic level. © 2016 by John Wiley & Sons, Inc.

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用气相色谱-质谱法测定小鼠脑内神经递质的代谢谱和定量

代谢物是细胞功能的关键介质，已成为多种疾病的重要调节剂。转化生物医学的最新发展突出了不只是关注一种疾病标志物或疾病诱导分子，而是关注其群体的重要性，以便全面了解细胞在健康和疾病中的功能。代谢组学的目标是系统地鉴定和量化代谢物群体。我们这个时代最紧迫的问题之一是了解正常和患病的神经组织功能。为了确保高质量的数据，适当的样品处理是至关重要的。在这里，我们提出了一种从脑组织中提取代谢物的方法，随后将其用于非靶向气相色谱-质谱(GC-MS)测量，并给出了一些原始数据处理的指导方针。此外，我们提出了一种在选择离子监测模式下基于GC-MS的神经递质敏感筛选方法。氨基酸和单胺类神经递质多分析物的精确检测和定量可用于代谢模型等进一步研究。我们的方案可以应用于阐明健康的神经组织功能，以及神经退行性疾病的机制和实验治疗在代谢水平的影响。©2016 by John Wiley &儿子,Inc。

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来源期刊

Current protocols in mouse biology Agricultural and Biological Sciences-Animal Science and Zoology

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期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet, all too often, nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. The aim of Current Protocols in Mouse Biology is to provide the clearest, most detailed and reliable step-by-step instructions for protocols involving the use of mice in biomedical research. Written by experts in the field and extensively edited to our exacting standards, the protocols include all of the information necessary to complete an experiment in the laboratory—introduction, materials lists with supplier information, detailed step-by-step procedures with helpful annotations, recipes for reagents and solutions, illustrative figures and information-packed tables. Each article also provides invaluable discussions of background information, applications of the methods, important assumptions, key parameters, time considerations, and tips to help avoid common pitfalls and troubleshoot experiments. Furthermore, Current Protocols in Mouse Biology content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Mouse Biology brings together resources in mouse biology and genetics and provides a mouse protocol resource that covers all aspects of mouse biology. Current Protocols in Mouse Biology also permits optimization of mouse model usage, which is significantly impacted by both cost and ethical constraints. Optimal and standardized mouse protocols ultimately reduce experimental variability and reduce the number of animals used in mouse experiments.