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小鼠脑内空间代谢组学与同位素示踪实验方案。

Protocol for spatial metabolomics and isotope tracing in the mouse brain.

作者信息

Sontising Watit, Yanchik-Slade Francine, Rodriguez-Navas Carlos, Hossen Md Amir, Gonzalez-Rodriguez Manuel, Vaynshteyn Jake, Yamaguchi Shinichi, Boutaghou Mohamed Nazim, Marin-Valencia Isaac

机构信息

The Abimael Laboratory of Neurometabolism, Department of Neurology, Icahn School of Medicine at Mount Sinai, New York, NY, USA.

Shimadzu Scientific Instruments, Columbia, MD, USA.

出版信息

STAR Protoc. 2025 Mar 27;6(2):103716. doi: 10.1016/j.xpro.2025.103716.

DOI:10.1016/j.xpro.2025.103716
PMID:40153438
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11995059/
Abstract

Mass spectrometry imaging enables high-resolution spatial chemical mapping, yet its application for dynamic analysis with tracers poses challenges. Here, we present a protocol for spatial metabolomics and isotope tracing in the mouse brain. We describe steps for tracer administration, tissue collection, and cryosectioning. We then detail procedures for matrix application, ion identification, and data analysis. This protocol delivers high-quality spatial metabolomics data and is well suited for region-specific tracing analysis in the brain.

摘要

质谱成像能够实现高分辨率的空间化学图谱绘制,但其在使用示踪剂进行动态分析方面存在挑战。在此,我们介绍一种用于小鼠大脑空间代谢组学和同位素示踪的方案。我们描述了示踪剂给药、组织采集和冷冻切片的步骤。然后,我们详细说明了基质应用、离子鉴定和数据分析的程序。该方案可提供高质量的空间代谢组学数据,非常适合大脑中特定区域的示踪分析。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d968/11995059/37c5d483ec86/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d968/11995059/bc683ceeeae9/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d968/11995059/79ee0f498bb8/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d968/11995059/6bbca3549502/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d968/11995059/7629ba262885/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d968/11995059/37c5d483ec86/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d968/11995059/bc683ceeeae9/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d968/11995059/79ee0f498bb8/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d968/11995059/6bbca3549502/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d968/11995059/7629ba262885/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d968/11995059/37c5d483ec86/gr4.jpg

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本文引用的文献

1
Imaging brain glucose metabolism in vivo reveals propionate as a major anaplerotic substrate in pyruvate dehydrogenase deficiency.在体成像脑葡萄糖代谢揭示丙酸盐作为丙酮酸脱氢酶缺乏症中的主要补充底物。
Cell Metab. 2024 Jun 4;36(6):1394-1410.e12. doi: 10.1016/j.cmet.2024.05.002.
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Multiscale biochemical mapping of the brain through deep-learning-enhanced high-throughput mass spectrometry.通过深度学习增强的高通量质谱对大脑进行多尺度生化图谱分析。
Nat Methods. 2024 Mar;21(3):521-530. doi: 10.1038/s41592-024-02171-3. Epub 2024 Feb 16.
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RefMet: a reference nomenclature for metabolomics.
RefMet:代谢组学的参考命名法。
Nat Methods. 2020 Dec;17(12):1173-1174. doi: 10.1038/s41592-020-01009-y.
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Brain metabolism modulates neuronal excitability in a mouse model of pyruvate dehydrogenase deficiency.脑代谢在丙酮酸脱氢酶缺乏症小鼠模型中调节神经元兴奋性。
Sci Transl Med. 2019 Feb 20;11(480). doi: 10.1126/scitranslmed.aan0457.
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Correcting for natural isotope abundance and tracer impurity in MS-, MS/MS- and high-resolution-multiple-tracer-data from stable isotope labeling experiments with IsoCorrectoR.使用 IsoCorrectoR 校正 MS、MS/MS 和高分辨率多示踪剂数据中来自稳定同位素标记实验的天然同位素丰度和示踪剂杂质。
Sci Rep. 2018 Dec 17;8(1):17910. doi: 10.1038/s41598-018-36293-4.
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Cardinal: an R package for statistical analysis of mass spectrometry-based imaging experiments.Cardinal:一个用于基于质谱成像实验统计分析的R软件包。
Bioinformatics. 2015 Jul 15;31(14):2418-20. doi: 10.1093/bioinformatics/btv146. Epub 2015 Mar 15.
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The ratio of acetate-to-glucose oxidation in astrocytes from a single 13C NMR spectrum of cerebral cortex.通过大脑皮层的单个碳-13核磁共振波谱得出的星形胶质细胞中乙酸盐与葡萄糖氧化的比率。
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