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Single-Cell Mass Spectrometry of Metabolites and Proteins for Systems and Functional Biology. 系统和功能生物学中代谢物和蛋白质的单细胞质谱分析。
Neuromethods
Pub Date : 2022-01-01 DOI: 10.1007/978-1-0716-2525-5_5
Erika P Portero, Leena Pade, Jie Li, Sam B Choi, Peter Nemes

Molecular composition is intricately intertwined with cellular function, and elucidation of this relationship is essential for understanding life processes and developing next-generational therapeutics. Technological innovations in capillary electrophoresis (CE) and liquid chromatography (LC) mass spectrometry (MS) provide previously unavailable insights into cellular biochemistry by allowing for the unbiased detection and quantification of molecules with high specificity. This chapter presents our validated protocols integrating ultrasensitive MS with classical tools of cell, developmental, and neurobiology to assess the biological function of important biomolecules. We use CE and LC MS to measure hundreds of metabolites and thousands of proteins in single cells or limited populations of tissues in chordate embryos and mammalian neurons, revealing molecular heterogeneity between identified cells. By pairing microinjection and optical microscopy, we demonstrate cell lineage tracing and testing the roles the dysregulated molecules play in the formation and maintenance of cell heterogeneity and tissue specification in frog embryos (Xenopus laevis). Electrophysiology extends our workflows to characterizing neuronal activity in sections of mammalian brain tissues. The information obtained from these studies mutually strengthen chemistry and biology and highlight the importance of interdisciplinary research to advance basic knowledge and translational applications forward.

分子组成与细胞功能错综复杂地交织在一起，阐明这种关系对于理解生命过程和开发下一代治疗方法至关重要。毛细管电泳(CE)和液相色谱(LC)质谱(MS)的技术创新通过允许高特异性的无偏检测和分子定量，提供了以前无法获得的细胞生物化学见解。本章介绍了我们经过验证的方案，将超灵敏质谱与细胞、发育和神经生物学的经典工具相结合，以评估重要生物分子的生物学功能。我们使用CE和LC MS测量脊索动物胚胎和哺乳动物神经元中单个细胞或有限群体组织中的数百种代谢物和数千种蛋白质，揭示鉴定细胞之间的分子异质性。通过显微注射和光学显微镜的结合，我们展示了细胞谱系追踪，并测试了失调分子在青蛙胚胎(Xenopus laevis)细胞异质性和组织规格的形成和维持中的作用。电生理学将我们的工作流程扩展到表征哺乳动物脑组织部分的神经元活动。从这些研究中获得的信息相互加强了化学和生物学，并突出了跨学科研究对推进基础知识和转化应用的重要性。
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引用次数: 0
Magnetic Resonance Imaging as a Translational Research Tool for Major Depression 磁共振成像作为重性抑郁症的转化研究工具
Neuromethods
Pub Date : 2022-01-01 DOI: 10.1007/978-1-0716-2083-0_12
Chien-Han Lai
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引用次数: 0
Molecular Tools to Study Regeneration of the Avian Cochlea and Utricle 研究鸟类耳蜗和耳蜗再生的分子工具
Neuromethods
Pub Date : 2022-01-01 DOI: 10.1007/978-1-0716-2022-9_5
Amanda Janesick, Mirko Scheibinger, S. Heller
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引用次数: 1
Imaging Neuronal Activity in Cerebellar Cortex of Behaving 行为的小脑皮层神经元活动成像
Neuromethods
Pub Date : 2022-01-01 DOI: 10.1007/978-1-0716-2026-7_13
M. Kislin, G. Broussard, B. Deverett, S. Wang
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引用次数: 0
Translational Strategies for Developing Biomarkers for Major Depression: Lessons Learned from Animal Models 开发重度抑郁症生物标志物的转化策略:从动物模型中获得的经验教训
Neuromethods
Pub Date : 2022-01-01 DOI: 10.1007/978-1-0716-2083-0_5
F. Aricioglu, B. Leonard
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引用次数: 0
Electrophysiological Recordings of Voltage-Dependent and Mechanosensitive Currents in Sensory Hair Cells of the Auditory and Vestibular Organs of the Mouse 小鼠听觉和前庭器官感觉毛细胞中电压依赖性和机械敏感电流的电生理记录
Neuromethods
Pub Date : 2022-01-01 DOI: 10.1007/978-1-0716-2022-9_10
Artur A. Indzhykulian, Stuart L. Johnson, G. Géléoc
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引用次数: 1
An Efficient Method to Detect Messenger RNA (mRNA) in the Inner Ear by RNAscope In Situ Hybridization 一种利用RNAscope原位杂交检测内耳信使RNA (mRNA)的有效方法
Neuromethods
Pub Date : 2022-01-01 DOI: 10.1007/978-1-0716-2022-9_6
Sumana Ghosh, Graham R. Casey, Kendra L. Stansak, P. Thapa, Bradley J Walters
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引用次数: 1
Measuring Cerebellar Processing and Sensorimotor Functions in Non-Human 非人类小脑加工和感觉运动功能的测量
Neuromethods
Pub Date : 2022-01-01 DOI: 10.1007/978-1-0716-2026-7_14
Nico A. Flierman, Eric Avila, C. D. De Zeeuw, A. Badura
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引用次数: 0
Neurobiochemistry Alterations Associated with Major Depression: A Review of Translational Magnetic Resonance Spectroscopic Studies 与重度抑郁症相关的神经生化改变:磁共振波谱研究综述
Neuromethods
Pub Date : 2022-01-01 DOI: 10.1007/978-1-0716-2083-0_13
D. Roddy, John R. Kelly, Thomas Drago, Kesidha Raajakesary, Madeline Haines, E. O'Hanlon
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引用次数: 2
Behavioral Paradigms for Assessing Cognitive Functions in the Chronic Social Defeat Stress Model of Depression 评估抑郁症慢性社会挫折应激模型认知功能的行为范式
Neuromethods
Pub Date : 2022-01-01 DOI: 10.1007/978-1-0716-2083-0_7
M. Fakhoury, Michael Fritz, Sama F. Sleiman
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引用次数: 5
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