title: Single-neuron identification of chemical constituents, physiological changes, and metabolism using mass spectrometry.
abstract: The use of single-cell assays has emerged as a cutting-edge technique during the past decade. Although single-cell mass spectrometry (MS) has recently achieved remarkable results, deep biological insights have not yet been obtained, probably because of various technical issues, including the unavoidable use of matrices, the inability to maintain cell viability, low throughput because of sample pretreatment, and the lack of recordings of cell physiological activities from the same cell. In this study, we describe a patch clamp/MS-based platform that enables the sensitive, rapid, and in situ chemical profiling of single living neurons. This approach integrates modified patch clamp technique and modified MS measurements to directly collect and detect nanoliter-scale samples from the cytoplasm of single neurons in mice brain slices. Abundant possible cytoplasmic constituents were detected in a single neuron at a relatively fast rate, and over 50 metabolites were identified in this study. The advantages of direct, rapid, and in situ sampling and analysis enabled us to measure the biological activities of the cytoplasmic constituents in a single neuron, including comparing neuron types by cytoplasmic chemical constituents; observing changes in constituent concentrations as the physiological conditions, such as age, vary; and identifying the metabolic pathways of small molecules.
authors.affiliation: CAS Key Laboratory of Urban Pollutant Conversion, School of Chemistry and Materials Science, University of Science and Technology of China, Hefei, Anhui 230026, China.|School of Life Sciences, University of Science and Technology of China, Hefei 230026, China.
authors.forename: Hongying
authors.initials: H
authors.lastname: Zhu
authors.affiliation: School of Life Sciences, University of Science and Technology of China, Hefei 230026, China.
authors.forename: Guichang
authors.initials: G
authors.lastname: Zou
authors.affiliation: School of Life Sciences, University of Science and Technology of China, Hefei 230026, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Wang
authors.affiliation: CAS Key Laboratory of Urban Pollutant Conversion, School of Chemistry and Materials Science, University of Science and Technology of China, Hefei, Anhui 230026, China.
authors.forename: Meihui
authors.initials: M
authors.lastname: Zhuang
authors.affiliation: School of Life Sciences, University of Science and Technology of China, Hefei 230026, China; wxiong@ustc.edu.cn gmhuang@ustc.edu.cn.|Neurodegenerative Disorder Research Center, University of Science and Technology of China, Hefei 230026, China.|CAS Key Laboratory of Brain Function and Disease and Hefei National Laboratory for Physical Sciences at the Microscale, University of Science and Technology of China, Hefei 230026, China.|Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Xiong
authors.affiliation: CAS Key Laboratory of Urban Pollutant Conversion, School of Chemistry and Materials Science, University of Science and Technology of China, Hefei, Anhui 230026, China; wxiong@ustc.edu.cn gmhuang@ustc.edu.cn.
authors.forename: Guangming
authors.initials: G
authors.lastname: Huang
mesh_terms: D000818:Animals; D003593:Cytoplasm / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods*; D053858:Metabolic Networks and Pathways / Q000235:genetics; D009474:Neurons / Q000737:chemistry*; D018408:Patch-Clamp Techniques / Q000379:methods; D059010:Single-Cell Analysis / Q000379:methods*
keywords: mass spectrometry; metabolism; patch clamp; single neuron