title: Qualitative and quantitative metabolomic investigation of single neurons by capillary electrophoresis electrospray ionization mass spectrometry.
abstract: Single-cell mass spectrometry (MS) empowers metabolomic investigations by decreasing analytical dimensions to the size of individual cells and subcellular structures. We describe a protocol for investigating and quantifying metabolites in individual isolated neurons using single-cell capillary electrophoresis (CE) coupled to electrospray ionization (ESI) time-of-flight (TOF) MS. The protocol requires ∼2 h for sample preparation, neuron isolation and metabolite extraction, and 1 h for metabolic measurement. We used the approach to detect more than 300 distinct compounds in the mass range of typical metabolites in various individual neurons (25-500 μm in diameter) isolated from the sea slug (Aplysia californica) central and rat (Rattus norvegicus) peripheral nervous systems. We found that a subset of identified compounds was sufficient to reveal metabolic differences among freshly isolated neurons of different types and changes in the metabolite profiles of cultured neurons. The protocol can be applied to the characterization of the metabolome in a variety of smaller cells and/or subcellular domains.
authors.affiliation: 1] Department of Chemistry and the Beckman Institute, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA. [2].
authors.forename: Peter
authors.initials: P
authors.lastname: Nemes
authors.forename: Stanislav S
authors.initials: SS
authors.lastname: Rubakhin
authors.forename: Jordan T
authors.initials: JT
authors.lastname: Aerts
authors.forename: Jonathan V
authors.initials: JV
authors.lastname: Sweedler
mesh_terms: D000818:Animals; D001048:Aplysia / Q000166:cytology; D019075:Electrophoresis, Capillary / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods*; D055442:Metabolome; D055432:Metabolomics / Q000379:methods*; D009474:Neurons / Q000378:metabolism*; D051381:Rats; D059010:Single-Cell Analysis / Q000379:methods*
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