Qualitative and quantitative metabolomic investigation of single neurons by capillary electrophoresis electrospray ionization mass spectrometry

• Published in: Nature protocols Vol. 8; no. 4; pp. 783 - 799
• Main Authors: Nemes, Peter, Rubakhin, Stanislav S, Aerts, Jordan T, Sweedler, Jonathan V
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01-04-2013; Nature Publishing Group
• Subjects: 631/1647/2230/1452; 631/1647/320; 631/1647/527/296; 631/378; Analytical Chemistry; Animals; Aplysia - cytology; Biological Techniques; Capillary electrophoresis; Cells; Chemical properties; Computational Biology/Bioinformatics; Electrophoresis; Electrophoresis, Capillary - methods; Ionization; Lasers; Life Sciences; Mass spectrometry; Mass Spectrometry - methods; Metabolism; Metabolites; Metabolome; Metabolomics; Metabolomics - methods; Methods; Microarrays; Neurons; Neurons - metabolism; Organic Chemistry; Physiology; Protocol; Quantitative chemical analysis; Rats; Sample preparation; Scientific imaging; Single-Cell Analysis - methods; Time-of-flight mass spectrometry; United States; United States > US; Illinois
• ISSN: 1754-2189; 1750-2799; 1750-2799
• DOI: 10.1038/nprot.2013.035

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.