Optically Guided Single Cell Mass Spectrometry of Rat Dorsal Root Ganglia to Profile Lipids, Peptides and Proteins

Thanh D. Do, Joseph F. Ellis, Elizabeth K. Neumann, Troy J. Comi, Emily G. Tillmaand, Ashley E. Lenhart, Stanislav S. Rubakhin, Jonathan V. Sweedler

Research output: Contribution to journalArticlepeer-review

Abstract

The mammalian dorsal root ganglia (DRG) are located on the dorsal roots of the spinal nerves and contain cell bodies of primary sensory neurons. DRG cells have been classified into subpopulations based on their size, morphology, intracellular markers, response to stimuli, and neuropeptides. To understand the connections between DRG chemical heterogeneity and cellular function, we performed optically guided, high-throughput single cell profiling using sequential matrix-assisted laser desorption/ionization mass spectrometry (MS) to detect lipids, peptides, and several proteins in individual DRG cells. Statistical analysis of the resulting mass spectra allows stratification of the DRG population according to cellular morphology and, presumably, major cell types. A subpopulation of small cells contained myelin proteins, which are abundant in Schwann cells, and mass spectra of several larger cells contained peaks matching neurofilament, vimentin, myelin basic protein S, and thymosin beta proteins. Of the over 1000 cells analyzed, approximately 78 % produced putative peptide-rich spectra, allowing the population to be classified into three distinct cell types. Two signals with m/z 4404 and 5487 were exclusively observed in a cell type, but could not be matched to results of our previous liquid chromatography-MS analyses.

Original languageEnglish (US)
Pages (from-to)1180-1191
Number of pages12
JournalChemPhysChem
Volume19
Issue number10
DOIs
StatePublished - May 22 2018

Keywords

  • dorsal root ganglia
  • mass spectrometry
  • neurochemistry
  • peptides
  • single cell analysis

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics
  • Physical and Theoretical Chemistry

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