Analytical and Bioanalytical Chemistry

, Volume 401, Issue 1, pp 135–147

MALDI mass spectrometry based molecular phenotyping of CNS glial cells for prediction in mammalian brain tissue

Original Paper

DOI: 10.1007/s00216-011-5043-y

Cite this article as:
Hanrieder, J., Wicher, G., Bergquist, J. et al. Anal Bioanal Chem (2011) 401: 135. doi:10.1007/s00216-011-5043-y

Abstract

The development of powerful analytical techniques for specific molecular characterization of neural cell types is of central relevance in neuroscience research for elucidating cellular functions in the central nervous system (CNS). This study examines the use of differential protein expression profiling of mammalian neural cells using direct analysis by means of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). MALDI-MS analysis is rapid, sensitive, robust, and specific for large biomolecules in complex matrices. Here, we describe a newly developed and straightforward methodology for direct characterization of rodent CNS glial cells using MALDI-MS-based intact cell mass spectrometry (ICMS). This molecular phenotyping approach enables monitoring of cell growth stages, (stem) cell differentiation, as well as probing cellular responses towards different stimulations. Glial cells were separated into pure astroglial, microglial, and oligodendroglial cell cultures. The intact cell suspensions were then analyzed directly by MALDI-TOF-MS, resulting in characteristic mass spectra profiles that discriminated glial cell types using principal component analysis. Complementary proteomic experiments revealed the identity of these signature proteins that were predominantly expressed in the different glial cell types, including histone H4 for oligodendrocytes and S100-A10 for astrocytes. MALDI imaging MS was performed, and signature masses were employed as molecular tracers for prediction of oligodendroglial and astroglial localization in brain tissue. The different cell type specific protein distributions in tissue were validated using immunohistochemistry. ICMS of intact neuroglia is a simple and straightforward approach for characterization and discrimination of different cell types with molecular specificity.

Keywords

Intact cell mass spectrometry (ICMS)MALDI-TOF-MSImaging mass spectrometry (IMS)Glial cells

Supplementary material

216_2011_5043_MOESM1_ESM.pdf (307 kb)
ESM 1(PDF 306 kb)

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.Department of Pharmaceutical Bioscience, Drug Safety and Toxicology and Department of Physical and Analytical Chemistry, Analytical ChemistryUppsala UniversityUppsalaSweden
  2. 2.Department of Genetics and Pathology, Cancer and Vascular BiologyUppsala UniversityUppsalaSweden
  3. 3.Department of Physical and Analytical Chemistry, Analytical ChemistryUppsala UniversityUppsalaSweden
  4. 4.Department of Pharmaceutical Bioscience, Drug Safety and ToxicologyUppsala UniversityUppsalaSweden
  5. 5.Institute of Medical Biology, Neurobiology ResearchUniversity of Southern DenmarkOdenseDenmark