Jorg Hanrieder 1, Grzegorz Wicher 2, Jonas Bergquist 1, Asa Fex Svennigsen 3, Malin Andersson 4
1Department of Physical and Analytical Chemistry, Analytical Chemistry, Uppsala University, Uppsala, Sweden
2Department of Neuroscience, Developmental Genetics, Uppsala University, Uppsala, Sweden
3Institute of Medical Biology, Anatomy and Neurobiology, University of Southern Denmark, Odense, Denmark
4Department of Pharmaceutical Bioscience, Toxicology, Uppsala University, Uppsala, Sweden |
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Development of new microanalytical strategies for molecular characterization, particularly of CNS cells is of central relevance in current neuroscience research. Molecular characterization enables monitoring of cell growth stages, neuronal stem cell differentiation as well as probing cellular responses towards different kinds of stimulation.
Matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI TOF MS) exhibits excellent features allowing sensitive, robust and specific analysis of large biomolecules in complex matrices.
Here we describe a fast and robust methodology for direct characterization of intact mammalian glial cells and visualization in developing mammalian brain using MALDI TOF MS. Glial cells, were placed on the MALDI target and analyzed directly resulting in characteristic mass fingerprint spectra that allowed significant distinction of different cell types and facilitated identification of cell specific protein mass peaks. These identifier masses allowed in turn to visualize oligodendrocytes localization in brain tissue with molecular specificity using imaging mass spectrometry. The observed protein identities were further confirmed by bottom up proteomic profiling. Validation of protein identities, protein regulation and localization in nervous tissue was performed by immunochemistry. |
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