Aktivitet: Deltagelse i faglig begivenhed › Organisering af eller deltagelse i konference
MALDI TOF MS profiling of central nervous system glia
G. WICHER, J. HANRIEDER, J. BERGQUIST, M. ANDERSSON, *A. FEX SVENNINGSEN
Development of specific microanalytical strategies for molecular cell type characterization is of central relevance in current neuroscience research. Molecular phenotyping enables analysis of cell developmental stages as well as probing cellular responses to different stimulations. Matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI TOF MS) provide excellent profiling capabilities allowing rapid, sensitive, robust and specific analysis of large biomolecules in complex matrixes. Here we report a fast and robust approach for direct characterization of intact central nervous system glial cells and prediction of cellular phenotypes in rodent brain tissue using MALDI TOF MS. Astrocytes, oligodendrocytes and microglia cell cultures were analyzed directly resulting in specific mass profile-spectra that allowed significant distinction of different cell types and facilitated identification of cell specific protein mass peaks. These identifier masses allowed in turn prediction of oligodendroglia localization in brain tissue with molecular specificity. MALDI MS profiling analysis revealed a 4-5-fold higher histone peak intensity/cell in oligodendrocytes compared with astrocytes and even higher compared with microglia, including histone H2A, H2B, H3, and H4. Furthermore, mass peaks of 11308 Da, and 11350 Da were found to appear only in oligodendrocyte spectra. These masses are suggested to match the histone H4 (2 x methylated and singly acetylated: 11308 and doubly acetylated: 11350 respectively). We have verified MALDI MS results performing immuno-labeling against histone variants on primary cultures and tissue from rat brain. This "proof of concept'' study, examines the use of differential protein expression profiling of the different types of rodent CNS glia that enable profiling of these glial cells in injury or disease.