GPMDB00204OtherBell-Temin H, et al.InstrumentNot AvailableNot availableMus_musculus_viruses, Mousesmstevens@usf.edu26424600http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32320019111http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32320019132http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32320019104http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32320019125http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32320019146http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32320019139University of South FloridaNot availableGPMDBMicroglia, the resident immune cells of the brain, have been shown to display a complex spectrum of roles that span from neurotrophic to neurotoxic depending on their activation status. Microglia can be classified into four stages of activation, M1, which most closely matches the classical (pro-inflammatory) activation stage, and the alternative activation stages M2a, M2b, and M2c. The alternative activation stages have not yet been comprehensively analyzed through unbiased, global-scale protein expression profiling. In this study, BV2 mouse immortalized microglial cells were stimulated with agonists specific for each of the four stages and total protein expression for 4644 protein groups was quantified using SILAC-based proteomic analysis. After validating induction of the various stages through a targeted cytokine assay and Western blotting of activation states, the data revealed novel insights into the similarities and differences between the various states. The data identify several protein groups whose expression in the anti-inflammatory, pro-healing activation states are altered presumably to curtail inflammatory activation through differential protein expression, in the M2a state including CD74, LYN, SQST1, TLR2, and CD14. The differential expression of these proteins promotes healing, limits phagocytosis, and limits activation of reactive nitrogen species through toll-like receptor cascades. The M2c state appears to center around the down-regulation of a key member in the formation of actin-rich phagosomes, SLP-76. In addition, the proteomic data identified a novel activation marker, DAB2, which is involved in clathrin-mediated endocytosis and is significantly different between M2a and either M1 or M2b states. Western blot analysis of mouse primary microglia stimulated with the various agonists of the classical and alternative activation states revealed a similar trend of DAB2 expression compared with BV2 cells.Novel Molecular Insights into Classical and Alternative Activation States of Microglia as Revealed by Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC)-based Proteomics.Bell-Temin Harris H,Culver-Cochran Ashley E AE,Chaput Dale D,Carlson Christina M CM,Kuehl Melanie M,Burkhardt Brant R BR,Bickford Paula C PC,Liu Bin B,Stevens Stanley M SM Jr,Bell-Temin Harris H,Culver-Cochran Ashley E AE,Chaput Dale D,Carlson Christina M CM,Kuehl Melanie M,Burkhardt Brant R BR,Bickford Paula C PC,Liu Bin B,Stevens Stanley M SM,Bell-Temin Harris H, Culver-Cochran Ashley E AE, Chaput Dale D, Carlson Christina M CM, Kuehl Melanie M, Burkhardt Brant R BR, Bickford Paula C PC, Liu Bin B, Stevens Stanley M SMMF.microglia.CNS, mixed glial cells (predominantly microglia), microglial cell, brain macrophage, hortega cells, Hortega cell, brain-resident macrophage., activation, microgliocyte, microgliathe brain, 3.4.22.-, scale tissue, PLXN5, Entire brain, mol, Laboratory, developmental stage, peltate hair, Mus domesticus, Nl1, PLEXIN-B1, mini-ICE, CASP-14, Encephalons, PPS1, House Mouse, SeP, hToll, CEH, House, Mus musculus domesticus, NEPII, NL1, synganglion, NL2, SEH, Mice, SEP, MF.microglia.CNS, study, Ly87, Brains, Hortega cell, cell, Swiss, Lps, OFC6, LPS, plant peltate hair, PPS, Swiss Mice, proteins, suprasegmental levels of nervous system, "mouse" EXACT common_name [], sEP, "Mus muscaris" RELATED misnomer [], CD284, lipopolysaccharides, SELP, "mice C57BL/6xCBA/CaJ hybrid" RELATED misspelling [], stage, house mouse, PIT, mixed glial cells (predominantly microglia), data, suprasegmental structures, brain-resident macrophage, mouse, Mell1, VWS1, encephalon, Cell, mice C57BL/6xCBA/CaJ hybrid, Mus muscaris, polypeptide, Experiment, MMEL2, Mus, Mini-ICE, proteomic analysis, VWS, scales, activation, TLR4, Rasl2-8, Mus musculus, microglial cell, scale, Caspase-14 subunit p10, mice, Encephalon, proteomic analysis., Swiss Mouse, House Mice, Caspase-14 subunit p19, Ran|M1, MICE, microgliocyte, microglia, Laboratory Mice, domesticus, "house mouse" EXACT genbank_common_name [], Eph2, brain macrophage, hortega cells, Mouse, Laboratory Mouse, NEP2MF.microglia.CNS, mixed glial cells (predominantly microglia), microglial cell, amino acids, Hortega cell, Aminokarbonsaeure, brain-resident macrophage, Aminocarbonsaeure, Isotope Labeling, Amino, Isotope-Coded Affinity, Affinity Tagging, microgliocyte, Isotopically-Coded, Labeling, microglia, Aminosaeure., Isotope, Stable, Stable Isotope, Isotope-Coded Affinity Tagging, Isotope-Coded, brain macrophage, hortega cells, Acids, Isotopically-Coded Affinity, Tagging, activation, Stable Isotope Labeling, Isotope Coded Affinity Tagging, Isotopically-Coded Affinity Taggingthe brain, 3.4.22.-, CIB8, scale tissue, AA407514, Entire brain, Laboratory, MHC class II-associated invariant chain, Ia antigen-associated invariant chain, developmental stage, peltate hair, Mus domesticus, clathrin-mediated endocytosis, Receptors, Gene, mini-ICE, CASP-14, biosynthesis, Encephalons, HLADG, CME, House Mouse, Downregulation, Monocyte differentiation antigen CD14, II, House, urinary form, fam:TLR, Gene Products, Mus musculus domesticus, monocyte differentiation antigen CD14, Cell., synganglion, JTK8, B8, Toll Like, Mice, Doc2, DOC2, Reactive, multicellular organismal biosynthetic process, Ii, MF.microglia.CNS, study, Reactive Nitrogen, Toll Like Receptor, single-organism biosynthetic process, Phagolysosomes, Brains, Hortega cell, formation, Phagosome, twm, Swiss, anabolism, plant peltate hair, CLIP, Ly105, Doc-2, Phagolysosome, Swiss Mice, proteins, myeloid cell-specific leucine-rich glycoprotein, suprasegmental levels of nervous system, D15Wsu122e, "mouse" EXACT common_name [], study assay, CD14, Toll Like Receptors, DOC-2, RCB0750, synthesis, "Mus muscaris" RELATED misnomer [], CD282, scientific observation, Hck-2, "mice C57BL/6xCBA/CaJ hybrid" RELATED misspelling [], D630005B22Rik, stage, house mouse, clathrin-dependent endocytosis, mixed glial cells (predominantly microglia), measuring, HLA-DR antigens-associated invariant chain, clathrin coated pit-dependent endocytosis, SLP76, data, suprasegmental structures, p96, DHLAG, brain-resident macrophage, Proteins, mouse, p56Lyn, Toll-Like Receptor, encephalon, Cell, Species, mice C57BL/6xCBA/CaJ hybrid, Down Regulation, Mus muscaris, polypeptide, any method, SLP-76, Down-Regulation, Receptor Down-Regulation, AA960054, Mus, Myeloid cell-specific leucine-rich glycoprotein, Down-Regulation (Physiology), Mini-ICE, Protein, proteomic analysis, membrane-bound form, p53Lyn, scales, TIL4, Phagocytoses, activation, 5730435J12Rik, TLR2, Mus musculus, microglial cell, Toll-Like, scale, Caspase-14 subunit p10, mice, Nitrogen Species, Encephalon, Swiss Mouse, p33, House Mice, Caspase-14 subunit p19, MICE, Toll/interleukin-1 receptor-like protein 4, AI323664, microgliocyte, BB161688, microglia, Laboratory Mice, domesticus, Protein Gene Products, "house mouse" EXACT genbank_common_name [], CD74, Gene Proteins, AI957090, Ia-GAMMA, brain macrophage, hortega cells, Mouse, Receptor, Laboratory Mouse2004http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32320019104dbgap_ncbi~0patentfamilies~0rfam~0merops~0complex-portal~0uniprot~0wormbaseparasite~0embl-covid19~0reactome~0emdb~0wgs_masters~0ebiweb_resources~0opentargets_genetics~0biomodels_all~0ipd-mhc~0ebiweb_teams~0taxonomy~0genome_assembly~0sc-experiments~0ebiweb_people~0enzymeportal_enzymes~0ipd-nhkir~0cellosaurus~0pdbe~0chebi~0patentproteins~0interpro7~0uniref~0chembl~0pdbekb~0gpcrdb~0hgnc~0sc-genes~0intact~0rhea~0ebiweb_training~0alphafold~0imgt-hla~0patentnucleotides~0ensemblroot~0eva_studies~0non-coding~0europepmc~0pubmed~1identifiers_registry~0pdbechem~0hpa-covid19~0eva-variants-covid19~0biosamples~0gwas_catalog~0biotools~0tls_masters~0mesh~0coding~0sra~0opentargets~0efo~0embl-pathogen~0project~0human_diseases~0geo_datasets~0embl~0treefam~0uniparc~0ols~0dgva~0intenz~0go~0tsa_masters~0biosamples-covid19~0ebiweb_corporate~0omim~0lrg~0earlycause-molecular-sequences~0ipd-kir~0empiar~0rnacentral~0orcid_data_claims~0gpmdb~3lineage-covid19~0metagenomics~0pfam~0varsite~00.00.00.0061690314620604570.001.0falseNovel molecular insights into classical and alternative activation states of microglia as revealed by SILAC-based proteomics.Data from ProteomeXchange, 888 Proteomic analysis of microglial activation states, 1676 Microglial activation characterization. Experiment: BV2_LPS_1, file: folder summary. Published as part of Mol Cell Proteomics. 2015 Sep 30 . From the Abstract: {{i}} Microglia, the resident immune cells of the brain, have been shown to display a complex spectrum of roles that span from neurotrophic to neurotoxic depending on their activation status. Microglia can be classified into four stages of activation, M1, which most closely matches the classical (pro-inflammatory) activation stage, and the alternative activation stages M2a, M2b, and M2c. The alternative activation stages have not yet been comprehensively analyzed through unbiased, global-scale protein expression profiling. In this study, BV2 mouse immortalized microglial cells were stimulated with agonists specific for each of the four stages and total protein expression for 4,644 protein groups was quantified using SILAC-based proteomic analysis ... {{/i}}2015-10-08GPM3232001910426424600