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Kallabis"],"technology_type":["Mass Spectrometry","Bottom-up proteomics","Data-independent acquisition"],"software":[""],"submitter_keywords":["Synapse","Purinergic receptor","Atp","Tia","Microglia"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD070754"],"tissue":["Cerebrospinal Fluid"],"sample_protocol":["Brain extracellular tissue fluid was collected for 60min using cOFMand immediately frozen on dry ice, before and at 24h, 3dand 7d after TIA.40µl of mouse brain fluid dialysates were prepared according to the single-pot solid-phase-enhanced sample preparation (SP3) protocol(Hughes, Moggridge et al., 2019) with some adaptations. In brief, samples were mixed 1:1 with lysis buffer containing 10% SDS in PBS pH 8.5, 10 mMTris(2-carboxyethyl) phosphine, and 30mM 2-chloroacetamide. Proteins were denatured, reduced, and alkylated by incubating samples at 70°C, 800rpm on a shaker for 10 min.  We added 2µl of prepared magnetic bead mixtures (Sera-Mag Magnetic carboxylate modified particles, Cytiva) to each sample and mixed them 1:1 with 100% acetonitrile (ACN). Samples were incubated for 8min at RT and 800rpm on a shaker. Following, samples were transferred on a magnet and magnetic beads with bound proteins were washed two times with 200µl 70% ethanol and once with 100% ACN while keeping the samples the whole time on the magnet. For enzymatic digestion, the magnetic beads were resuspended in 10µl digestion buffer containing 0.1µg trypsin / Lys-C mixture (Promega) in 50mM ammonium bicarbonate buffer pH 8.0 and digestion at 37°C, 400rpm overnight. On the next day, samples were mixed with 380µl 100% ACN and incubated for 8min at room temperature. After transfer on a magnet, beads were washed with 200µl 100% ACN and beads air-dried. Peptides were eluted from magnetic beads by resuspending the same in 9µl of 5% ACN in LC-MS-grade water. We determined peptide concentrations and used 250ng peptides per sample for each LC-MS run. LC-MS/MS analysis Samples were measured by liquid chromatography-tandem mass spectrometry using an Easy nLC 1200 chromatographic system (Thermo Fisher Scientific) and the Exploris 480 mass spectrometer (Thermo Fisher Scientific). Peptides were separated by 90min chromatographic gradients using a binary buffer system with buffer A (0.1 % formic acid in LC-MS-grade water) and buffer B (80% ACN, 0.1% formic acid in LC-MS-grade water). We used an in-house packed analytical column with a length of 50cm and filled with 1.9µm ReproSil-Pur 120 C18-AQ material (Dr. Maisch). To separate peptides the amount of buffer B was linearly increased from 4% to 25% over 70min with a constant flow rate of 300nl/min. Following, buffer B was increased to 55% over 8min and a sharp increase to 95% buffer B over 2min. The analytical column was washed at 95% for 10min. Eluting peptides were ionized by nano-electrospray ionization at a constant spray voltage of 2.5kV. We measured the samples in data-independent acquisition (DIA) mode. In brief, full MS were recorded at a resolution of 60,000 with an AGC target of 300% and a maximum injection time of 55ms. The scan range was set to 340 – 1,080m/z. DIA MS/MS scans were recorded at a resolution of 15,000, an AGC target of 1,000%, and a maximum injection time of 22ms. The DIA window m/z range was set from 400 – 1,000m/z separated into 50 isolation windows with a size of 12m/z per window. We used a staggered window approach with isolation windows shifted by 6m/z every second scan cycle. Fragment ions were generated with an HCD collision energy of 27%. We measured pooled samples for the generation of a gas phase fractioned (GPF) spectral library. Similar LC-MS methods were applied, only the DIA scan ranges were limited to 100m/z covering the overall m/z range from 400 – 1,000m/z in six consecutive runs and staggered DIA windows with a size of 4m/z."],"repository":["Pride"],"quantification_method":[""],"modification":[""],"data_protocol":["Staggered windows were deconvoluted with the MSConvert tool of the ProteoWizard software suit (v. 3.0.21321, https://www.nature.com/articles/nbt.2377). Spectral library generation and peptide identification/quantification from LC-MS raw data was performed with the DIA-NN software suit (v. 1.8 https://doi.org/10.1038/s41592-019-0638-x). We used the SWISS-PROT Mus musculus fasta database downloaded from UniProt (v. 2021-11-18) to make a spectral library using the six GPF measurements from pooled samples. Trypsin was set as the digestion enzyme with a maximum of one miss-cleavage and cysteine carbamidomethylation was set as a fixed modification. The scan window radius was set to 10, mass accuracies were fixed to 2e-05 (MS2) and 7.5e-06 (MS1), respectively. Precursor peptides were filtered at an FDR <1%. Label-free normalization of protein groups was performed in R using the MaxLFQ algorithm(Cox, Hein et al., 2014) and proteotypic peptides only.  Statistical analysis of the data was performed with the Perseus software suite (v. 1.6.15,(Tyanova, Temu et al., 2016)). In brief, we log2 transformed protein LFQ intensities and filtered proteins for data completeness in at least one time point. Missing values were replaced sample-wise by random drawing of numbers from 1.8 standard deviations downshifted, and 0.3 standard deviations broad normal distributions. Quantile normalization was performed with an R Perseus-plugin as well as a batch correction with ComBat R Perseus-plugin (batch = animal). Significantly regulated proteins were identified by ANOVA multiple sample testing (S0 = 0.1, permutation-based FDR = 0.05, 250 randomizations). ANOVA-significant proteins were further Z-score normalized and used for hierarchical clustering using Euclidean distances. We searched systematically for enriched processes in identified clusters using Fisher‘s exact testing (Benjamini-Hochberg FDR = 0.02)."],"omics_type":["Proteomics"],"labhead":["Arthur Liesz"],"instrument_platform":[""],"submission_type":["PARTIAL"],"labhead_affiliation":["Institute for Stroke and Dementia Research, LMU Feodor-Lynen-Str. 17, 81377 Munich, Germany"],"species":["Mus Musculus (mouse)"],"publication":["10.1038/S44321-026-00397-6"],"submitter_mail":["kallabis@uni-bonn.de"],"submitter_affiliation":["Core Facility Translational Proteomics\nMedical Faculty\nUniversity Bonn\nVenusberg - Campus 1\n53127 Bonn Germany"],"submitter_country":["Germany"],"additional_accession":[]},"is_claimable":false,"name":"Blocking microglial reactivity via purinergic receptors prevents subacute cognitive deficits after TIA","description":"Our research presents a new animal model of transient ischemic attack (TIA) that mimics brief episodes without cell loss, but results in neuronal and behavioral deficits. We identified excessive microglial reactivity, driven by acute ATP release, as a key factor in post-TIA neurological deficits, which were ameliorated by inhibiting the P2Y12 receptor, a microglia-specific purinergic receptor in the brain parenchyma responsible for activity-dependent microglial cell-cell interactions. This finding suggests that modulation of microglial reactivity offers a promising strategy to prevent cognitive impairment in TIA patients, opening avenues for future research in this underexplored area.","dates":{"publication":"2026-04-24","submission":"2025-11-14"},"accession":"PXD070754","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:377960","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:1073882","NEWT:475174","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:1590","NEWT:10034","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:376619","NCBITaxon:79857","NEWT:1096976","NEWT:3765","NEWT:1589","NEWT:135622","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656064","NEWT:36630","NEWT:383379","NEWT:418106","NCBITaxon:12721","NEWT:10029","NEWT:913645","NEWT:641809","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:7719","NEWT:868565","NEWT:135674","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:108061","NEWT:60711","NEWT:224308","NEWT:3347","NEWT:160621","NEWT:212790","NEWT:110368","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:44688","NEWT:44689","NEWT:498211","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:44685","NEWT:161934","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:410661","NEWT:1140","NCBITaxon:2157","NEWT:1143","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:2065263","NEWT:177437","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:244366","NEWT:1246791","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:33548","NEWT:52299","NEWT:8165","NEWT:96794","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:55784","NEWT:1303443","NEWT:485","NEWT:56636","NEWT:2853422","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:1502","NEWT:128017","NEWT:376686","NEWT:95486","NEWT:1883446","NEWT:1233435","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:491","NEWT:460523","NEWT:235443","NEWT:1757312","NEWT:85694","NEWT:108458","NEWT:5936","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:1116234","NEWT:1255228","NEWT:410289","NEWT:373153","NEWT:472","NEWT:940825","NEWT:529507","NEWT:1071661","NEWT:470","NEWT:5911","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:445","NEWT:153009","NEWT:261756","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:215402","NEWT:1547","NEWT:906914","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:150475","NEWT:267872","NEWT:381666","NEWT:172269","NEWT:9534","NEWT:5180","NEWT:256737","NEWT:9541","NEWT:8694","NEWT:33936","NEWT:8692","NEWT:2903","NEWT:185579","NEWT:13076","NEWT:1006581","NEWT:8673","NEWT:33940","NEWT:550","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:36185","NEWT:1054211","NEWT:1225786","NEWT:51338","NEWT:575412","NEWT:28112","NEWT:6493","NEWT:6494","NEWT:6491","NEWT:507601","NEWT:520","NEWT:186441","NEWT:643680","NEWT:214092","NCBITaxon:6157","NEWT:13095","NEWT:162425","NEWT:104105","NEWT:216257","NEWT:9986","NEWT:8654","NEWT:8658","NEWT:1268063","NEWT:8657","NEWT:9983","NEWT:8655","NEWT:5147","NEWT:28108","NEWT:28104","NEWT:407821","NEWT:568703","NCBITaxon:2","NEWT:568708","NEWT:986","NEWT:52641","NEWT:28532","NEWT:353152","NEWT:298176","NEWT:227377","NEWT:1774284","NEWT:40674","NEWT:1194669","NEWT:443906","NEWT:519","NEWT:2510939","NEWT:7394","NEWT:6063","NEWT:1328388","NEWT:105487","NEWT:1548728","NEWT:667127","NEWT:9557","NEWT:377586","NEWT:300641","NEWT:39655","NEWT:9554","NEWT:475932","NEWT:256318","NEWT:206411","NCBITaxon:6191","NEWT:229533","NEWT:214053","NEWT:80863","NEWT:90675","NEWT:52638","NEWT:57075","NEWT:1666905","NEWT:8697","NEWT:8695","NEWT:884204","NEWT:1123869","NEWT:9544","NEWT:9545","NEWT:979","NEWT:7370","NEWT:83906","NEWT:99488","NEWT:1134506","NEWT:255470","NEWT:38783","NEWT:6426","NEWT:33090","NEWT:9935","NEWT:287889","NEWT:305959","NEWT:13489","NEWT:29829","NEWT:92867","NEWT:92866","NCBITaxon:3055","NEWT:51750","NEWT:202950","NEWT:48106","NEWT:295027","NCBITaxon:11320","NEWT:632957","NEWT:9925","NCBITaxon:9606","NEWT:90690","NEWT:1436183","NEWT:300273","NEWT:4232","NEWT:416348","NEWT:11298","NEWT:4236","NEWT:7742","NEWT:196627","NEWT:200308","NEWT:200302","NEWT:870435","NEWT:9913","NEWT:9915","NEWT:105841","NEWT:2666255","NEWT:999810","NCBITaxon:5693","NEWT:28995","NEWT:1392998","NEWT:380394","NEW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