Journal: EMBO Molecular Medicine

Article Title: Blocking microglial reactivity via purinergic receptors prevents subacute cognitive deficits after TIA

doi: 10.1038/s44321-026-00397-6

Figure Lengend Snippet: ( A ) Representative scheme of the microperfusion system. ( B ) Heat map of all ANOVA+ proteins of the LC-MS analysis of the perfusate products, ( C ) protein intensity over time, top ten upregulated genes and pathway analysis from cluster 3 and ( D ) cluster 5 in males ( n = 5 per group). ( E ) Uniform manifold approximation and projection (UMAP) plot, colored by identified clusters (different cell types). ( F ) Number of differentially expressed genes ( p < 0.05) at 24 h and 3 d after TIA in each cell type compared to control (naïve) in males ( n = 3 per group). ( G ) Volcano plot of differentially regulated genes in microglia 3 d compared to naïve animals. Cut-off for illustrating gene names: Log2FC >|2| illustrated by vertical gray dashed lines, adjusted p value: >−log10(0.05) illustrated by horizontal gray dashed line ( n = 3 per group). ( H ) Representative image and quantification of microglia (P2Y12R + ) and neuron soma (KV2.1 + ) contacts 24 h after TIA in males ( n = 5 per group; p value = 0.0079), scale bar = 20 µm. ( I ) Representative image of microglia (P2Y12 + ) and synapse (Vglut + Homer + ) staining for synapse engulfed quantification 24 h after TIA in males ( n = 5 per group; p value = 0.0079), scale bar = 1 µm. Statistical tests: ( B – D ) Two-way ANOVA, Fisher’s exact testing. ( G ) Wilcoxon rank sum test + Bonferroni correction. ( H , I ) Two-way Student’s t -test. Error bars indicate ±SD. * p < 0.05. .

Article Snippet: For the microglia-synapse analysis, confocal Z-stack images were processed, and colocalizations of synapses (VGlut+ Homer+) in contact with microglial cells (P2Y12+) were analyzed using Imaris software.

Techniques: Liquid Chromatography with Mass Spectroscopy, Control, Staining

Journal: EMBO Molecular Medicine

Article Title: Blocking microglial reactivity via purinergic receptors prevents subacute cognitive deficits after TIA

doi: 10.1038/s44321-026-00397-6

Figure Lengend Snippet: ( A ) Luciferace assay to detect ATP concentration from peri-lesional perfusate (see Fig. ) at before (BL) and at indicated time points after TIA in males ( n = 6 per group; 10 min: p value = 0.0002, 24 h: p value = 0.0231). ( B ) Quantification of extracellular ATP (hsyn-cATPs) using in vivo two-photon imaging before (BL) and at indicated time points after TIA ( n = 18 per group; 10 min: p value <0.01) and ( C ) microglial processes (Cx3Cr1cre-TdTomato) recruitment to focal increase in extracellular ATP within 3 min after TIA in males. ( D ) Representative image of Iba1+ microglia cell and 3D reconstructed microglia for branches nodes analysis 24 h after cisterna magna injection of different ATP concentrations in males ( n = 6 per group; 0.3, 1, and 3 µg ATP: p value <0.0001), scale bar = 20 µm. ( E ) Quantification of colocalized presynaptic and postsynaptic particles 24 h after cisterna magna injection of different ATP concentrations in males ( n = 6 per group; 0.3, 1, and 3 µg ATP: p value <0.0001). ( F ) Microglia sphericity ( n = 5 per group; Veh: p value <0.0001 and P2Y12Ri: p value <0.0001), ( G ) synapse (Vglut + Homer + ) contacts with microglial cells (P2Y12 + ) ( n = 10 per group; Veh: p value <0.0033 and P2Y12Ri: p value <0.0035) (scale bar = 2 µm) and ( H ) glutamatergic synapse counts ( n = 5 per group) in sham and 24 h after TIA in vehicle (Veh) or P2Y12Ri (-inhibitor) treated animals (in males’ mice); Veh: p value = 0.0140 and P2Y12Ri: p value = 0.0155. ( I ) Neuroscore at 48 h after TIA in vehicle (Veh) or P2Y12Ri (-inhibitor) treated animals ( n = 10 per group; p value = 0.0002) and ( J ) Barnes Maze test ( n = 10 per group) in Sham and 48 h after TIA in vehicle (Veh) or P2Y12Ri (-inhibitor) treated animals (in males’ mice); Veh: p value = 0.0083 and P2Y12Ri: p value = 0.0469. Statistical tests: ( A – J ) two-way ANOVA, corrected for multiple comparisons using two-stage step-up method of Benjamin Kriegel. (I) Mann–Whitney U -test. Error bars indicate ±SD. * p < 0.05. .

Article Snippet: For the microglia-synapse analysis, confocal Z-stack images were processed, and colocalizations of synapses (VGlut+ Homer+) in contact with microglial cells (P2Y12+) were analyzed using Imaris software.

Techniques: Concentration Assay, In Vivo, Imaging, Injection, MANN-WHITNEY