Journal: Communications Biology
Article Title: Maternal immune activation followed by peripubertal stress combinedly produce reactive microglia and confine cerebellar cognition
doi: 10.1038/s42003-025-07566-2
Figure Lengend Snippet: a Schematic drawing of the imaging mass cytometry (IMC). Polymer-conjugated antibodies (Methods) are combined with labeling metals, which visualize the sharp spectrum by time of flight. b A multiplexed IMC image of the four-μm-thick cerebellar cortex. c Antibody-marker list. d IMC images obtained from cerebellar cortex (culmen and simple lobule), dentate nucleus (DN), and fastigial nucleus (FN)+medial vestibular nuclei (MVN) with IBA1, IL6ST/gp130, MMP9, TREM2, claudin-5, and MHC-class II markers. Yellow squares in 2HIT show representative 2HIT stress-associated puncta (arrowheads, Methods). Cerebellar cortex (Cbl Crtx). Cerebellar nuclei (Cbl Nuc). e , f An increase in MHC-class II(+) positive immune cells in the 2HIT cerebellum. The immune cell density was bar-graphed in different regions of Control and 2HIT. g–i An increase in stress-associated puncta in the cerebellum. The number of puncta was counted in two cerebellar regions (cerebellar cortex and dentate nucleus) of 2HIT. Density (# per millimeter square) and the histograms binned by diameter are shown. The total area of the cerebellar cortex ( g ) and dentate nucleus ( h ) are 3.609 and 1.235 mm 2 , respectively. MHC-class II(+) microglia located near or colocalized to Claudin-5(+) vasculature ( i ). The histogram of density vs. distance to the capillary is shown at the bottom of ( i ).
Article Snippet: For IMC with a human patient’s post-mortem brain (Supplementary Fig. ), we used the following antibodies for disease-associated microglia/macrophages (DAMs) markers (Mouse anti-Iba1 Monoclonal antibody (Invitrogen, MA5-38265, clone: HL22-MS, RRID: AB_2898181), Rabbit anti-gp130/IL6ST Polyclonal antibody (Sino Biological, 101867-T10, RRID: AB_3094604), Mouse anti-TREM2 Monoclonal antibody (NSJ Bio, V9417SAF, clone: TREM2/7210, RRID: AB_3094605), Mouse anti-Apolipoprotein E (APOE) Monoclonal antibody (abcam, ab1906, clone: D6E10, RRID: AB_302668), and mouse anti-MHC class II Monoclonal antibody (abcam, ab55152, clone: 6C6, AB_944199)).
Techniques: Imaging, Mass Cytometry, Polymer, Labeling, Marker, Control
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