Fig. 1
From: Localized microglia dysregulation impairs central nervous system myelination in development
Digital spatial transcriptomic profiling of human developmental white matter. A Diagram of experimental approach. Regions of interest (ROI) were assessed in infant brain white matter samples (cerebellum), with control ROI (cROI) in control infants compared to ROI in injured infants (iROI) which were normally myelinating (iROI-M+) or showed reduced myelination (iROI-M−). A probe library targeting 1825 genes in 55 pathways was applied to the tissue, and 12 ROIs were selected per sample based on levels of myelin basic protein (MBP; green) and microglia/macrophages (CD68; red), counterstained with Hoechst (cyan). Probes were cleaved with UV light and RNA counts performed per ROI. B Example ROIs from cROI, iROI-M+, and iROI-M− stained for MBP (green) and CD68 (red) and counterstained for Hoechst (blue). Scale bar, 65 µm. C MBP Disruption as measured by average MBP Sphericity per infant in injured iROI-M+ (grey), iROI-M− (pink) and cROI (green). D MBP Disruption as measured by average MBP Sphericity in all infant ROIs, with highest level in control indicated as threshold (dotted line). E Average CD68 count in all infant ROIs, with highest level in control indicated as threshold (dotted line). F CD68 RNA counts versus CD68 protein intensity measurements in respective injured samples. N = 6 infants, n = 71 ROI. Goodness of fit, r2 = 0.8. Slope is significantly non-zero, P < 0.0001. G Principal component analysis (PCA) plot of ROI from control infants (cROI; green) and injured infants (iROI; pink). H PCA plot of cROI (green), iROI-M+(grey), and iROI-M− (pink). I PCA plot of all ROI showing individual injured cases in distinct colours