Fig. 7: Injury induces GLAST-expressing pericyte and perivascular fibroblast activation and the generation of myofibroblasts.
a, UMAP plot of tdTomato+EGFP+ cells isolated from the uninjured and injured spinal cord of GLAST-CreERT2;R26R-tdTomato;Pdgfrb-eGFP mice at 3 and 5 days after injury grouped by injury state (left) and cell population (right). b, Violin plots showing the expression levels of cell population-specific genes. c, Left, feature plots showing the expression of the fibroblast marker Pi16 and pericyte marker Atp13a5. Right, pseudotime trajectory analysis of tdTomato+EGFP+ cells isolated from the uninjured and injured spinal cord at 3 and 5 days after injury. Dashed lines outline cell populations. High expression of Pi16 and Atp13a5 were used to identify pseudotime starting points in the fibroblast and pericyte populations, respectively. d,e, Highlight of the pseudotime trajectory of the fibroblast branch (d) and pericyte branch (e). f, Gene expression profile of Acta2, Col5a1 and Postn along pseudotime within the fibroblast branch. g, Gene expression profile of Atp13a5, Cspg4 and Col1a1 along pseudotime within the pericyte branch. h, Left, violin plots showing the gene expression levels of Atp13a5 and Col1a1 per cell population and coexpression graph of Col1a1 and Atp13a5. Right, violin plots showing the gene expression levels of Kcjn8 and Col5a1 per cell population and coexpression graph of Col5a1 and Kcnj8. Each dot in coexpression graphs represents a single cell, colored according to the injury state. i,j, GO analyses based on upregulated genes of activated pericytes and myofibroblasts (i) and upregulated genes of myofibroblasts and activated fibroblasts (j). Venn diagrams represent the overlap of common GO terms and unique GO terms between the indicated populations. Relevant GO terms per comparison are indicated below. k, Dot plot of upregulated marker genes per fibroblast population. d3, day 3; d5, day 5; Per, pericyte; aPer, activated pericyte; pFib, perivascular fibroblast; mFib, myofibroblast; aFib, activated fibroblast; NA, nonattributed; TNF, tumor necrosis factor; MHC, major histocompatibility complex; IGF, insulin-like growth factor; IGFBP, insulin-like growth factor-binding protein; SLC, solute carrier; HIF-1, hypoxia-inducible factor. Source data are provided as a source data file.
