Fig. 3: Origins of astrocyte diversity from regional and scRNA-seq data.

(A) Experimental procedure for scRNA-seq. Single cells were dissociated from the cortex, hippocampus, and striatum of wild type mice (N = 4-7 mice). Data of striatal cells were partly from our recent study (34) and combined with new cortical, hippocampal, and striatal single cell data. (B) Uniform manifold approximation and projection (UMAP) plot of 89,553 brain cells from 3 CNS regions grouped by expression similarity identified 26 major cell populations (10 major cell types). (C) UMAP plot of cluster analysis for 8,898 astrocytes (AST1 and AST2 in panel B) from 3 CNS regions. The cell numbers of astrocyte subclusters are shown as AST1 to AST7. (D) UMAP plot of the astrocyte subclusters to illustrate the three different brain regions examined. (E) UMAP plots from panel C showing expression of Chrdl1, Gfap, and Crym. (F). The heat map shows the top 100 genes that were upregulated in each astrocyte subcluster relative to the others. Example genes are identified. (G) Proportional bar graph showing the percentage of each astrocyte subcluster representation in the 3 CNS regions from scRNA-seq data. (H) Correlations between astrocyte-region specific RNA-seq data (left), bulk RNA-seq data (middle), and bulk RNA-seq minus astrocyte region specific RNA-seq (right) that were gathered as part of Fig 2. The correlation plots show that astrocytes grouped into three broad anatomical areas (a, b, and c).