Journal: Cell Reports Medicine
Article Title: Single-cell spatial transcriptomics reveals a dynamic control of metabolic zonation and liver regeneration by endothelial cell Wnt2 and Wnt9b
doi: 10.1016/j.xcrm.2022.100754
Figure Lengend Snippet: Single-cell spatial transcriptomics profiling by Molecular Cartography reveals dynamic zonation changes in EC-Wnt2-9b-DKO mice (A) UMAP plots of single-cell transcriptomic analysis of Molecular Cartography data showing genetic clusters of hepatocytes from control and EC-Wnt2-9b-DKO livers based on expression of 16 zonated genes. Seurat clustering identifies C1-C6 clusters with C1 and C2 clusters representing hepatocytes expressing pericentral (zone 3) genes, C3 representing hepatocytes expressing midzonal genes, and C4-C6 representing hepatocytes expressing periportal genes. An enrichment of C4-C6 hepatocytes is observed in the EC-Wnt2-9b-DKO cluster at the expense of C1 and C2 clusters. (B) UMAP plots using spatial location of cells in Molecular Cartography analysis based on nine arbitrary but equivalent segments manually drawn between portal vein and central vein shows various clusters recognized in (A) to represent zonal location validating the accuracy of this technique in addressing single-cell spatial transcriptomics. (C) Molecular Cartography comparing spatial gene expression of Glul , Lect2 , Axin2 , Cldn2 , Cyp1a2 , Oat , Cyp2e1 , and Rgn (zone 3), and Fbp1 , G6pc , Pck1 , Gls2 , Arg1 , Ass1 , Cps1 , and Cyp2f2 (zone 1), and Sox9 (mostly in cholangiocytes) in control and EC-Wnt2-9b-DKO liver showing loss of zone 3 genes and overall “periportalization” of the DKO liver. CV, central vein; PV, portal vein. (D) Line plots showing impaired pericentral expression of Wnt target genes (shown here are Glul , Oat , Cyp2e1 , and Axin2) in DKOs compared with control. PC, pericentral; Mid, midzonal; PP, periportal. (E) Line plots showing dynamic changes in the expression of periportal genes (shown here are Fads1 , Gls2 , Cps1 , Elovl2 , and Ass1) in DKOs compared with control. Red arrow, upregulation; green arrow, downregulation; negative, no change. Color stands for the extent of up- or downregulation. (F) Molecular Cartography depicting altered spatial expression of Ccnd1 and Igfbp2 in EC-Wnt2-9b-DKO versus control liver. While the expression of both genes is midzonal with some overlap, both genes are located pericentrally in DKOs with Ccnd1 being more restricted to hepatocytes next to the central vein. CV, central vein; PV, portal vein. (G) Line plots of three representative midzonal genes ( Ccnd1 , Pon1 , and Igfbp2 ) showing altered location to pericentral hepatocytes in DKOs compared with control. The hue around line plots represents SEM.
Article Snippet: Rabbit monoclonal anti-Cyclin D1 , Abcam , Cat#ab134175; RRID: AB_2750906.
Techniques: Expressing