Fig. 4: Glucose metabolism is necessary for proper mesoderm migration.
From: Selective utilization of glucose metabolism guides mammalian gastrulation
a, Schematic of mesoderm explant isolation for live imaging of migration dynamics. LS, late streak; OB, no bud. b, Control, PD0325901 or YZ9-treated mesoderm explants after 4 h of live imaging. Red asterisks mark daughter cells that divided since imaging started. TCF/LEF:H2B–GFP signal (cyan) was used for nuclear segmentation and tracking. Scale bars, 40 µm. Plots display migration metrics from AIVIA cell-tracking software across two (PD0325901) or three (all other groups) independent experimental replicates. Cell displacement and mean velocity data points represent unique cell tracks. Data are mean ± s.e.m. Dunnett’s multiple comparison test following ordinary one-way ANOVA; P values are shown. All comparisons are to control (control, n = 104 cells for displacement; n = 105 cells for velocity; 2-DG and BrPA, n = 41 cells; PD0325901, n = 85 cells for displacement, n = 87 cells for velocity; YZ9, n = 20 cells for displacement, n = 61 cells for velocity; azaserine, n = 33 cells for displacement, n = 39 cells for velocity). c, Invadapodia assay of mesoderm explants with 2-DG and BrPA, YZ9 and PD0325901 treatments. Yellow arrowheads show examples of individual punctae demonstrating substrate degradation. Scale bars, 100 µm. Data are mean ± s.e.m. Dunnett’s multiple comparison test following ordinary one-way ANOVA; P values are shown. Control, n = 20 fields; 2-DG and BrPA, n = 7 fields; PD0325901, n = 35 fields; YZ9, n = 38 fields; azaserine, n = 26 fields; 3 independent experiments for all groups. d, ECAD expression in mesoderm explants for treatment groups in c (heat map intensity colours used for better visualization). Three independent experiments for all groups. Scale bars, 100 µm. e, KEGG pathway enrichment analysis of differentially expressed genes (DEGs) in PD0325901- and YZ9-treated mesoderm explants showing specifically downregulated terms. Benjamini-adjusted P values from two-sided Fisher’s exact tests are shown. f, Summary illustration of the two distinct waves of glucose metabolic activity that selectively control cellular processes in mouse gastrulation. AVE, anterior visceral endoderm.
