Fig. 2: Tanycyte module expression maps to the spatial distribution lining the third ventricle.
a UMAP clustering of tanycytes (β2-, β1-, α1-, α2-Tanycytes) from the Day 5 single-cell RNA-sequencing (scRNA-seq) dataset. Cell type annotations were inferred from a published dataset16. b Tanycyte lineage cells plotted on principal components 1 and 2 revealed a trajectory (left). The colors represent the subtype of each cell from a. A principal curve fit across the trajectory was used to assign a pseudoventricle score to each cell corresponding to the spatial location of that cell type along third ventricular (3V) wall. Density plot of each tanycyte subpopulation plotted along the pseudoventricle revealed a continuum of gene expression and non-distinct shifts in the expression of marker genes along the pseudoventricle. c Scaled module expression of sc-tany-M1 in FGF1-treated (black) and vehicle (gray) tanycytes (linear mixed-effects model; bin size of 1; mean ± SEM; *p < 0.05) (Bonferroni adjusted) from the Day 5 scRNA-seq dataset (n = 6 mice/group). Exact adjusted p values are provided in Supplementary Data 5. ME module eigengene. d Gene–gene network plot of the correlation of top 15 genes in module sc-tany-M1 (left) and associated pathways (right). Genes mentioned in the text are labeled in red. e Representative images of immunolabeling of vimentin in diabetic Lepob/ob mice 1 week after vehicle or icv FGF1 injection. Pixel dimensions: 0.27 × 0.27 × 2.00 microns. Quantification: dots represent volume of vimentin-positive voxels assessed by iMARIS for individual animals (mean from 2 sections per animal, n = 4/group), mean ± SEM. Unpaired t test, two tailed, *p = 0.022 (not adjusted). Source data are provided as a Source data file. f Scaled module expression of sc-tany-M2, sc-tany-M3, sc-tany-M5, sc-tany-M6, and sc-tany-M23 with 5 consecutive points of difference in expression along the pseudoventricle between FGF1-treated (black) and vehicle (gray) tanycytes (linear mixed-effects models; bin size of 1; mean ± SEM; *p < 0.05) (Bonferroni adjusted) from the Day 5 scRNA-seq dataset (n = 6 mice/group). Exact adjusted p values are provided in Supplementary Data 5.
