Journal: Nature Communications
Article Title: Molecular determinants of nephron vascular specialization in the kidney
Figure Lengend Snippet: Analysis of vascular heterogeneity development. a Pseudotime trajectory of vascular differentiation in the kidney. VP, vascular progenitor; PTC, peritubular capillary; AA/LA, afferent arteriole/large arteries (pre-glomerular); AVR/V, ascending vasa recta/venous blood vessels; EA, efferent arteriole; GC, glomerular capillaries; DVR, descending vasa recta. b Heat maps denoting genes that become differentially expressed as pre-glomerular arteries branch from vascular progenitor cells. Notable genes of venous peritubular progenitor capillaries ( Cryab , Igfbp5 , Plvap ) and arteries ( Jag1 , Fbln5 , Gja5 ) are shown on the right. c Heat maps denoting genes that become differentially expressed as glomerular capillaries (GC) and postglomerular arteries branch from embryonic progenitor capillaries which mature into peritubular capillaries. Notable genes of glomerular capillaries ( Sema5a , Lpl ), postglomerular arteries ( Aqp1 , Slc14a1 ), and peritubular capillaries ( Igfbp5 , Plvap ) are on the right. d – g Pseudo-time trajectory plots denoting the expression of genes enriched in particular vascular clusters. Plots include Aplnr in vascular progenitor cells (VP) ( d ), Plvap in veins (V), VPs, and peritubular capillaries (PTC)( e ), Sox17 in afferent arterioles/large pre-glomerular arteries (AA/LA) and descending vasa recta/efferent arterioles (DVR/EA)( f ), and Lpl in glomerular capillaries (GC)( g ). h Immunofluorescent staining of the apelin receptor ( Aplnr ) in E17 kidney. Endothelial cells were stained with endomucin ( Emcn ). SSB, s-shaped body; VP, vascular progenitor; Angio., angiogenic vessel; GC, glomerular capillary; V, vein; A, arteriole. Scale bars: first panel 40 μm, second panel 5 μm, third and fourth panel 10 μm. i Pie chart denoting mean fluorescent intensity of Aplnr antibody staining in peritibular capillaries, veins, arteries, and glomerular capillaries. n = 3, average of 5 frames of view. j R26R-Confetti E18 mouse kidneys cut in half sagittally after tamoxifen induction at E11. GC, glomerular capillary; PTC, peritubular capillary. Scale bars: first panel 100 μm, second to sixth panel 5 μm. k Bar graph denoting the number of fluorescent reporters found in identified vascular structures. n > 10 for each structure. Art, Arteries; VR, Vasa Recta. l E13 mouse kidney showing the primary vascular plexus exists as generic capillaries (the vascular progenitor cells) before subvascular specification at E14-E15 stages. Dotted lines outline the cortex and medulla. Endothelial cells are stained with endomucin ( Emcn ) and the outer cortex of the kidney is denoted by Six2 staining of nephron progenitors. Scale bar 100 μm.
Article Snippet: The following antibodies were used: chicken anti-GFP (for Flk-GFP; Aves, GFP-1020, 1:500), goat anti-Cx40/Gja5 (Santa Cruz, sc-20466, 1:100), rabbit anti-Collagen IV (Millipore, AB756P, 1:400), goat anti-Nrp1 (R & D Systems, AF566, 1:100), rat anti-Plvap (BD Pharmingen, 550563, 1:100), goat anti-Sox17 (R & D Systems, AF1924, 1:100), rat anti-Endomucin (Santa Cruz, sc-65495, 1:100), rabbit anti-Aquaporin1 (Biorad, MCA2100, 1:100), rabbit anti-Tbx3 (Abcam, ab99302, 1:100), rabbit anti-Aplnr (Protein Tech, 20341–1-AP, 1:100), goat anti-Igfbp5 (R & D Systems, AF578, 1:100), goat anti-Igfbp7 (Abcam, ab129302, 1:100), rabbit anti-Six2 (Protein Tech, 11562–1-AP, 1:100).
Techniques: Expressing, Staining