Journal: Human Molecular Genetics

Article Title: The planar cell polarity gene Vangl2 is required for mammalian kidney-branching morphogenesis and glomerular maturation

doi: 10.1093/hmg/ddq397

Figure Lengend Snippet: Early branching morphogenesis is disrupted in Vangl2 Lp kidneys in vivo and in vitro . Haematoxylin- and eosin-stained transverse sections through whole E11.5 embryos showed similar appearances in both wild-type ( A ) and Vangl2 Lp/Lp ( B ) littermate embryos ( n = 3 each genotype). Both had paired metanephric rudiments in which the ureteric bud (UB) penetrated into metanephric mesenchyme (MM). E13.5 whole z-stack images of wild-type ( C ) and Vangl2 Lp/Lp ( D ) kidneys immunostained for cytokeratin (green signal). Inserts in (C) and (D) show representative zoomed-in images of typical ureteric bud tips; Vangl2 Lp/Lp bud tips are frequently shorter and broader than in wild-type. ( E ) There was a significant ( P < 0.01) reduction in numbers of ureteric bud branch tips in Vangl2 Lp heterozygote ( n = 12) and homozygote ( n = 5) versus wild-type ( n = 4) metanephroi. At E18.5, Vangl2 Lp kidneys ( F, right) appeared smaller and abnormally shaped in comparison to wild-type littermates (F, left). After 2days in culture, E11.5 mutant explants ( H and I ) appeared variably growth-retarded versus wild-type organs ( G ). Furthermore, these cultured Vangl2 Lp/Lp metanephroi ( n = 10) had significantly ( P = 0.009) fewer bud tips in comparison to wild-type littermates ( n = 7) ( J ). ( K and L) To assess potential further differentiation in vitro , E12.0 metanephroi were cultured for 5 days and then immunostained for both cytokeratin (red) to highlight the arborizing ureteric tree, and WT1 (green) to highlight podocytes within glomeruli. After 5 days, wild-type explants (K; n = 3) appeared consistently larger than the Vangl2 Lp/Lp organ (L; n = 3). The ureteric tree in mutants appeared distorted with wider branch tips. Balls of WT-1-expressing cells, primitive glomeruli, formed in both genotypes. Scale bars: (A and B) 50 μ m , (C and D) 125 μ m , (F) 62.5 μ m , (G–I) 63 μ m and (K and L) 125 μ m .

Article Snippet: At this stage, wild-types had significantly more ureteric bud branch tips versus Vangl2 Lp/Lp organs (Fig. J).

Techniques: In Vivo, In Vitro, Staining, Comparison, Mutagenesis, Cell Culture, Expressing

Journal: Human Molecular Genetics

Article Title: The planar cell polarity gene Vangl2 is required for mammalian kidney-branching morphogenesis and glomerular maturation

doi: 10.1093/hmg/ddq397

Figure Lengend Snippet: PCP pathway components are expressed in E18.5 kidney. Immunostaining (brown) for Scrib ( A, C and E ) and Celsr1 ( B, D and F ) in E18.5 sections (note there is no counterstain in these sections). Scrib expression was prominent in the outer (nephrogenic) cortex ( c in A) where it was found in diverse structures such as ureteric bud tips, mesenchymal condensates and vesicles (C). Scrib was also expressed in maturing podocytes located in the periphery of the glomerular tuft (E). Celsr1 expression was noted in collecting duct stalks and S-shaped bodies just below the nephrogenic zone (B and D). Expression of Celsr1 was also observed in proximal tubule and podocyte epithelia (F). Where primary antibodies were omitted, no significant staining was observed ( G and H ). Dashed line in (G) marks edge of kidney; below line is tissue. c , cortex; g , glomerulus; m , medulla; pt, proximal tubule. Scale bars: (A and B) 100 μ m , (C and D), (G) 25 μ m and (E, F and H) 12.5 μ m .

Article Snippet: At this stage, wild-types had significantly more ureteric bud branch tips versus Vangl2 Lp/Lp organs (Fig. J).

Techniques: Immunostaining, Expressing, Staining