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Experiment details for aplnr

Kazanskaya O et al. (2008) Assay

The Wnt signaling regulator R-spondin 3 promotes angioblast and vascular development.

Gene Clone Species Stages Anatomy
aplnr tropicalis NF stage 24 mesoderm , ventral blood island , epicardial precursor cell

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  Fig. 1. Rspo3 is expressed in endothelial cells and their precursors. (A-K) Rspo3 expression by in situ hybridization during mouse and Xenopus embryogenesis. (A-D) Mouse embryo. (E-H) X. laevis. (I-K) X. tropicalis. (A) E8.0 embryo; anterior towards the left. Rspo3 expression is in the primitive streak and in the allantois. (B) E8.5 embryo; anterior towards the top. Rspo3 expression is prominent in the posterior mesoderm and in the allantois. (C) Rspo3 expression in vasculature of the chorionic plate (E8.5). (D) Expression in the umbilical vein (arrowhead) (E12.5). (E) Stage 17; dorsal view, anterior leftwards, showing expression in the neural plate. (F) Stage 28; lateral view with anterior leftwards, showing expression in organs labeled. (G,H) Transverse section at tailbud stage showing expression of Rspo3 (G) and Vegf (H) in the hypochord (arrowheads). (I-K) Expression of indicated genes in stage 24; ventral view, anterior leftwards, showing expression of Msr and Rspo3 in the periphery of the ventral blood islands. Abbreviations: all, allantois; ba, branchial arches; cp, chorionic plate; fb, forebrain; dnt, dorsal neural tube; dlp, dorsal lateral plate; lb, limb bud; n, notochord; pm, posterior mesoderm; pn, pronephric sinus; ps, primitive streak; tb, tailbud mesoderm; vbi, ventral blood island.

Gene Clone Species Stages Anatomy
aplnr tropicalis NF stage 28 ventral blood island , blood , splanchnic layer of lateral plate mesoderm , lateral plate mesoderm , blood vessel , [+]

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  Fig. 2. Rspo3 regulates the balance between blood and endothelial differentiation. (A-D) Rspo3 is required for blood vessel formation. Xenopus tropicalis embryos were injected equatorially into each blastomere at the four-cell stage with 2.5 ng control morpholino (Co Mo) or Rspo3 Mo as indicated. (A,B) Tadpole morphology; note edema in embryo injected with Rspo3 Mo (B; 69.0%, n=203) but not in control (A) (6.4%, n=173). (C,D) o-Dianisidine erythrocyte staining at tadpole stage. Poor blood vessel formation occurs in the head and erythroid cells accumulate in the ventral region with Rspo3 Mo (D). (E-L) Rspo3 is required for angioblast formation. Xenopus tropicalis four-cell stage embryos were injected equatorially with 2.5 ng Co Mo or Rspo3 Mo, as indicated, into each blastomere and analyzed at tailbud stage by whole-mount in situ hybridization for the indicated genes. (H,L) Inset shows high magnification of the lateral region with Msr-expressing angioblasts. Control and Rspo3 Mo showed Msr staining at 93% (n=75) and 20% (n=87) frequency, respectively. (M-O) Rescue experiment of Rspo3 Mo-induced phenotype. Xenopus tropicalis four-cell stage embryos were injected with 2.5 ng of Co Mo or Rspo3 Mo with or without 50 pg Xenopus Rspo2 mRNA, as indicated, into each ventral blastomere. At gastrula stage (st. 10.5), ventral marginal zones (VMZ) were explanted, cultured until stage 28 and processed for whole-mount in situ hybridization for α-globin. The upregulation ofα -globin expression (23/24) in Rsop3 Mo-injected VMZ and the downregulation of α-globin expression (19/20) in Rspo3 Mo and Rspo2 mRNA-injected VMZ was observed in two independent experiments. (P-W) BSA-, Rspo2- or VEGF-soaked beads (top label) were transplanted in the ventral blood island precursors in neurulae and processed for whole-mount in situ hybridization for the indicated markers (side label) at tadpole stage. Arrowheads indicate ectopic Msr induction. White circles indicate implanted beads.