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nrp1xenopus   

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Experiment details for nrp1

Borchers AG et al. (2002) Assay

The E3 ubiquitin ligase GREUL1 anteriorizes ectoderm during Xenopus development.

Gene Clone Species Stages Anatomy
nrp1.L laevis NF stage 19 to NF stage 21 neural plate

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  FIG. 1. GREUL1 converts epidermis into cement gland and neural tissue in whole embryos. Embryos were injected with 500 pg of GREUL1 mRNA into one blastomere of two-cell Xenopus embryos. At late neurula stage (19�21), the embryos were stained by in situ hybridization for tissue-specific markers. (A, B) Laterally viewed, XAG-1-stained embryos, injected (A) compared with uninjected (B). The inset in (B) shows an anterior view of the same embryo. (C, D) Laterally viewed, Xotx2-stained embryos, injected (C) compared with uninjected (D). (E) Dorsally viewed, injected and Nrp1-stained embryos. The injected side, marked by an arrow, can be clearly distinguished from the uninjected side. (F) Embryos injected at the one-cell stage with 500 pg of GREUL1 and stained for N-tubulin. (G) 500 pg GREUL1-injected embryos stained for c-actin, showing normal somite development. In (H), the embryos were also injected with lacZ and stained for -galactosidase activity (red) prior to in situ hybridization for slug. Dashed lines separate control and GREUL1/lacZ-injected sides. The control side is oriented toward the top. (I�L) One-cell-stage GREUL1-injected embryos stained by in situ hybridization for both XAG-1 (magenta) and GREUL1 (blue�green). (I) was injected with 1 ng of GREUL1, and (J, K) were injected with 500 pg of GREUL1. (L) An uninjected control. (M) An expanded view of a portion of the embryo in (K), showing a few ectopic XAG-1 dots that appear to be outside of the blue�green GREUL1-expressing region.

Gene Clone Species Stages Anatomy
nrp1.L laevis NF stage 28 brain , eye , anterior neural tube , neural tube

  FIG. 3. GREUL1 makes ectoderm competent to respond to neural-inducing signals. (A) Animal cap explants were cut at stage 8�9 and transplanted into the lateral flanks of stage 14 embryos. At stage 28, transplants were stained with Nrp1 and lacZ. (B) GREUL1-overexpressing grafts were cut from the lateral flanks of stage 14 embryos and grafted into uninjected controls. At stage 28, the graft is visible by its punctuate pattern of Nrp1 expression. (C) GREUL1-overexpressing neural tubes were grafted into uninjected control embryos and analyzed for Nrp1 expression. Nrp1-positive cells outside the graft were not observed (red arrow).