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Experiment details for rgn

Cartry J et al. (2006) Assay

Retinoic acid signalling is required for specification of pronephric cell fate.

Gene Clone Species Stages Anatomy
rgn.L laevis NF stage 33 and 34 pronephric kidney

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  Fig. 1. Effects of citral and BMS-453 on pronephros marker genes. Embryos were incubated during gastrulation with citral (B, E) or BMS-453 (C, F) or cultured in control medium (A, D). At the late tailbud stage 33, XSMP-30 (A–C) and XPax-2 (D–F) expression was analysed by whole mount in situ hybridization. Panels D′, E′, F′ are high magnification of the pronephric tubules area of panels D, E, F, respectively. XSMP-30 expression is severely reduced in embryos treated with the chemical inhibitors (B, C). XPax-2 expression in connective tubules and nephrostomes (arrowheads in panel D′) is lost in treated embryos (E′, F′) while expression in the duct is not affected.

Gene Clone Species Stages Anatomy
rgn.L laevis NF stage 33 and 34 pronephric duct , pronephric kidney

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  Fig. 2. Ectopic expression of XCyp26 or dnRAR inhibits pronephros formation and expression of pronephros marker genes. Four-cell stage embryos were injected with XCyp26 (D–F) or dnRAR (G–I) encoding mRNA together with LacZ mRNA into the four blastomeres (A–I) or the two blastomeres fated to give rise to the left side (J–L). In the later case, the right side of the embryo is used as an internal control. A–I: In situ hybridization (purple staining) for XPax-2 (A, D, G) XSMP-30 (B, E, H) and XWT-1 (C, F, I). X-gal staining is revealed in red (D–I). In control embryos, XPax-2 is expressed in the tubules and the duct while XSMP-30 is restricted to the tubules and XWT-1 expressed in the glomus. No signal for these genes in either of theses structures was detected. J–L: Transverse histological section of tadpole stage 38. Several pronephric tubules (red circle) are clearly visible on the control side (K, and high magnification in panel J) whereas none is observed on the injected side (K, and high magnification in panel L).

Gene Clone Species Stages Anatomy
rgn.L laevis NF stage 33 and 34 glomus , pronephric kidney

  Fig. 6. RA signalling is required in pronephric precursors. A–C: XSMP-30 expression in embryos injected in the two dorsal blastomeres at the 4-cell stage with XCyp26 mRNA and β-galactosidase encoding mRNA as a lineage tracer. X-gal staining is revealed in blue and XSMP-30 expression in purple. When XCyp26 overexpression is restricted to the dorsal part of the embryos, XSMP-30 expression is not affected (B) whereas it is severely reduced when XCyp26 expression encompasses the pronephric primordium (arrowhead in panel C). An uninjected embryo is shown in panel A. D: Experimental design for the transplantation assay. E–G: Grafted embryo cultured to the tailbud stage 28. Lateral view of the anterior region showing localization of RLDx (red, F) and GFP/dnRAR (green, G) expressing cells. H–M: Grafted embryo cultured to the tadpole stage 38. Lateral view of the anterior region showing localization of RLDx (red, I) and GFP/dnRAR (green, J). Panels K, L and M present the same transverse histological section in the pronephric tubules area showing dapi staining (K), RLDx (L) and GFP (M) localization. Tubules contain RLDx but not GFP/dnRAR cells. In panels E–J, anterior is on the right and dorsal up.

Gene Clone Species Stages Anatomy
rgn.L laevis NF stage 33 and 34 pronephric kidney

  rpn (regucalcin) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 33 & 34, lateral view, anterior right, dorsal up.

Gene Clone Species Stages Anatomy
rgn.L laevis NF stage 40 pronephric nephron , glomus , pronephric kidney

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  Fig. 4. Increased RA signalling expands the size of the pronephros. Four-cell stage embryos were co-injected with β-galactosidase encoding mRNA and XRaldh2 (A–D) or VP16-xRARα1 (E, F) mRNAs into the two blastomeres fated to give rise to the left side (right column). Embryos in panels A–D were treated 30 min with ATR. XPax-8 (A, B, E, F) and XSMP-30 expression (C, D) was analysed by in situ hybridization at the early tailbud and tadpole stages respectively (purple staining). X-gal staining is revealed in red (B, D) or light blue (F). Panels A′, B′, C′, D′ are pronephros area enlargements of panels A, B, C, D, respectively. XPax-8 and XSMP-30 expression in the pronephros primordium is enlarged on the injected side compare to the uninjected one.