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Experiment details for pax8

Cartry J et al. (2006) Assay

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

Gene Clone Species Stages Anatomy
pax8.L laevis NF stage 12 mesoderm , axial mesoderm , pronephric mesenchyme , lateral

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  Fig. 8. RAR-α is expressed in pronephric mesoderm. A: RT-PCR analysis of RAR-β expression during Xenopus embryogenesis. Embryonic stages are indicated according to Nieuwkoop and Faber, 1967. ODC is used as an RNA loading control. B–L: In situ hybridization on gastrula stage 11 (B–G) or stage 12 (H–L) for XRaldh2 (B, E, J), RAR-γ (C, F, K), RAR-α (D, G, L), XPax-8 (H) and Xlim-1 (I). In situ hybridization was performed on hemisectioned embryos except in panels B–D. In panels H–L, embryos were cut transversally at the level of the pronephric territory, dorsal is up. RAR-α and XRaldh2 are expressed in the mesodermal layer during gastrulation and their expression domains overlap with Xlim-1 and XPax-8 expression in the pronephric territory at the late gastrula stage.

Gene Clone Species Stages Anatomy
pax8.L laevis NF stage 12 mesoderm , pronephric mesenchyme , lateral
pax8.L laevis NF stage 17 mesoderm , pronephric mesenchyme , lateral

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  Fig. 10. Xlim-1 and XPax-8 expression at the late gastrula stage depends on RA signalling and RA induces Xlim-1 expression in absence of protein synthesis. A–H: In situ hybridization for Xlim-1 (A–D) and XPax-8 (E–H) in control embryos (A, B, E, F) or embryos incubated with BMS-453 (C, D, G, H) for 6 h since the gastrula stage 10.5 (C, G) or the neurula stage 17 (D, H). Xlim-1 and XPax-8 staining in the pronephric lineage is reduced when the inhibitor treatment is applied at gastrula stage but not at neurula stage. I–P: Embryos incubated with RA (1 μM) at the late gastrula stage 12.5 and analysed for Xlim-1 (I–N) or XPax-8 (O, P) expression by in situ hybridization 20, 40, 60 or 120 min after the onset of the treatment. Q–T: Xlim-1 expression in control embryos (Q), in embryos treated at the late gastrula stage 12.5 with CHX (R), RA (S) or RA and CHX (T). CHX was applied 30 min before the onset of the 2 h RA treatment.

Gene Clone Species Stages Anatomy
pax8.L laevis NF stage 13 mesoderm , pronephric mesenchyme , lateral

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  Fig. 9. Ectopic XCyp26 mRNA affects early expression of XPax-8 and Xlim-1. Four-cell stage embryos were injected with XCyp26 and β-galactosidase encoding mRNAs into the two blastomeres fated to give rise to the left side. At stage 13, Xlim-1 and XPax-8 expression was analysed by combined in situ hybridization (purple staining) and β-galactosidase activity (red staining). Xlim-1 (A, B) and XPax-8 (C, D) expression in the area fated to give pronephros is severely reduced on the injected side. Note that expression of XPax-8 in the otic placode is also reduced. Dorsal is up, anterior is to the right for the control sides and to the left for the injected sides.

Gene Clone Species Stages Anatomy
pax8.L laevis NF stage 22 otic placode , pronephric mesenchyme

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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.