Zhang B et al. (2011), Expression of Wnt signaling components during X...

XB-ART-44122

PLoS One 2011 Jan 01;610:e26533. doi: 10.1371/journal.pone.0026533.

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Expression of Wnt signaling components during Xenopus pronephros development.

Zhang B , Tran U , Wessely O .

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The formation of the vertebrate kidney is tightly regulated and relies on multiple evolutionarily conserved inductive events. These are present in the complex metanephric kidney of higher vertebrates, but also in the more primitive pronephric kidney functional in the larval stages of amphibians and fish. Wnts have long been viewed as central in this process. Canonical β-Catenin-dependent Wnt signaling establishes kidney progenitors and non-canonical β-Catenin-independent Wnt signaling participate in the morphogenetic processes that form the highly sophisticated nephron structure. While some individual Wnt signaling components have been studied extensively in the kidney, the overall pathway has not yet been analyzed in depth. Here we report a detailed expression analysis of all Wnt ligands, receptors and several downstream Wnt effectors during pronephros development in Xenopus laevis using in situ hybridization. Out of 19 Wnt ligands, only three, Wnt4, Wnt9a and Wnt11, are specifically expressed in the pronephros. Others such as Wnt8a are present, but in a broader domain comprising adjacent tissues in addition to the kidney. The same paradigm is observed for the Wnt receptors and its downstream signaling components. Fzd1, Fzd4, Fzd6, Fzd7, Fzd8 as well as Celsr1 and Prickle1 show distinct expression domains in the pronephric kidney, whereas the non-traditional Wnt receptors, Ror2 and Ryk, as well as the majority of the effector molecules are rather ubiquitous. In addition to this spatial regulation, the timing of expression is also tightly regulated. In particular, non-canonical Wnt signaling seems to be restricted to later stages of pronephros development. Together these data suggest a complex cross talk between canonical and non-canonical Wnt signaling is required to establish a functional pronephric kidney.

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Species referenced: Xenopus laevis
Genes referenced: celsr1 celsr2 celsr3 ctnnb1 dvl1 dvl2 dvl3 fzd1 fzd10 fzd2 fzd3 fzd4 fzd5 fzd6 fzd7 fzd8 fzd9 prickle1 prickle2 prickle3 ror2 ryk vangl1 vangl2 wnt1 wnt10a wnt10b wnt11 wnt11b wnt16 wnt2b wnt3a wnt4 wnt5a wnt5b wnt6 wnt7a wnt7b wnt7c wnt8a wnt8b wnt9a wnt9b wnt9b.2

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	Figure 1. Expression of Wnt Ligands at Stage 25. Whole mount in situ hybridization of Xenopus embryos at stage 25 for Wnt1 (A), Wnt2b (B), Wnt3a (C), Wnt4 (D), Wnt5a (E), Wnt5b (F), Wnt6 (G), Wnt7a (H), Wnt7b (I), Wnt7c (J), Wnt8a (K), Wnt8b (L), Wnt9a (M), Wnt9b (N), Wnt10a (O), Wnt10b (P), Wnt11 (Q), Wnt11b (R) and Wnt16 (S). Inset in D shows close-up of the pronephric anlage. All images are of the same magnification and the scale bar corresponds to 0.5 mm.
	Figure 2. Expression of Wnt Ligands at Stage 35. Whole mount in situ hybridization of Xenopus embryos at stage 35 for Wnt1 (A), Wnt2b (B), Wnt3a (C), Wnt4 (D), Wnt5a (E), Wnt5b (F), Wnt6 (G), Wnt7a (H), Wnt7b (I), Wnt7c (J), Wnt8a (K), Wnt8b (L), Wnt9a (M), Wnt9b (N), Wnt10a (O), Wnt10b (P), Wnt11 (Q), Wnt11b (R) and Wnt16 (S). Insets in D and M show close-ups of the pronephric tubular region. All images are of the same magnification and the scale bar corresponds to 0.5 mm.
	Figure 3. Expression of Wnt Receptors at Stage 25. Whole mount in situ hybridization of Xenopus embryos at stage 25 for Fzd1 (A), Fzd2 (B), Fzd3 (C), Fzd4 (D), Fzd5 (E), Fzd6 (F), Fzd7 (G), Fzd8 (H), Fzd9 (I), Fzd10 (J), Ror2 (K) and Ryk (L). All images are of the same magnification and the scale bar corresponds to 0.5 mm.
	Figure 4. Expression of Wnt Receptors at Stage 35. Whole mount in situ hybridization of Xenopus embryos at stage 35 for Fzd1 (A), Fzd2 (B), Fzd3 (C), Fzd4 (D), Fzd5 (E), Fzd6 (F), Fzd7 (G), Fzd8 (H), Fzd9 (I), Fzd10 (J), Ror2 (K) and Ryk (L). Inset in A, F and H show close-ups of the pronephric tubular region. All images are of the same magnification and the scale bar corresponds to 0.5 mm.
	Figure 5. Expression of Wnt Signaling Intermediates at Stage 25. Whole mount in situ hybridization of Xenopus embryos at stage 25 for Dvl1 (A), Dvl2 (B), Dvl3 (C), Prickle1 (D), Prickle2 (E), Prickle3 (F), Celsr1 (G), Celsr2 (H), Celsr3 (I), Vangl1 (J) and Vangl2 (K). All images are of the same magnification and the scale bar corresponds to 0.5 mm.
	Figure 6. Expression of Wnt Signaling Intermediates at Stage 35. Whole mount in situ hybridization of Xenopus embryos at stage 35 for Dvl1 (A), Dvl2 (B), Dvl3 (C), Prickle1 (D), Prickle2 (E), Prickle3 (F), Celsr1 (G), Celsr2 (H), Celsr3 (I), Vangl1 (J) and Vangl2 (K). Inset in G shows close-up of the pronephric tubular region. All images are of the same magnification and the scale bar corresponds to 0.5 mm.
	Figure 7. Pronephros Expression of Wnt Signaling Components. Paraplast sections of Xenopus embryos processed for whole mount in situ hybridization with Wnt4 (A,A, Wnt5b (B,B, Wnt8a (C,C, Wnt9a (D,D, Wnt11 (E,E, Ryk (F,F, Fzd1 (G,G, Fzd2 (H,H, Fzd4 (I,I, Fzd6 (J,J, Fzd7 (K,K, Fzd8 (L,L, Dvl1 (M,M, Dvl2 (N,N, Prickle1 (O,O, Celsr1 (P,P, Vangl1 (Q,Q and Vangl2 (R,R at stage 25 and 35. Sections are through the tubular area of the pronephros. Closed arrowheads indicate distal tubular segments; open arrowheads point towards individual dvl1-positive cells. All panels depicting embryos at stage 25 and 35, respectively are of the same magnification; a representative scale bar corresponding to 0.1 mm is shown in panels R,R
	Figure 8. Summary of Different Wnt Signaling Expression Patterns. Schematics of stage 25 and 35 cross sections summarizing the in situ hybridization data; genes following the same expression pattern are grouped together and their respective expression domains are indicated in red (or light red in the case of weak expression). dt; distal tubules; g, glomus; pa, pronephric anlage; pt, proximal tubules; ns, nephrostomes.
	Prickle2 (prickle homolog 2) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 25, lateral view, anterior left, dorsal up.
	fzd4 gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 25, lateral view, anterior left, dorsal up.
	fzd4 gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 35 & 36, lateral view, anterior left, dorsal up.

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Angers, Proximal events in Wnt signal transduction. 2009, Pubmed