Roël G et al. (2003), Tcf-1 expression during Xenopus development.

XB-ART-5393

Gene Expr Patterns 2003 May 01;32:123-6. doi: 10.1016/s1567-133x(03)00039-5.

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Tcf-1 expression during Xenopus development.

Roël G , van den Broek O , Spieker N , Peterson-Maduro J , Destrée O .

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We report the cloning and expression of Xenopus Tcf-1. The amino acid sequence of Tcf-1 of Xenopus laevis and Xenopus tropicalis is closely related to that of chicken, mouse and man. Thus, the family of Tcf/Lef proteins in the amphibian Xenopus comprises four members as in higher vertebrates. RT-PCR analysis revealed that Tcf-1 RNA encoding a beta-catenin binding isoform is maternally present as well as throughout early development. Different transcripts are expressed by alternative splicing. In cleavage and blastula stage embryos, Tcf-1 RNA is present at high levels in the animal hemisphere. During gastrulation Tcf-1 is differentially expressed with high levels in the animal cap and most of the marginal zone except for a narrow domain around the blastopore. At neurula stages expression is predominant in the neural plate. At tailbud stages expression is localized in specific areas of the brain, in the eyes, the otic vesicle, branchial arches and head mesenchyme, somites, tailbud, pronephros and pronephric duct.

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Species referenced: Xenopus tropicalis Xenopus laevis
Genes referenced: foxg1 lef1 odc1 pax2 tbx2 tcf7

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	Fig. 3. Localization of Tcf-1 transcripts containing the beta-catenin binding domain during development of X. tropicalis. Whole-mount in situ hybridizations are shown for stage 7 (A and B), stage 10.5 (C and D), stage 18 (E and F), stage 21 (I), stage 24 (J) and stage 32 (K) X. tropicalis embryos. The probe used corresponds to the N-terminal beta-catenin binding domain of XtTcf-1. (A and B) At blastula stage high levels of Tcf-1 RNA are detected in the animal hemisphere. (B) Section of embryo shown in A, white arrow indicates staining in the roof of the blastocoel. (C) Vegetal and slightly lateral view of a stage 10.5 embryo. XTcf-1 is expressed in the animal cap and most of the marginal zone except for a narrow domain around the (future) blastopore (black arrowhead). (D) Medial view of a sagittally sectioned stage 10.5 embryo; blastopore (black arrowhead), roof of blastocoel (white arrowhead). Expression is high in the sensorial layer of the animal cap and the subepithetial layer of the marginal zone (white arrows) and low in the epithetial layer (black arrows) (Hausen and Riebesell, 1991). Upper and lower parts of the dorsal lip (Ibrahim and Winklbauer, 2001) have low levels of Tcf-1 RNA (open arrowhead), while high levels are present in the subepithelial layer. Anterior involuting mesendoderm (asterisk) is negative. Ventrally, expression of Tcf-1 is high in the subepithelial layer ending in a deep zone at the edge of the blastocoel floor. (E) At stage 18 Tcf-1 transcripts are detected in the neural plate with a discrete anterior border (white arrowhead). A low signal is present at the position of the first visceral arch (black arrowhead; cf. Fig. 3G; Heller and Brändli, 1997). (F) Posterior view of embryo shown in E; expression in the posterior end of the neural plate c.q. neural folds extends ventrally around the blastopore (open arrowhead). (G) Stage 18 embryo stained for XtTcf-1 and XPax-2. The anterior expression of Pax-2 (Heller and Brändli, 1997) overlaps with the anterior border of expression of XTcf-1 (cf. Fig. 3E). (H) Stage 18 embryo stained for XBF-1 (Bourguignon et al., 1998). The expression domain of XBF-1 extends more anteriorly (white arrowhead) than XTcf-1 expression (cf. Figs. 3E,H). (I) Stage 21 embryo, showing staining in the eye (open arrowhead), specific areas of the brain, otic vesicle (red arrowhead), pronephric anlage (white arrowhead) and somites. At later stages, 24 (J) and 32 (K), Tcf-1 continues to be expressed in the eye (open arrowhead), specific areas of the brain, otic vesicle (red arrowhead), branchial arches and head mesenchyme, pronephros (white arrowhead), pronephric duct (black arrowhead), somites and tailbud. Inset in Fig. 3J shows a close-up view of Tcf-1 expression in the pronephric duct (at the level of the black arrowhead). c.g., cement gland.
	Fig. 2. Temporal expression and alternative splicing of Tcf-1 during development of Xenopus. RT-PCR analysis of X. laevis total RNA at different stages of development. Stages are indicated on top; stage 4, 10.5, 20 and 28. Tcf-1 transcripts containing the beta-catenin binding domain (N-term.) are present maternally and continue to be expressed during development. Exon VI was only detected in X. laevis adult brain cDNA (brain, arrow). RT-PCR with ODC primers was used as loading control. −RT, RNA sample without reverse transcriptase was used as a negative control for PCR.
	tcf7 (transcription factor 7 (T-cell specific, HMG-box)) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 10.5, posterior-lateral view, dorsal up.
	tcf7 (transcription factor 7 (T-cell specific, HMG-box)) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 10.5, mid-sagittal section, medial view, dorsal up.
	tcf7 (transcription factor 7 (T-cell specific, HMG-box)) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 18, anterior view, dorsal up.
	tcf7 (transcription factor 7 (T-cell specific, HMG-box)) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 21, lateral view, dorsal up, anterior left.
	tcf7 (transcription factor 7 (T-cell specific, HMG-box)) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 24, lateral view, dorsal up, anterior left.
	tcf7 (transcription factor 7 (T-cell specific, HMG-box)) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 32, lateral view, dorsal up, anterior left.