Relationship of vegetal cortical dorsal factors in the Xenopus egg with the Wnt/ β-catenin signaling pathway

Abstract

In Xenopus, the dorsal factor in the vegetal cortical cytoplasm (VCC) of the egg is responsible for axis formation of the embryo. Previous studies have shown that VCC dorsal factor has properties similar to activators of the Wnt/ β-catenin-signaling pathway. In this study, we examined the relationship of the VCC dorsal factor with components of the pathway. First, we tested whether β-catenin protein, which is known to be localized on the dorsal side of early embryos, accounts for the dorsal axis activity of VCC. Reduction of β-catenin mRNA and protein in oocytes did not diminish the activity of VCC to induce a secondary axis in recipient embryos. The amount of β-catenin protein was not enriched in VCC compared to animal cortical cytoplasm, which has no dorsal axis activity. These results indicate that β-catenin is unlikely to be the VCC dorsal axis factor. Secondly, we examined the effects of four Wnt-pathway-interfering constructs (dominant-negative Xdsh, XGSK3, Axin, and dominant-negative XTcf3) on the ability of VCC to induce expression of the early Wnt target genes, Siamois and Xnr3. The activity of VCC was inhibited by Axin and dominant negative XTcf3 but not by dominant negative Xdsh or XGSK3. We also showed that VCC decreased neither the amount nor the activity of exogenous XGSK3, suggesting that the VCC dorsal factor does not act by affecting XGSK3 directly. Finally, we tested six Wnt-pathway activating constructs ( Xwnt8, Xdsh, dominant negative XGSK3, dominant negative Axin, XAPC and β-catenin) for their responses to the four Wnt-pathway-interfering constructs. We found that only XAPC exhibited the same responses as VCC; it was inhibited by Axin and dominant negative XTcf3 but not by dominant negative Xdsh or XGSK3. Although the connection between XAPC and the VCC dorsal factor is not yet clear, the fact that APC binds Axin suggests that the VCC dorsal factor could act on Axin rather than XGSK3.