Immunocytochemical studies of the interactions of cadherins and catenins in the early Xenopus embryo.

Linkage of cadherins to the cytoskeleton is crucial for their adhesive function. Since alpha- and beta-catenin play a key role in this linkage, these proteins are possible targets for processes that control cell-cell adhesion. To achieve a better understanding of the regulation of cell-cell adhesion in embryonic morphogenesis, we used immunohistology to investigate how in Xenopus blastomeres catenins respond to disturbances in the expression of maternal cadherins. Overexpression of myc-tagged maternal cadherin leads to a proportionate increase of the level of beta-catenin. The two proteins colocalize in the endoplasmic reticulum, in cytoplasmic vesicles, and along the cell membrane, indicating that the beta-catenin binds to overexpressed cadherin early in its passage to the plasma membrane. Expression of cadherin is essential for the stable presence of beta-catenin, as depletion from maternal cadherin mRNA leads to a complete loss of beta-catenin from the blastomeres. alpha-Catenin behaves differently. Overexpression of cadherin leaves the amount and localization of alpha-catenin largely unaffected, and additional cadherin inserts itself into the membrane without a proportionate rise in the level of membrane-bound alpha-catenin. However, cadherin mRNA depletion leads to a redistribution of alpha-catenin from the membrane to the cytoplasm. Thus, cadherin is required to localize alpha-catenin to the membrane, but the amount of alpha-catenin along the membrane seems to be restricted to a certain level which cannot be exceeded. The relevance of these observations for the regulation of cadherin-mediated cell adhesion in the Xenopus embryo is discussed. Additionally, we demonstrate that plakoglobin, like beta-catenin an armadillo repeat protein, shows neither accumulation after overexpression nor colocalization with the overexpressed cadherin.

A paired oocyte adhesion assay reveals the homophilic binding properties of the Xenopus maternal cadherins, XB/U- and EP-cadherin.

The homophilic nature of cadherin-mediated cell-cell adhesion provides an organism with the opportunity of altering the adhesive capabilities of its cells by selectively modulating the expression of different cadherin types. Differential cadherin expression is of major importance in regulating the cell rearrangements involved in the processes which shape tissues and organs during embryogenesis. The pregastrula embryo of Xenopus laevis expresses two maternally supplied cadherins: XB/U-cadherin and EP-cadherin. Since these two proteins are almost 92% identical at the amino acid level, it was unclear whether heterophilic interactions between them were possible. Different functional roles can only be ascribed to the two cadherins if the possibility of heterophilic binding between them can be excluded. We describe a simple and straightforward assay which can be used to assess interactions between adhesion molecules. A combination of antisense oligonucleotide and enzyme treatments eliminates endogenous cadherins in Xenopus oocytes and subsequent injection of a specific mRNA yields oocytes carrying only one or the other cadherin. After removal of the vitelline membranes, two oocytes expressing the appropriate cadherins will adhere to one another when they are placed in close contact. By scoring for adhesion in homotypic and heterotypic pairings, we demonstrate that XB/U-cadherin and EP-cadherin do not interact with one another.