The role of cadherins during primordial germ cell migration and early gonad formation in the mouse.

Primordial germ cells (PGCs) are the founder cells of the gametes. In mammals, PGCs migrate from the hindgut to the genital ridges, where they coalesce with each other and with somatic cells to form the primary sex cords. We show here that, in both sexes, PGCs express P- and E-cadherins during and after migration, and N-cadherin at post-migratory stages. E-Cadherin is not expressed by PGCs whilst in the hindgut, but is upregulated as they leave. Blocking antibodies against E-, but not P-cadherin cause defective PGC-PGC coalescence, and in some cases, ectopic PGCs.

A beta-catenin/XTcf-3 complex binds to the siamois promoter to regulate dorsal axis specification in Xenopus.

The Wnt pathway regulates the early dorsal-ventral axis in Xenopus through a complex of beta-catenin and HMG box transcription factors of the Lef/Tcf family. We show that the promoter of the dorsalizing homeo box gene siamois is a direct target for the beta-catenin/XTcf-3 complex, establishing a link between the Wnt pathway and the activation of genes involved in specifying the dorsal axis. By injecting siamois reporter constructs into the animal pole of Xenopus embryos, we show that a 0.8-kb fragment of the siamois promoter is strongly activated by beta-catenin. The proximal 0.5 kb, which is also activated by beta-catenin, contains three Lef/Tcf-binding sites. Mutations in these sites eliminate the beta-catenin-mediated activation of siamois and show that siamois is regulated by the beta-catenin/XTcf-3 complex, in combination with additional transcriptional activators. When expressed at the equator of the embryo, the siamois promoter is activated to much higher levels on the dorsal side than the ventral side. Ectopic ventral expression of beta-catenin raises the ventral expression of the siamois promoter to the dorsal levels. Conversely, ectopic dorsal expression of dominant-negative XTcf-3 abolishes the dorsal activation of the siamois promoter. Furthermore, elimination of the Lef/Tcf sites elevates the ventral expression of siamois, revealing a repressive role for XTcf-3 in the absence of beta-catenin. Finally, we find that the endogenous siamois activator, although present throughout the dorsal side of the embryo, is most potent in the dorsal vegetal region. We propose that the dorsal activation of siamois by the beta-catenin/XTcf-3 complex combined with the ventral repression of siamois by XTcf-3 results in the restriction of endogenous siamois expression to the dorsal side of Xenopus embryos.

Interactions between primordial germ cells play a role in their migration in mouse embryos.

Primordial germ cells (PGCs) are the founder cell population of the gametes which form during the sexually mature stage of the life cycle. In the mouse, they arise early in embryogenesis, first becoming visible in the extraembryonic mesoderm, posterior to the primitive streak, at 7.5 days post coitum (d.p.c.). They subsequently become incorporated into the epithelium of the hind gut, from which they emigrate (9.5 d.p.c.) and move first into the dorsal mesentery (10.5 d.p.c.), and then into the genital ridges that lie on the dorsal body wall (11.5 d.p.c.). We have used confocal microscopy to study PGCs stained with an antibody that reacts with a carbohydrate antigen (Stage-Specific Embryonic Antigen-1, SSEA-1) carried on the PGC surface. This allows the study of the whole PGC surface, at different stages of their migration. The appearance of PGCs in tissue sections has given rise to the conventional view that they migrate as individuals, each arriving in turn at the genital ridge. In this paper, we show that PGCs leave the hind gut independently, but then extend long (up to 40 microns) processes, with which they link up to each other to form extensive networks. During the 10.5-11.5 d.p.c. period, these networks of PGCs aggregate into groups of tightly apposed cells in the genital ridges. As this occurs, their processes are lost, and their appearance suggests they are now non-motile. Furthermore, we find that PGCs taken from the dorsal mesentery at 10.5 d.p.c. perform the same sequence of movements in culture.(ABSTRACT TRUNCATED AT 250 WORDS)

Primordial germ cell migration.

Primordial germ cells are migratory cells. They arise very early in embryogenesis and have a similar pattern of migration in Drosophila, Xenopus, chick and mouse. In each case the primordial germ cells associate with the developing gut from which they migrate to the gonads during organogenesis. Germ cells proliferate mitotically from the time they begin to migrate to the time they colonize the genital ridges. From the study of mouse primordial germ cells, we now know of a number of agents which affect primordial germ cell proliferation, migration and adhesion in vitro. More recently, we have studied the interactions between primordial germ cells and the cells and extracellular matrix molecules on their migratory route. By labelling germ cells in whole-mount preparations with an antibody to the germ cell marker SSEA-1, we have studied the spatial distribution of germ cells in situ using confocal microscopy. This study has revealed that germ cells link up with each other forming extensive networks during migration.

Mitogen-induced expression of the primary response gene cMG1 in a rat intestinal epithelial cell-line (RIE-1).

cMG1 is a primary response gene first identified in a rat intestinal epithelial (RIE-1) cell-line [(1990) Oncogene 5, 1081-1083]. A number of mitogens, including epidermal growth factor (EGF), angiotensin II (AII), serum and insulin rapidly induced 2- to 6-fold increases of cMG1 mRNA in RIE-1 cells, while transforming growth factor-beta caused a small reduction. Cyclic AMP-elevating agents blocked the increase of cMG1 mRNA induced by EGF. The AII-stimulated increase of cMG1 mRNA was blocked by the depletion of protein kinase C, whereas the EGF-stimulated increase was not affected, indicating that protein kinase C-dependent and -independent signalling pathways stimulate cMG1 expression.

The nucleotide sequence of a cDNA encoding an EGF-inducible gene indicates the existence of a new family of mitogen-induced genes.

A gene, cMG1, whose expression is transiently activated in growth factor stimulated epithelial cells has been identified by differential screening. Overlapping cDNA recombinants derived from cMG1 mRNA have been sequenced and shown to encode a protein of 338 amino acids. Database searches have revealed no similarities between cMG1 and other genes, except that over a 67-amino acid region the cMG1 protein shows 72% identity to the product of the TIS11 gene, a TPA-induced sequence in Swiss 3T3 cells. The results suggest that these two genes code for a novel type of early response gene.

Periodic DNA synthesis in cell-free extracts of Xenopus eggs.

Cell-free extracts prepared from unfertilized eggs of Xenopus laevis support DNA synthesis on sperm pronuclei. Continuous labelling studies using [3H]dCTP and pulse labelling studies using [32P]dCTP demonstrate that synthesis occurs in short bursts of 40 min, which are punctuated by periods of 20-40 min during which no synthesis occurs. Density substitution experiments using bromodeoxyuridine demonstrate that this synthesis involves the initiation of replication and reveals that re-initiation events can occur following multiple bursts of replication. The periodic properties of these extracts are sensitive to protein synthesis inhibitors.