Involvement of a neutral glycolipid in differential cell adhesion in the Xenopus blastula.

Many different molecular species mediate cell adhesion during embryonic development. These can have either protein or carbohydrate functional groups, which can act in either a homophilic or a heterophilic manner, and often in concert. We report here that a monoclonal antibody, M4B, raised against Xenopus blastomere membranes, inhibits the calcium-dependent adhesion of dissociated blastomeres.

Function of maternal cytokeratin in Xenopus development.

Intermediate filaments are ubiquitous in eukaryotic cells, but their functions are poorly understood. The Xenopus oocyte contains both messenger RNA and protein products of cytokeratin and vimentin genes in non-overlapping arrays. The cytokeratin filaments contain dimers of the type I (acidic) subunit XLK3a(19), and the type II (basic) subunit XCK1(8), polymerized to form a cortical network.

Antipredator Defenses in Tropical Pacific Soft Corals (Coelenterata: Alcyonacea). I. Sclerites as Defenses Against Generalist Carnivorous Fishes.

Calcified sclerites are common in many invertebrate species and are frequently used as taxonomic indicators; however, little is known about the function of sclerites. To determine whether sclerites could function as antipredator defenses, we conducted field assays in which sclerites from the Indo-Pacific soft corals Sinularia maxima, S. polydactyla, and S.

Distinct distribution of vimentin and cytokeratin in Xenopus oocytes and early embryos.

We report the identity of a major component of Triton-insoluble extracts from Xenopus oocytes and early embryos. In a previous paper we showed that an antibody, Z9, cross-reacts with two polypeptides from such extracts (Mr 56,000 and 57,000) as well as Xenopus vimentin. Direct microsequencing of the Mr 57,000 protein shows near identity of three tryptic fragments with regions of the predicted amino acid sequence of XCK1(8), a basic cytokeratin whose mRNA is known to be expressed in Xenopus oocytes.

The role of intermediate filaments in early Xenopus development studied by antisense depletion of maternal mRNA.

The effects of depleting a maternal cytokeratin mRNA on the developing embryo are described. Cytokeratins are members of the intermediate filament family of cytoskeletal proteins, and are expressed in a cortical network of the superficial cytoplasm of the oocyte. After fertilisation, a new cortical network is built up, which comes to occupy only the most superficial cells of the blastula.

TGF beta 1 inhibits proliferation and has a chemotropic effect on mouse primordial germ cells in culture.

Primordial germ cells are the stem cells that provide the functional gametes of adult animals. In many animal groups they are set aside at the earliest stages of development, and migrate from their sites of first appearance to the sites where the gonad will form, the genital ridges. During this migration they proliferate.

Response to fibronectin of mouse primordial germ cells before, during and after migration.

The adhesive extracellular matrix glycoprotein fibronectin is thought to play a central role in cell migration during embryogenesis. In order to define this role, we have examined the response to fibronectin in cell culture of mouse primordial germ cells (PGCs) before, during and after their migration from the hindgut into their target tissue, the genital ridges. Using an explant culture system, we show that PGCs will emigrate from tissue fragments containing hindgut, and that fibronectin stimulates this migration.

Effects of the steel gene product on mouse primordial germ cells in culture.

Mutations at the steel (sl) and dominant white spotting (W) loci in the mouse affect primordial germ cells (PGC), melanoblasts and haemopoietic stem cells. The W gene encodes a cell-surface receptor of the tyrosine kinase family, the proto-oncogene c-kit. In situ analysis has shown c-kit messenger RNA expression in PGC in the early genital ridges.

Fertilization of cultured Xenopus oocytes and use in studies of maternally inherited molecules.

The methods described here of fertilizing stage VI oocytes are lengthy and quite difficult techniques. They would become more attractive if the success rate (i.e.

Identification of vimentin and novel vimentin-related proteins in Xenopus oocytes and early embryos.

We have made antibodies against fusion proteins of Xenopus vimentin. We show for the first time the distribution of vimentin in larval stages, where it is found in cells of mesenchymal origin, and in radial glial cells. In sections of Xenopus oocytes and early embryos, immunocytochemistry reveals the presence of an extensive cytoplasmic network, distributed in an animal-vegetal gradient.

Genital ridges exert long-range effects on mouse primordial germ cell numbers and direction of migration in culture.

The functional gametes of all vertebrates first arise in the early embryo as a migratory population of cells, the primordial germ cells (PGCs). These migrate to, and colonise, the genital ridges (GR) during the early organogenesis period, giving rise to the complete differentiating gonad. PGCs first become visible by alkaline phosphatase staining in the root of the developing allantois at 8.

Regional identity is established before gastrulation in the Xenopus embryo.

Regional differences in cell recognition properties along the animal-vegetal axis of Xenopus laevis embryos were investigated by using an in vitro cell sorting assay. Dissociated cells were obtained from defined regions of blastula- and early gastrula-stage embryos. Binary combinations of cells from different regions, or from the same region at different ages, were aggregated in stationary culture.

The establishment of regional identity in the Xenopus blastula.

The molecules involved in the commitment of Xenopus cells to particular germ layers are unknown. The question has been investigated for the cells of the blastula in in vivo cell transplantation assays and in vitro aggregation assays. Using the former technique, we have shown that vegetal cells become committed before gastrulation, even when placed in inappropriate sites.

Vimentin expression in oocytes, eggs and early embryos of Xenopus laevis.

Immunocytochemical studies using a monoclonal anti-porcine vimentin antibody reveal a well-organized pattern of staining in Xenopus laevis oocytes, eggs and early embryos. The positions of Xenopus vimentin and desmin in two-dimensional (2D) polyacrylamide gels were first established by immunoblotting of muscle Triton extracts with anti-intermediate filament antibodies (anti-IFA), which cross-react with all intermediate filament proteins (IFPs). The anti-porcine vimentin reacts with vimentin and desmin in muscle 2D immunoblots, but only reacts with one polypeptide in oocyte blots in the position predicted for vimentin (Mr 55 x 10(3), pI 5.

Changes in states of commitment of single animal pole blastomeres of Xenopus laevis.

The experiments described in this paper were designed to compare the normal fates of animal pole blastomeres of Xenopus laevis with their state of commitment. Single animal pole blastomeres were labeled with a lineage marker and transplanted into the blastocoels of host embryos of different stages. The distribution of labeled daughter cells in the tadpole reflects the state of commitment of the parent cell at the time of transplantation.

Vegetal pole cells and commitment to form endoderm in Xenopus laevis.

In order to compare their states of commitment with their normal developmental fate, single vegetal pole cells from early Xenopus embryos were labeled and transplanted into the blastocoels of host embryos. In a previous study we showed, using this single cell transplantation assay, that vegetal pole cells become committed to endoderm by the early gastrula stage. In this paper we examine some properties of the commitment process.

Studies on the migration of mouse germ cells.

Primordial germ cells in the mouse embryo migrate from their site of origin to the gonad where they differentiate, giving rise eventually to the gametes of the mature adult animal. The migratory phase is transient and therefore permits analysis of factors regulating the motile activity of cells in tissues. Germ cells can be isolated during migration and cultured on feeder cells of an established cell line (STO).

Invasive behaviour of mouse primordial germ cells in vitro.

We have isolated migrating primordial germ cells (PGCs) from 10.5-day mouse embryos and studied their behaviour when cultured on a mouse embryo fibroblast (STO) cell line. Living and fixed PGCs were identified by fluorescent labelling with a monoclonal antibody specific for PGCs in the culture system used.

The appearance and distribution of intermediate filament proteins during differentiation of the central nervous system, skin and notochord of Xenopus laevis.

Antibodies against various intermediate filament proteins have been used to follow cell differentiation in the early Xenopus embryo. Three new monoclonal antibodies against Xenopus cytokeratins raised against Triton-insoluble material from tadpoles (RD35/2a, RD35/3a and D3/3a), two antibodies against mammalian cytokeratins (LE65 and LP3K), monoclonal anti-(rat 200 K neurofilament protein), rabbit anti-(rat glial filament acidic protein), and rabbit antibodies to hamster and calf vimentin were used. We show that cytokeratins are present in the early central nervous system (CNS) and persist in the ependymal cells of the adult CNS.

Cinemicrographic study of the cell movement in the primitive-streak-stage mouse embryo.

Migration of the mesoderm cells in the primitive-streak-stage mouse embryo was directly studied by cinemicrography using whole embryo culture and Nomarski differential interference contrast optics. Relative transparency and small size of the early mouse embryos enabled direct observation of the individual cells and their cell processes. Seven-day-old mouse embryos were isolated and cultured in a small chamber in a medium consisting of 50% rat serum and 50% Dulbecco's modified minimum essential medium.

Migratory and postmigratory mouse primordial germ cells behave differently in culture.

In all vertebrate groups, the progenitors of the germ line, the primordial germ cells (PGCs) arise extragonadally and move to the developing gonad early in embryonic development. We have examined the behavior of isolated pregonadal and gonadal PGCs in vitro on feeder layers of an embryo-derived cell line. Histochemically and serologically identified pregonadal germ cells are found to be actively motile in vitro and, furthermore, show behavior characteristic of invasive cells.