Rudolf Winklbauer

Frizzled-7 signalling controls tissue separation during Xenopus gastrulation

Cell signalling through Frizzled receptors has evolved to considerable complexity within the metazoans. The Frizzled-dependent signalling cascade comprises several branches, whose differential activation depends on specific Wnt ligands, Frizzled receptor isoforms and the cellular context. In Xenopus laevis embryos, the canonical β-catenin pathway contributes to the establishment of the dorsal–ventral axis. A different branch, referred to as the planar cell polarity pathway, is essential for cell polarization during elongation of the axial mesoderm by convergent extension. Here we demonstrate that a third branch of the cascade is independent of Dishevelled function and involves signalling through trimeric G proteins and protein kinase C (PKC). During gastrulation, Frizzled-7 (Fz7)-dependent PKC signalling controls cell-sorting behaviour in the mesoderm. Loss of zygotic Fz7 function results in the inability of involuted anterior mesoderm to separate from the ectoderm, which leads to severe gastrulation defects. This result provides a developmentally relevant in vivo function for the Fz/PKC pathway in vertebrates.

Cellular basis of Amphibian Gastrulation

Amphibian gastrulation is a complex integration of local cellular behavior to produce a supracellular system that, in turn, constrains and organizes the behavior of individual cells. Such behavior has fascinated and challenged embryologists for over a hundred years and has also perplexed some of them to the point of thinking it not reducible to part-processes. These thoughts were expressed by Walter Vogt (translated in Spemann, 1938), who did more than anyone to characterize the early morphogenesis of amphibians: It does not appear at all as if cells were walking in the sense, that single part movements were combining to form the movements of the masses; for even the most natural and plausible explanation by means of amoeboid moving of single cells fails utterly. We evidently have not the wandering of cells before us, but rather a passive obedience to a superior force.

Cell sorting in animal development: signalling and adhesive mechanisms in the formation of tissue boundaries

The organisation of the animal body into distinct tissues requires adhesive mechanisms that promote and maintain the physical segregation, the sorting, of different cell populations. Signals that control differential cell affinities across tissue boundaries have been identified, including Hedgehog, Notch, and EGF receptor signalling. Further, several examples demonstrate that cell sorting in vivo can be driven by Eph/ephrin signalling and by the differential expression of cadherins that modulate cell adhesion and motility.

Antero-posterior tissue polarity links mesoderm convergent extension to axial patterning

Remodelling its shape, or morphogenesis, is a fundamental property of living tissue. It underlies much of embryonic development and numerous pathologies. Convergent extension (CE) of the axial mesoderm of vertebrates is an intensively studied model for morphogenetic processes that rely on cell rearrangement. It involves the intercalation of polarized cells perpendicular to the antero-posterior (AP) axis, which narrows and lengthens the tissue. Several genes have been identified that regulate cell behaviour underlying CE in zebrafish and Xenopus. Many of these are homologues of genes that control epithelial planar cell polarity in Drosophila. However, elongation of axial mesoderm must be also coordinated with the pattern of AP tissue specification to generate a normal larval morphology. At present, the long-range control that orients CE with respect to embryonic axes is not understood. Here we show that the chordamesoderm of Xenopus possesses an intrinsic AP polarity that is necessary for CE, functions in parallel to Wnt/planar cell polarity signalling, and determines the direction of tissue elongation. The mechanism that establishes AP polarity involves graded activin-like signalling and directly links mesoderm AP patterning to CE.

Directional mesoderm cell migration in the Xenopus gastrula.

The movement of the dorsal mesoderm across the blastocoel roof of the Xenopus gastrula is examined. We show that different parts of the mesoderm which can be distinguished by their morphogenetic behavior in the embryo are all able to migrate independently on the inner surface of the blastocoel roof. The direction of mesoderm cell migration is determined by guidance cues in the extracellular matrix of the blastocoel roof and by an intrinsic tissue polarity of the mesoderm. The mesodermal polarity shows the same orientation as the external guidance cues and is strongly expressed in the more posterior mesoderm. The guidance cues of the extracellular matrix are recognized by all parts of the dorsal mesoderm and even by nonmesodermal cells from other regions of the embryo. The extracellular matrix consists of a network of fibronectin-containing fibrils. The adhesiveness of this matrix does not vary along the axis of mesoderm movement, excluding haptotaxis as a guidance mechanism in this system. However, an intact fibronectin fibril structure is necessary for directional mesoderm cell migration. When the assembly of fibronectin into fibrils is inhibited, mesoderm explants still migrate on the amorphous extracellular matrix, but no longer directionally. It is proposed that polarized extracellular matrix fibrils may normally guide the migrating mesoderm to its target region.

Guidance of mesoderm cell migration in the Xenopus gastrula requires PDGF signaling

In vertebrates, PDGFA and its receptor, PDGFRα, are expressed in the early embryo. Impairing their function causes an array of developmental defects, but the underlying target processes that are directly controlled by these factors are not well known. We show that in the Xenopus gastrula, PDGFA/PDGFRα signaling is required for the directional migration of mesodermal cells on the extracellular matrix of the blastocoel roof. Blocking PDGFRα function in the mesoderm does not inhibit migration per se, but results in movement that is randomized and no longer directed towards the animal pole. Likewise, compromising PDGFA function in the blastocoel roof substratum abolishes directionality of movement. Overexpression of wild-type PDGFA, or inhibition of PDGFA both lead to randomized migration, disorientation of polarized mesodermal cells, decreased movement towards the animal pole, and reduced head formation and axis elongation. This is consistent with an instructive role for PDGFA in the guidance of mesoderm migration.

EphrinB/EphB Signaling Controls Embryonic Germ Layer Separation by Contact-Induced Cell Detachment

Tissue boundary formation in the early vertebrate embryo involves cycles of cell attachment and detachment at the boundary, and cell contact-dependent signaling by membrane-bound EphB receptors and ephrinB ligands.

Epithelial coating controls mesenchymal shape change through tissue-positioning effects and reduction of surface-minimizing tension

Signalling between mesenchymal and epithelial cells has a profound influence on organ morphogenesis. However, less is known about the mechanical function of epithelial–mesenchymal interactions. Here, we describe two principal effects by which epithelia can regulate shape changes in mesenchymal cell aggregates. We propose that during formation of the embryonic body axis, the epithelial layer relieves surface minimizing tensions that would force cell aggregates into a spherical shape, and controls the serial arrangement of cell populations along the axis. The combined effects permit the tissue to deviate from a spherical form and to elongate.

Cadherin-dependent differential cell adhesion in Xenopus causes cell sorting in vitro, but not in the embryo

Adhesion differences between cell populations are in principle a source of strong morphogenetic forces promoting cell sorting, boundary formation and tissue positioning, and cadherins are main mediators of cell adhesion. However, a direct link between cadherin expression, differential adhesion and morphogenesis has not yet been determined for a specific process in vivo. To identify such a connection, we modulated the expression of C-cadherin in the Xenopus laevis gastrula, and combined this with direct measurements of cell adhesion-related parameters. Our results show that gastrulation is surprisingly tolerant of overall changes in adhesion. Also, as expected, experimentally generated, cadherin-based adhesion differences promote cell sorting in vitro. Importantly, however, such differences do not lead to the sorting of cells in the embryo, showing that differential adhesion is not sufficient to drive morphogenesis in this system. Compensatory recruitment of cadherin protein to contacts between cadherin-deprived and -overexpressing cells could contribute to the prevention of sorting in vivo.

Development of the lateral line system in Xenopus

The lateral line system of fishes and amphibians consists of numerous epidermal mechano-receptors which are distributed over the whole body surface. As in other amphibians, the lateral line system of Xenopus develops from epidermal placodes situated on the head region of the embryo. The dorsolateralis placodes form a rostro-caudal series of epidermal thickenings centered around the otic placode. In this series, placodes remaining within the epidermis and forming lateral line primordia alternate with lateral line ganglion forming placodes. Each lateral line primordium elongates and migrates within the epidermis along a well-defined pathway, leaving behind a row of small cell groups, the primary lateral line organs. As the ganglion which supplies a given row of organs and the corresponding lateral line primordium originate in spatial contiguity, and as the axons of the lateral line nerve grow out together with the migrating primordium, the lateral line neurones remain in contact with their target cells throughout development. After segregation of a primary organ from a migrating primordium, cell differentiation occurs. Receptor cells establish afferent and efferent synaptic contacts with axons from the lateral line nerve. Apically, a bundle of stereocilia and a single, microtubule-containing kinocilium protrude from the surface of a receptor cell into a jelly-like cupula, which extends into the surrounding fluid. Displacement of the cupula and the concomitant bending of the cilia stimulates the receptor cells. The cilia of a receptor cell are asymmetrically arranged, and this structural polarity is related to the directional sensitivity of the cells. Two types of receptor cells, with opposite orientations, are intermingled within each organ, giving the whole organ a bidirectional sensitivity. The number of lateral line organs is increased by the process of accessory organ formation, where primary organs grow and divide to produce secondary organs. In this way, existing rows of organs are extended. Moreover, single primary organs are transformed into elongate plaques of closely apposed organs. The lateral line system has reached its greatest extent at late larval stages. During metamorphosis, the number of organ plaques is reduced in some lines, and one line even disappears completely. Two large, myelinated afferent fibers innervate a whole organ plaque. They branch repeatedly to supply every organ of the plaque, and each fiber is thought to innervate only receptor cells of the same polarity.(ABSTRACT TRUNCATED AT 400 WORDS)