E-cadherin homophilic ligation directly signals through Rac and phosphatidylinositol 3-kinase to regulate adhesive contacts

Classical cadherins mediate cell recognition and cohesion in many tissues of the body. It is increasingly apparent that dynamic cadherin contacts play key roles during morphogenesis and that a range of cell signals are activated as cells form contacts with one another. It has been difficult, however, to determine whether these signals represent direct downstream consequences of cadherin ligation or are juxtacrine signals that are activated when cadherin adhesion brings cell surfaces together but are not direct downstream targets of cadherin signaling. In this study, we used a functional cadherin ligand (hE/Fc) to directly test whether E-cadherin ligation regulates phosphatidylinositol 3-kinase (PI 3-kinase) and Rac signaling. We report that homophilic cadherin ligation recruits Rae to nascent adhesive contacts and specifically stimulates Rae signaling. Adhesion to hE/Fc also recruits PI 3-kinase to the cadherin complex, leading to the production of phosphatidylinositol 3,4,5-trisphosphate in nascent cadherin contacts. Rae activation involved an early phase, which was PI 3-kinase-independent, and a later amplification phase, which was inhibited by wortmannin. PI 3-kinase and Rae activity were necessary for productive adhesive contacts to form following initial homophilic ligation. We conclude that E-cadherin is a cellular receptor that is activated upon homophilic ligation to signal through PI 3-kinase and Rae. We propose that a key function of these cadherin-activated signals is to control adhesive contacts, probably via regulation of the actin cytoskeleton, which ultimately serves to mediate adhesive cell-cell recognition.

Charting the course of ovarian development in vertebrates

The decision of the embryonic gonad to differentiate as either a testis or an ovary is a critical step in vertebrate development. The molecular basis of this decision has been the focus of much study, particularly over the past decade. Here we contrast the knowledge of early gonadal development and the switch to testis differentiation with the lack of molecular understanding of ovarian development at early stages. We review current knowledge regarding mechanisms of ovarian morphogenesis and propose a model for the hierarchical control of development of the fetal ovary, incorporating the few genes already known to be important and several signals or factors that are hypothesised to exist in the early ovary.

Exogenous Slit2 does not affect ureteric branching or nephron formation during kidney development

In an attempt to elucidate the role of Slit2 invertebrate kidney development, the effect of adding exogenous human Slit2 protein (hSlit2) to developing murine metanephric kidney explants was examined. To confirm the activity of the recombinant Slit2 protein, neurons from 8 day old chick sympathetic nerve chain dorsal root ganglia were cultured with hSlit2 protein, which induced significant neurite branching and outgrowth. Using kidney explants as a model system, metanephric development in the presence of hSlit2 protein was examined. Addition of hSlit2 up to a final concentration of 1 mug/ml had no detectable effect on the formation of nephrons or on branching morphogenesis of the ureteric tree after 2 or 4 days in culture, as assessed via immunofluorescence for the markers WT1 and calbindin 28K respectively. Similarly, maturation of the nephrogenic mesenchyme occurred in a phenotypically normal fashion. In situ analysis of the Slit receptors, Robot and Robot, the vasculogenic markers VEGFA and Flk-1, and the stromal cell marker BF2 displayed no difference in comparison to controls.

The embryonic development of the bodywall and nervous system of the cestode flatworm Hymenolepis diminuta

Cestodes (tapeworms) are a derived, parasitic clade of the phylum Platyhelminthes (flatworms). The cestode body wall represents an adaptation to its endoparasitic lifestyle. The epidermis forms a nonciliated syncytium, and both muscular and nervous system are reduced. Morphological differences between cestodes and free-living flatworms become apparent already during early embryogenesis. Cestodes have a complex life cycle that begins with an infectious larva, called the oncosphere. In regard to cell number, cestode oncospheres are among the simplest multicellular organisms, containing in the order of 50-100 cells. As part of our continuing effort to analyze embryonic development in flatworms, we describe here the staining pattern obtained with acTub in embryos and larvae of the cestode Hymenolepis diminuta and, briefly, the monogenean Neoheterocotyle rhinobatidis. In addition, we labeled the embryonic musculature of Hymenolepis with phalloidin. In Hymenolepis embryos, two different cell types that we interpret as neurons and epidermal gland cells express acTub. There exist only two neurons that develop close to the midline at the anterior pole of the embryo. The axons of these two neurons project posteriorly into the center of the oncosphere, where they innervate the complex of muscles that is attached to the booklets. In addition to neurons, acTub labels a small and invariant set of epidermal gland cells that develop at superficial positions, anteriorly adjacent to the neurons, in the dorsal midline, and around the posteriorly located hooklets. During late stages of embryogenesis they spread and form a complete covering of the embryo. We discuss these data in the broader context of platyhelminth embryology.

Distribution of a nematocyst-bearing sponge in relation to potential coral donors

Haliclona sp. 628 (Demospongiae, Haplosclerida, Chalinidae), a sponge found on the reef slope below 5 in depth on the Great Barrier Reef, has two unusual characteristics. It contains a symbiotic dinoflagellate, Symbiodinium sp., similar in structure to the dinoflagellate found within Acropora nobilis (S. microadriaticum), and it contains coral nematocysts randomly distributed between the ectosome and endosome and usually undischarged in intact sponge tissue. Given the unusual occurrence of nematocysts in Haliclona sp. 628, the focus of this study was to determine the distribution of this species of sponge on the reef slope at Heron Island Reef in relation to the distribution of potential coral donors. A combination of line and belt transects was used to estimate the abundance of Halielona sp. 628 and a co-occurring congener, Haliclona sp. 1031, which does not contain nematocysts, at three widely separated sites on the reef slope at Heron Island Reef. The abundance of different types of substratum (sand, sand-covered coral rubble, dead A. nobilis, live A. nobilis, other live coral, and other dead coral) along the transects and the substratum to which each sponge colony was attached were also recorded. Despite the predominance of live A. nobilis and sand-covered rubble at all sites, between 30 and 55% of Haliclona sp. 628 colonies were attached to dead A. nobilis which comprised less than 8% of the available substratum along any transect. In contrast, Haliclona sp. 1031 was found significantly more frequently on other dead corals and less frequently on live A. nobilis than would be expected based on the availability of the different substrata in the sites. Potential explanations to account for the distribution of Haliclona sp. 628 in relation to potential coral donors are discussed.

Linking physiological and architectural models of cotton

Despite the strong influence of plant architecture on crop yield, most crop models either ignore it or deal with it in a very rudimentary way. This paper demonstrates the feasibility of linking a model that simulates the morphogenesis and resultant architecture of individual cotton plants with a crop model that simulates the effects of environmental factors on critical physiological processes and resulting yield in cotton. First the varietal parameters of the models were made concordant. Then routines were developed to allocate the flower buds produced each day by the crop model amongst the potential positions generated by the architectural model. This allocation is done according to a set of heuristic rules. The final weight of individual bolls and the shedding of buds and fruit caused by water, N, and C stresses are processed in a similar manner. Observations of the positions of harvestable fruits, both within and between plants, made under a variety of agronomic conditions that had resulted in a broad range of plant architectures were compared to those predicted by the model with the same environmental inputs. As illustrated by comparisons of plant maps, the linked models performed reasonably well, though performance of the fruiting point allocation and shedding algorithms could probably be improved by further analysis of the spatial relationships of retained fruit. (C) 2002 Elsevier Science Ltd. All rights reserved.

Characterization of E-cadherin endocytosis in isolated MCF-7 and Chinese hamster ovary cells - The initial fate of unbound E-cadherin

The endocytosis of E-cadherin has recently emerged as an important determinant of cadherin function with the potential to participate in remodeling adhesive contacts. In this study we focused on the initial fate of E-cadherin when it predominantly exists free on the cell surface prior to adhesive binding or incorporation into junctions. Surface-labeling techniques were used to define the endocytic itinerary of E-cadherin in MCF-7 cells and in Chinese hamster ovary cells stably expressing human E-cadherin. We found that in this experimental system E-cadherin entered a transferrin-negative compartment before transport to the early endosomal compartment, where it merged with classical clathrin-mediated uptake pathways. E-cadherin endocytosis was inhibited by mutant dynamin, but not by an Eps15 mutant that effectively blocked transferrin internalization. Furthermore, sustained signaling by the ARF6 GTPase appeared to trap endocytosed E-cadherin in large peripheral structures. We conclude that in isolated cells unbound E-cadherin on the cell surface is predominantly endocytosed by a clathrin-independent pathway resembling macropinocytotic internalization, which then fuses with the early endosomal system. Taken with earlier reports, this suggests the possibility that multiple pathways exist for E-cadherin entry into cells that are likely to reflect cell context and regulation.