Rab5-mediated endocytosis of activin is not required for gene activation or long-range signalling in Xenopus.
ABSTRACT Morphogen gradients provide positional cues for cell fate specification and tissue patterning during embryonic development. One important aspect of morphogen function, the mechanism by which long-range signalling occurs, is still poorly understood. In Xenopus, members of the TGF-beta family such as the nodal-related proteins and activin act as morphogens to induce mesoderm and endoderm. In an effort to understand the mechanisms and dynamics of morphogen gradient formation, we have used fluorescently labelled activin to study ligand distribution and Smad2/Smad4 bimolecular fluorescence complementation (BiFC) to analyse, in a quantitative manner, the cellular response to induction. Our results indicate that labelled activin travels exclusively through the extracellular space and that its range is influenced by numbers of type II activin receptors on responding cells. Inhibition of endocytosis, by means of a dominant-negative form of Rab5, blocks internalisation of labelled activin, but does not affect the ability of cells to respond to activin and does not significantly influence signalling range. Together, our data indicate that long-range signalling in the early Xenopus embryo, in contrast to some other developmental systems, occurs through extracellular movement of ligand. Signalling range is not regulated by endocytosis, but is influenced by numbers of cognate receptors on the surfaces of responding cells.
Conference Paper: Using autoregressive spectral analysis to investigate weaning index[Show abstract] [Hide abstract]
ABSTRACT: Mechanical ventilation is widely used in lifesaving. But it is associated with numerous complications. Patients having weaning failure are about 7% to 19% and are subject to high mortality. To investigate this problem, we use a digital signal process approach. We record breathing data before and after weaning. The patients' weaning success and weaning failure are also recorded. The autoregressive power spectral density method is used to quantify the breathing data and investigate its rhythm changes. From spectral analysis, the power ratio of the first two peaks in the low frequency regions is calculated. We observed that patients with weaning failure have this ratio less than 1. This ratio may be a good weaning index.Engineering in Medicine and Biology, 2002. 24th Annual Conference and the Annual Fall Meeting of the Biomedical Engineering Society EMBS/BMES Conference, 2002. Proceedings of the Second Joint; 02/2002
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ABSTRACT: Morphogens are substances that establish a graded distribution and elicit distinct cellular responses in a dose-dependent manner. They function to provide individual cells within a field with positional information, which is interpreted to give rise to spatial patterns. Morphogens can consist of intracellular factors that set up a concentration gradient by diffusion in the cytoplasm. More commonly, morphogens comprise secreted proteins that form an extracellular gradient across a field of cells. Experimental studies and computational analyses have provided support for a number of diverse strategies by which extracellular morphogen gradients are formed. These include free diffusion in the extracellular space, restricted diffusion aided by interactions with heparan sulfate proteoglycans, transport on lipid-containing carriers or transport aided by soluble binding partners. More specialized modes of transport have also been postulated such as transcytosis, in which repeated rounds of secretion, endocytosis, and intracellular trafficking move morphogens through cells rather than around them, or cytonemes, which consist of filopodial extensions from signal-receiving cells that are hypothesized to reach out to morphogen-sending cells. Once the gradient has formed, cells must distinguish small differences in morphogen concentration and store this information even after the gradient has dissipated. This is often achieved by translating ligand concentration into a proportional increase in numbers of activated cell surface receptors that are internalized and continue to signal from endosomal compartments. Ultimately, this leads to activation of one or a few transcription factors that transduce this information into qualitatively distinct gene responses inside the nucleus. WIREs Dev Biol 2012, 1:3-15. doi: 10.1002/wdev.2 For further resources related to this article, please visit the WIREs website.Wiley interdisciplinary reviews. Developmental biology. 01/2012; 1(1):3-15.
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ABSTRACT: The yeast two-hybrid system pioneered the field of in vivo protein-protein interaction methods and undisputedly gave rise to a palette of ingenious techniques that are constantly pushing further the limits of the original method. Sensitivity and selectivity have improved because of various technical tricks and experimental designs. Here we present an exhaustive overview of the genetic approaches available to study in vivo binary protein interactions, based on two-hybrid and protein fragment complementation assays. These methods have been engineered and employed successfully in microorganisms such as Saccharomyces cerevisiae and Escherichia coli, but also in higher eukaryotes. From single binary pairwise interactions to whole-genome interactome mapping, the self-reassembly concept has been employed widely. Innovative studies report the use of proteins such as ubiquitin, dihydrofolate reductase, and adenylate cyclase as reconstituted reporters. Protein fragment complementation assays have extended the possibilities in protein-protein interaction studies, with technologies that enable spatial and temporal analyses of protein complexes. In addition, one-hybrid and three-hybrid systems have broadened the types of interactions that can be studied and the findings that can be obtained. Applications of these technologies are discussed, together with the advantages and limitations of the available assays.Microbiology and molecular biology reviews: MMBR 06/2012; 76(2):331-82. · 12.59 Impact Factor