A Cryptic Frizzled Module in Cell Surface Collagen 18 Inhibits Wnt/β−Catenin Signaling

Collagens contain cryptic polypeptide modules that regulate major cell functions, such as cell proliferation or death. Collagen XVIII (C18) exists as three amino terminal end variants with specific amino terminal polypeptide modules. We investigated the function of the variant 3 of C18 (V3C18) containing a frizzled module (FZC18), which carries structural identity with the extracellular cysteine-rich domain of the frizzled receptors. We show that V3C18 is a cell surface heparan sulfate proteoglycan, its topology being mediated by the FZC18 module. V3C18 mRNA was expressed at low levels in 21 normal adult human tissues. Its expression was up-regulated in fibrogenesis and in small well-differentiated liver tumors, but decreased in advanced human liver cancers. Low FZC18 immunostaining in liver cancer nodules correlated with markers of high Wnt/β−catenin activity. V3C18 (Mr = 170 kD) was proteolytically processed into a cell surface FZC18-containing 50 kD glycoprotein precursor that bound Wnt3a in vitro through FZC18 and suppressed Wnt3a-induced stabilization of β−catenin. Ectopic expression of either FZC18 (35 kD) or its 50 kD precursor inhibited Wnt/β−catenin signaling in colorectal and liver cancer cell lines, thus downregulating major cell cycle checkpoint gatekeepers cyclin D1 and c-myc and reducing tumor cell growth. By contrast, full-length V3C18 was unable to inhibit Wnt signaling. In summary, we identified a cell-surface signaling pathway whereby FZC18 inhibits Wnt/β−catenin signaling. The signal, encrypted within cell-surface C18, is released by enzymatic processing as an active frizzled cysteine-rich domain (CRD) that reduces cancer cell growth. Thus, extracellular matrix controls Wnt signaling through a collagen-embedded CRD behaving as a cell-surface sensor of proteolysis, conveying feedback cues to control cancer cell fate

The GABAergic system: a possible component of estrogenic feedback on gonadotropin secretion in rainbow trout (Oncorhynchus mykiss)

In teleost fish, GTH secretion is controlled by a large number of neuroendocrine factors at the central level and steroid feedback represents an efficient process to synchronize the activity of all the systems involved along the brain-pituitary-gonad axis. Estrogen effects are mediated by specific nuclear receptors that act as transcription factors to regulate the expression of target genes. In order to understand the neuroendocrine mechanisms involved in the estrogen feedback on GTH secretion, we need, as a first step, to know the nature of target cells at the pituitary and central levels. In rainbow trout, some estrogen receptors expressing cells are identified but the nature of a large number of them remains unknown. In this paper, we explain our strategy to identify the central target of estrogen using both in situ hybridization and immunohistochemistry. We focused our attention on GABA neurons of which the distribution, in some central areas, exactly overlaps with that of estrogen receptors expressing cells

Caspase-2, a Novel Lipid Sensor under the Control of Sterol Regulatory Element Binding Protein 2

Caspases play important roles in apoptotic cell death and in some other functions, such as cytokine maturation, inflammation, or differentiation. We show here that the 5′-flanking region of the human CASP-2 gene contains three functional response elements for sterol regulatory element binding proteins (SREBPs), proteins that mediate the transcriptional activation of genes involved in cholesterol, triacylglycerol, and fatty acid synthesis. Exposure of several human cell lines to statins, lipid-lowering drugs that drive SREBP proteolytic activation, induced the CASP-2 gene to an extent similar to that for known targets of SREBP proteins. Adenoviral vector-mediated transfer of active SREBP-2 also induced expression of the CASP-2 gene and the caspase-2 protein and increased the cholesterol and triacylglycerol cellular content. These rises in lipids were strongly impaired following small interfering RNA-mediated silencing of the CASP-2 gene. Taken together, our results identify the human CASP-2 gene as a member of the SREBP-responsive gene battery that senses lipid levels in cells and raise the possibility that caspase-2 participates in the control of cholesterol and triacylglycerol levels

Size Segregation in a Fluid-like or Gel-like Suspension Settling under Gravity or in a Centrifuge

We investigate size segregation effects in a bidisperse concentrated suspension when slowly settling under gravity or when submitted to a centrifugal field. Experiments are carried out with PMMA spheres of two different mean diameters (190 and 25 μm) suspended in a hydrophobic index-matched fluid. Spatial repartitions of both small and large spheres and velocity fluctuations of particles are measured using fluorescently labeled PMMA spheres and a particle-image-velocimetry method. Large particles behave as hard spheres in purely hydrodynamic interactions, while small spheres interact through weakly attractive forces. For a small amount of small spheres among large ones, the suspension remains fluid during settling and the organization of the velocity field of particles into finite-sized structures also called "blobs" promotes size segregation. A larger proportion of weakly attractive small spheres in the bidisperse suspension causes a considerable slowdown of the settling process under gravity and the occurrence of a large-scale collective behavior together with a loss of size segregation. When centrifuging the gel-like bidisperse suspension, a shear-induced melting of the particle network induces a spectacular segregation of species. As a consequence, aging tests of soft yielding materials using centrifugation methods are not representative of the shelf-life stability of the products. A tentative model based on the competition between viscous stresses acting upon particles and adhesive stresses gives a correct estimate of the critical stationary acceleration for the destabilization of the particle network and the onset of size segregation in a gel-like suspension