Integrin-ECM Interactions Regulate Cadherin-Dependent Cell Adhesion and Are Required for Convergent Extension in Xenopus

AbstractBackground: Convergence extension movements are conserved tissue rearrangements implicated in multiple morphogenetic events. While many of the cell behaviors involved in convergent extension are known, the molecular interactions required for this process remain elusive. However, past evidence suggests that regulation of cell adhesion molecule function is a key step in the progression of these behaviors.Results: Antibody blocking of fibronectin (FN) adhesion or dominant-negative inhibition of integrin β1 function alters cadherin-mediated cell adhesion, promotes cell-sorting behaviors in reaggregation assays, and inhibits medial-lateral cell intercalation and axial extension in gastrulating embryos and explants. Embryo explants were used to demonstrate that normal integrin signaling is required for morphogenetic movements within defined regions but not for cell fate specification. The binding of soluble RGD-containing fragments of fibronectin to integrins promotes the reintegration of dissociated single cells into intact tissues. The changes in adhesion observed are independent of cadherin or integrin expression levels.Conclusions: We conclude that integrin modulation of cadherin adhesion influences cell intercalation behaviors within boundaries defined by extracellular matrix. We propose that this represents a fundamental mechanism promoting localized cell rearrangements throughout development

Integrin α5β1 and Fibronectin Regulate Polarized Cell Protrusions Required for Xenopus Convergence and Extension

SummaryBackgroundIntegrin recognition of fibronectin is required for normal gastrulation including the mediolateral cell intercalation behaviors that drive convergent extension and the elongation of the frog dorsal axis; however, the cellular and molecular mechanisms involved are unclear.ResultsWe report that depletion of fibronectin with antisense morpholinos blocks both convergent extension and mediolateral protrusive behaviors in explant preparations. Both chronic depletion of fibronectin and acute disruptions of integrin α5β1 binding to fibronectin increases the frequency and randomizes the orientation of polarized cellular protrusions, suggesting that integrin-fibronectin interactions normally repress frequent random protrusions in favor of fewer mediolaterally oriented ones. In the absence of integrin α5β1 binding to fibronectin, convergence movements still occur but result in convergent thickening instead of convergent extension.ConclusionsThese findings support a role for integrin signaling in regulating the protrusive activity that drives axial extension. We hypothesize that the planar spatial arrangement of the fibrillar fibronectin matrix, which delineates tissue compartments within the embryo, is critical for promoting productive oriented protrusions in intercalating cells

Peptide and peptide-carbon nanotube hydrogels as scaffolds for tissue & 3D tumor engineering

The final publication is available at Elsevier via http://dx.doi.org/10.1016/j.actbio.2017.12.012 © 2018. This manuscript version is made available under the CC-BY-NC-ND 4.0 license https://creativecommons.org/licenses/by-nc-nd/4.0/The use of hybrid self-assembling peptide (EFK8)-carbon nanotube (SWNT) hydrogels for tissue engineering and in vitro 3D cancer spheroid formation is reported. These hybrid hydrogels are shown to enhance the attachment, spreading, proliferation and movement of NIH-3T3 cells relative to that observed using EFK8-only hydrogels. After five days, ∼30% more cells are counted when the hydrogel contains SWNTs. Also, 3D encapsulation of these cells when injected in hydrogels does not adversely affect their behavior. Compressive modulus measurements and microscopic examination suggest that SWNTs have this beneficial effect by providing sites for cell anchorage, spreading and movement rather than by increasing hydrogel stiffness. This shows that the cells have a particular interaction with SWNTs not shared with EFK8 nanofibers despite a similar morphology. The effect of EFK8 and EFK8-SWNT hydrogels on A549 lung cancer cell behavior is also investigated. Increasing stiffness of EFK8-only hydrogels from about 44 Pa to 104 Pa promotes a change in A549 morphology from spheroidal to a stretched one similar to migratory phenotype. EFK8-SWNT hydrogels also promote a stretched morphology, but at lower stiffness. These results are discussed in terms of the roles of both microenvironment stiffness and cell-scaffold adhesion in cancer cell invasion. Overall, this study demonstrates that applications of peptide hydrogels in vitro can be expanded by incorporating SWNTs into their structure which further provides insight into cell-biomaterial interactions. Statement of significance For the first time we used hybrid self-assembling peptide-carbon nanotube hybrid hydrogels (that we have recently introduced briefly in the “Carbon” journal in 2014) for tissue engineering and 3D tumor engineering. We showed the potential of these hybrid hydrogels to enhance the efficiency of the peptide hydrogels for tissue engineering application in terms of cell behavior (cell attachment, spreading and migration). This opens up new rooms for the peptide hydrogels and can expand their applications. Also our system (peptide and peptide-CNT hydrogels) was used for cancer cell spheroid formation showing the effect of both tumor microenvironment stiffness and cell-scaffold adhesion on cancer cell invasion. This was only possible based on the presence of CNTs in the hydrogel while the stiffness kept constant. Finally it should be noted that these hybrid hydrogels expand applications of peptide hydrogels through enhancing their capabilities and/or adding new properties to them.Natural Sciences and Engineering Research Council of Canada (NSERC)Canada Foundation for Innovation (CFI)Canada Research Chairs (CRC) progra

Integrin α5β1 Function Is Regulated by XGIPC/kermit2 Mediated Endocytosis during Xenopus laevis Gastrulation

During Xenopus gastrulation α5β1 integrin function is modulated in a temporally and spatially restricted manner, however, the regulatory mechanisms behind this regulation remain uncharacterized. Here we report that XGIPC/kermit2 binds to the cytoplasmic domain of the α5 subunit and regulates the activity of α5β1 integrin. The interaction of kermit2 with α5β1 is essential for fibronectin (FN) matrix assembly during the early stages of gastrulation. We further demonstrate that kermit2 regulates α5β1 integrin endocytosis downstream of activin signaling. Inhibition of kermit2 function impairs cell migration but not adhesion to FN substrates indicating that integrin recycling is essential for mesoderm cell migration. Furthermore, we find that the α5β1 integrin is colocalized with kermit2 and Rab 21 in embryonic and XTC cells. These data support a model where region specific mesoderm induction acts through kermit2 to regulate the temporally and spatially restricted changes in adhesive properties of the α5β1 integrin through receptor endocytosis

Search for new phenomena in events with a photon and missing transverse momentum in pp collisions at root s=13 TeV with the ATLAS detector

Results of a search for new phenomena in events with an energetic photon and large missing transverse momentum with the ATLAS experiment at the Large Hadron Collider are reported. The data were collected in proton--proton collisions at a centre-of-mass energy of 13 TeV and correspond to an integrated luminosity of 3.2 $\rm fb^{-1}$. The observed data are in agreement with the Standard Model expectations. Exclusion limits are presented in models of new phenomena including pair production of dark matter candidates or large extra spatial dimensions. In a simplified model of dark matter and an axial-vector mediator, the search excludes mediator masses of up to 710 GeV for dark matter candidate masses up to 150 GeV. In an effective theory of dark matter production, values of the suppression scale $M_*$ up to 570 GeV are excluded and the effect of truncation for various coupling values is reported. For the ADD large extra spatial dimension model the search places more stringent limits than earlier searches in the same event topology, excluding $M_{\rm D}$ up to about 2.3 (2.8) TeV for two (six) additional spatial dimensions; the limits are reduced by 20--40% depending on the number of additional spatial dimensions when applying a truncation procedure

Stress-induced synthesis of pyruvate kinase in xenopus laevis embryos

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