β-Adrenergic Inhibition of Contractility in L6 Skeletal Muscle Cells

The β-adrenoceptors (β-ARs) control many cellular processes. Here, we show that β-ARs inhibit calcium depletion-induced cell contractility and subsequent cell detachment of L6 skeletal muscle cells. The mechanism underlying the cell detachment inhibition was studied by using a quantitative cell detachment assay. We demonstrate that cell detachment induced by depletion of extracellular calcium is due to myosin- and ROCK-dependent contractility. The β-AR inhibition of L6 skeletal muscle cell detachment was shown to be mediated by the β2-AR and increased cAMP but was surprisingly not dependent on the classical downstream effectors PKA or Epac, nor was it dependent on PKG, PI3K or PKC. However, inhibition of potassium channels blocks the β2-AR mediated effects. Furthermore, activation of potassium channels fully mimicked the results of β2-AR activation. In conclusion, we present a novel finding that β2-AR signaling inhibits contractility and thus cell detachment in L6 skeletal muscle cells by a cAMP and potassium channel dependent mechanism

Protein kinase C activation disrupts epithelial apical junctions via ROCK-II dependent stimulation of actomyosin contractility

<p>Abstract</p> <p>Background</p> <p>Disruption of epithelial cell-cell adhesions represents an early and important stage in tumor metastasis. This process can be modeled <it>in vitro </it>by exposing cells to chemical tumor promoters, phorbol esters and octylindolactam-V (OI-V), known to activate protein kinase C (PKC). However, molecular events mediating PKC-dependent disruption of epithelial cell-cell contact remain poorly understood. In the present study we investigate mechanisms by which PKC activation induces disassembly of tight junctions (TJs) and adherens junctions (AJs) in a model pancreatic epithelium.</p> <p>Results</p> <p>Exposure of HPAF-II human pancreatic adenocarcinoma cell monolayers to either OI-V or 12-O-tetradecanoylphorbol-13-acetate caused rapid disruption and internalization of AJs and TJs. Activity of classical PKC isoenzymes was responsible for the loss of cell-cell contacts which was accompanied by cell rounding, phosphorylation and relocalization of the F-actin motor nonmuscle myosin (NM) II. The OI-V-induced disruption of AJs and TJs was prevented by either pharmacological inhibition of NM II with blebbistatin or by siRNA-mediated downregulation of NM IIA. Furthermore, AJ/TJ disassembly was attenuated by inhibition of Rho-associated kinase (ROCK) II, but was insensitive to blockage of MLCK, calmodulin, ERK1/2, caspases and RhoA GTPase.</p> <p>Conclusion</p> <p>Our data suggest that stimulation of PKC disrupts epithelial apical junctions via ROCK-II dependent activation of NM II, which increases contractility of perijunctional actin filaments. This mechanism is likely to be important for cancer cell dissociation and tumor metastasis.</p

Manifestation of the final states in completely momentum resolved coincidence spectroscopy RID G-7348-2011

We present the general method of analysis of the six-dimensional data measured in the momentum resolved e,2e reaction on the solid surfaces. The basis of the method is a normalization of the experimental density distribution on the model experimental function that describes the volume of available final states for this reaction in the momentum space. The density distributions of correlated electron pairs as functions of various variables are considered for the e,2e scattering on the Fe(110) surface. (C) 2002 Elsevier Science B.V. All rights reserved

The use of the time-energy dispersion in an electron energy analyzer RID G-7348-2011

The time-energy dispersion (TED) characteristics of a spherical electrostatic mirror are used to design an unusual electron energy analyzer. It is shown that TED of a retarding spherical field in an electron mirror configuration is positive and it increases with the electron kinetic energy. This is due to an increasing penetration of electrons with high kinetic energy in the retarding field and provides the basis for developing a new type of time-of-flight spectrometer (TOF). Two limiting cases for the geometry of the proposed spectrometer are considered and for both of them the energy resolution of the TOF device depends on the combined time resolution of electron detector and readout electronics. In the case of a 10-cm inner sphere radius (22 cm outer sphere radius) for the electrostatic mirror, the spectrometer is expected to have an energy resolution of about 0.5 eV/ns and an acceptance solid angle of about 2.2 srad for electrons of 75 eV kinetic energy. A position sensitive detector allows retrieving the energy and the emission angle on the basis of the measured time-of-flight and detection point position. In the second case the inner sphere radius is 6 cm and the outer spherical segment has a radius of 120 cm. The energy resolution is expected to be about 160 meV/ns at the electron kinetic energy of 1 keV and the acceptance solid angle is 0.1 srad. In both cases changing the retarding potential can easily change the energy range within which the TOF analysis is performed. (C) 2003 Elsevier B.V. All rights reserved

Spatially restricted activation of RhoA signalling at epithelial junctions by p114RhoGEF drives junction formation and morphogenesis

Signalling by the GTPase RhoA, a key regulator of epithelial cell behaviour, can stimulate opposing processes: RhoA can promote junction formation and apical constriction, and reduce adhesion and cell spreading. Molecular mechanisms are thus required that ensure spatially restricted and process-specific RhoA activation. For many fundamental processes, including assembly of the epithelial junctional complex, such mechanisms are still unknown. Here we show that p114RhoGEF is a junction-associated protein that drives RhoA signalling at the junctional complex and regulates tight-junction assembly and epithelial morphogenesis. p114RhoGEF is required for RhoA activation at cell-cell junctions, and its depletion stimulates non-junctional Rho signalling and induction of myosin phosphorylation along the basal domain. Depletion of GEF-H1, a RhoA activator inhibited by junctional recruitment, does not reduce junction-associated RhoA activation. p114RhoGEF associates with a complex containing myosin II, Rock II and the junctional adaptor cingulin, indicating that p114RhoGEF is a component of a junction-associated Rho signalling module that drives spatially restricted activation of RhoA to regulate junction formation and epithelial morphogenesis