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Synapses are semi-membraneless, protein-dense, sub-micron chemical reaction compartments responsible for signal processing in each and every neuron. Proper formation and dynamic responses to stimulations of synapses, both during development and in adult, are fundamental to functions of mammalian brains, although the molecular basis governing formation and modulation of compartmentalized synaptic assemblies is unclear. Here, we used a biochemical reconstitution approach to show that, both in solution and on supported membrane bilayers, multivalent interaction networks formed by major excitatory postsynaptic density (PSD) scaffold proteins led to formation of PSD-like assemblies via phase separation. The reconstituted PSD-like assemblies can cluster receptors, selectively concentrate enzymes, promote actin bundle formation, and expel inhibitory postsynaptic proteins. Additionally, the condensed phase PSD assemblies have features that are distinct from those in homogeneous solutions and fit for synaptic functions. Thus, we have built a molecular platform for understanding how neuronal synapses are formed and dynamically regulated.
Assuntos
Neurogênese , Plasticidade Neuronal , Densidade Pós-Sináptica , Sinapses/fisiologia , Animais , Encéfalo/fisiologia , Proteína 4 Homóloga a Disks-Large/fisiologia , Hipocampo/fisiologia , Luz , Camundongos , Microscopia Confocal , Neurônios/fisiologia , Espalhamento de Radiação , Transdução de Sinais , Transmissão SinápticaRESUMO
Phase segregation is ubiquitously observed in immiscible mixtures, such as oil and water, in which the mixing entropy is overcome by the segregation enthalpy1-3. In monodispersed colloidal systems, however, the colloidal-colloidal interactions are usually non-specific and short-ranged, which leads to negligible segregation enthalpy4. The recently developed photoactive colloidal particles show long-range phoretic interactions, which can be readily tuned with incident light, suggesting an ideal model for studying phase behaviour and structure evolution kinetics5,6. In this work, we design a simple spectral selective active colloidal system, in which TiO2 colloidal species were coded with spectral distinctive dyes to form a photochromic colloidal swarm. In this system, the particle-particle interactions can be programmed by combining incident light with various wavelengths and intensities to enable controllable colloidal gelation and segregation. Furthermore, by mixing the cyan, magenta and yellow colloids, a dynamic photochromic colloidal swarm is formulated. On illumination of coloured light, the colloidal swarm adapts the appearance of incident light due to layered phase segregation, presenting a facile approach towards coloured electronic paper and self-powered optical camouflage.
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We report direct atomic force microscopy measurements of pinning-depinning dynamics of a circular moving contact line (CL) over the rough surface of a micron-sized vertical hanging glass fiber, which intersects a liquid-air interface. The measured capillary force acting on the CL exhibits sawtoothlike fluctuations, with a linear accumulation of force of slope k (stick) followed by a sharp release of force δf, which is proportional to the CL slip length. From a thorough analysis of a large volume of the stick-slip events, we find that the local maximal force F_{c} needed for CL depinning follows the extreme value statistics and the measured δf follows the avalanche dynamics with a power law distribution in good agreement with the Alessandro-Beatrice-Bertotti-Montorsi (ABBM) model. The experiment provides an accurate statistical description of the CL dynamics at mesoscale, which has important implications to a common class of problems involving stick-slip motion in a random defect or roughness landscape.
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Direct measurement and control of the dynamic wetting properties of a lipid-coated water-air interface over a wide range of surface tension variations have many important applications. However, the wetting dynamics of the interface near its partial-to-complete wetting transition has not been fully understood. Here, we report a systematic study of the wetting dynamics of a lipid-coated water-air interface around a thin glass fiber of diameter 1-5 µm and length 100-300 µm. The glass fiber is glued onto the front end of a rectangular cantilever to form a "long-needle" atomic-force-microscope probe. Three surface modifications are applied to the glass fiber to change its wetting properties from hydrophilic to hydrophobic. A monolayer of phospholipid dipalmitoylphosphatidylcholine (DPPC) is deposited on the water-air interface in a homemade Langmuir-Blodgett trough, and the surface tension γL of the DPPC-coated water-air interface is varied in the range of 2.5 ⲠγL â² 72 mN/m. From the measured hysteresis loop of the capillary force for the three coated fiber surfaces with varying γL, we observe a sharp transition from partial to complete wetting when γL is reduced to a critical value (γL)c. The obtained values of (γL)c are 27 ± 1 mN/m for a DPPC-coated fiber surface and 23 ± 1 mN/m for an trichloro(1H,1H,2H,2H-perfluorooctyl) silane (FTS)-coated surface. Below (γL)c, the contact angle θ0 of the liquid interface is found to be zero for both hydrophobic fiber surfaces and the corresponding spreading parameter S becomes positive. For the FTS-coated fiber surface, the height of capillary rise exhibits a jump when γL is reduced to (γL)c, which indicates that a rapidly advancing liquid film is formed on the fiber surface when the partial-to-complete wetting transition takes place. Our experiment thus establishes a quantitative method by which many other liquid interfaces coated with polymers, surfactants, and biomolecules (such as proteins and lipids) may be characterized dynamically.
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Inspired by the experimental and numerical findings, we study the dynamic instabilities of two coupled nonlinear delay differential equations that are used to describe the coherent oscillations between the top and bottom boundary layers in turbulent Rayleigh-Bénard convection. By introducing two sensitivity parameters for the instabilities of the top and bottom boundary layers, we find three different types of solutions, namely in-phase single-period oscillations, multi-period oscillations and chaos. The chaos solution contains rare but large amplitude fluctuations. The statistical properties of these fluctuations are consistent with those observed in the experiment for the massive eruption of thermal plumes, which causes random reversals of the large-scale circulation in turbulent Rayleigh-Bénard convection. Our study thus provides new insights into the origin of rare massive eruptions and sudden changes of large-scale flow pattern that are often observed in convection systems of geophysical and astrophysical scales.
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We report direct atomic-force-microscope measurements of capillary force hysteresis (CFH) of a circular contact line (CL) formed on a long glass fiber, which is coated with a thin layer of soft polymer film and intersects a water-air interface. The measured CFH shows a distinct overshoot for the depinning of a static CL, and the overshoot amplitude grows logarithmically with both the hold time τ and fiber speed V. A unified model based on the slow growth of a wetting ridge and force-assisted barrier crossing is developed to explain the observed time (or state) and speed (or rate) dependent CL depinning dynamics over an aging soft surface. The experimental findings have important implications to a common class of problems involving depinning dynamics in a defect or roughness landscape, such as friction of solid interfaces.
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Most organs contain interconnected tubular tissues that are one-cell-thick, polarized epithelial monolayers enclosing a fluid-filled lumen. Such tissue organization plays crucial roles in developmental and normal physiology, and the proper functioning of these tissues depends on their regulation by complex biochemical perturbations and equally important, but poorly understood, mechanical perturbations. In this study, by combining micropatterning techniques and atomic force microscopy, we developed a simple in vitro experimental platform for characterizing the mechanical properties of the MDCK II cyst, the simplest model of lumen-enclosing epithelial monolayers. By using this platform, we estimated the elasticity of the cyst monolayer and showed that the presence of a luminal space influences cyst mechanics substantially, which could be attributed to polarization and tissue-level coordination. More interestingly, the results from force-relaxation experiments showed that the cysts also displayed tissue-level poroelastic characteristics that differed slightly from those of single cells. Our study provides the first quantitative findings, to our knowledge, on the tissue-level mechanics of well-polarized epithelial cysts and offers new insights into the interplay between cyst mechanics and cyst physiology. Moreover, our simple platform is a potentially useful tool for enhancing the current understanding of cyst mechanics in health and disease.
Assuntos
Engenharia Celular , Elasticidade , Células Epiteliais/citologia , Microscopia de Força Atômica , Microtecnologia , Animais , Fenômenos Biomecânicos , Cães , Células Madin Darby de Rim CaninoRESUMO
We report the noncontact measurement of the viscoelastic property of polymer thin films in a liquid medium using frequency-modulation atomic force microscopy with a newly developed long-needle probe. The probe contains a long vertical glass fiber with one end adhered to a cantilever beam and the other end with a sharp tip placed near the liquid-film interface. The nanoscale flow generated by the resonant oscillation of the needle tip provides a precise hydrodynamic force acting on the soft surface of the thin film. By accurately measuring the mechanical response of the thin film, we obtain the elastic and loss moduli of the thin film using the linear response theory of elastohydrodynamics. The experiment verifies the theory and demonstrates its applications. The technique can be used to accurately measure the viscoelastic property of soft surfaces, such as those made of polymers, nanobubbles, live cells, and tissues.
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A two-layer colloidal system is developed for the study of diffusion over a quenched two-dimensional random potential. A mixture of bidisperse silica spheres is used to form a randomly packed colloidal monolayer on the bottom substrate. The corrugated surface of the bottom colloidal monolayer provides a gravitational potential field for the dilute diffusing particles in the top layer. The population probability histogram P(x,y) of the diffusing particles is obtained to fully characterize the random potential landscape U(x,y) via the Boltzmann distribution. The dynamical properties of the top diffusing particles, such as their mean square displacement (MSD), histogram of the escape time, and long-time self-diffusion coefficient, are simultaneously measured from the particle trajectories. A quantitative relationship between the long-time diffusion coefficient and the random potential is obtained, which is in good agreement with the theoretical prediction. The measured MSD reveals a wide region of subdiffusion resulting from the structural disorders. The crossover from subdiffusion to normal diffusion is explained by the Lorentz model for tracer diffusion through a heterogeneous space filled with a set of randomly distributed obstacles.
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We report a systematic study of colloidal diffusion over a substrate with quasicrystalline-patterned holes. Silica spheres of diameter comparable to the hole diameter diffuse over the patterned substrate and experience a gravitational potential U(x, y). Using optical microscopy, we track the particle trajectories and find two distinct states: a trapped state when the particles are inside the holes and a free-diffusion state when they are on the flat surface outside the holes. The potential U(x, y) and dynamic properties of the diffusing particle, such as its mean dwell time, mean square displacement, and long-time diffusion coefficient DL, are measured simultaneously. The measured DL is in good agreement with the prediction of two theoretical models proposed for diffusion over a quasicrystal lattice. The experiment demonstrates the applications of this newly constructed potential landscape.
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We report on direct atomic-force-microscope measurements of capillary force hysteresis (CFH) and relaxation of a circular moving contact line (CL) formed on a long micron-sized hydrophobic fiber intersecting a water-air interface. The measured CFH and CL relaxation show a strong asymmetric speed dependence in the advancing and receding directions. A unified model based on force-assisted barrier crossing is utilized to find the underlying energy barrier Eb and size λ associated with the defects on the fiber surface. The experiment demonstrates that the pinning (relaxation) and depinning dynamics of the CL can be described by a common microscopic framework, and the advancing and receding CLs are influenced by two different sets of relatively wetting and nonwetting defects on the fiber surface.
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A tilted two-layer colloidal system is constructed for the study of force-assisted barrier-crossing dynamics over a periodic potential. The periodic potential is provided by the bottom layer colloidal spheres forming a fixed crystalline pattern on a glass substrate. The corrugated surface of the bottom colloidal crystal provides a gravitational potential field for the top layer diffusing particles. By tilting the sample at an angle θ with respect to the vertical (gravity) direction, a tangential component of the gravitational force F is applied to the diffusing particles. The measured mean drift velocity v(F, Eb) and diffusion coefficient D(F, Eb) of the particles as a function of F and energy barrier height Eb agree well with the exact results of the one-dimensional drift velocity (R. L. Stratonovich, Radiotekh. Elektron, 1958, 3, 497) and diffusion coefficient (P. Reimann, et al., Phys. Rev. Lett., 2001, 87, 010602 and P. Reimann, et al., Phys. Rev. E, 2002, 65, 031104). Based on these exact results, we show analytically and verify experimentally that there exists a scaling region, in which v(F, Eb) and D(F, Eb) both scale as ν'(F)exp[-E(F)/kBT], where the Arrhenius pre-factor ν'(F) and effective barrier height E(F) are both modified by F. The experiment demonstrates the applications of this model system in evaluating different scaling forms of ν'(F) and E(F) and their accuracy, in order to extract useful information about the external potential, such as the intrinsic barrier height Eb.
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Background: The development of effective inhibitors that can inhibit amyloid ß (Aß) peptides aggregation and promote neurite outgrowth is crucial for the possible treatment of Alzheimer's disease (AD). Lobaria (Schreb.) Hoffm., a traditional Chinese medicine used in Himalaya region for inflammatory diseases, contains depsides/depsidones (DEPs) such as gyrophoric acid, norstictic acid, and stictic acid known for their anti-cancer and anti-inflammation properties. Methods: Lobaria extracts were analyzed using HPLC to identify DEPs and establish standards. The inhibitory effects of Lobaria on Aß42 fibrillization and depolymerization were assessed using various approaches with biophysical and cellular methods. The neuroprotective activity of Lobaria extracts and its DEPs aganist Aß-mediated cytotoxicity was also evaluated. Results: Norstictic and stictic acid were found in the water extract, while norstictic, stictic, and gyrophoric acid were detected in the ethanol extract of Lobaria. Both extracts, and their DEPs effectively inhibited Aß42 fibrillation and disaggregate mature Aß42 fibrils. Notably, the ethanol extract showed superior inhibitory effect compared to the water extract, with gyrophoric acid being the most effective DEPs. Additionally, herbal extract-treated Aß42 aggregation species significantly protected neuronal cells from Aß42-induced cell damage and promoted neurite outgrowth. Conclusion: This study is the first to investigate the effect of Lobaria on Aß42 and neuronal cell in AD. Given that Lobaria is commonly used in ethnic medicine and food with good safety records, our findings propose that Lobaria extracts and DEPs have potential as neuroprotective and therapeutic agents for AD patients.
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Using the techniques of optical microscopy and particle tracking, we measure the pair correlation function and Brownian diffusion in monolayers of strongly interacting colloidal particles suspended at or near three different interfaces and test the universal scaling law of the normalized diffusion coefficient, D[over Ë]≃e(αΔS), as a function of the excess entropy ΔS for a wide range of particle concentrations. It is found that the universal scaling law with α=1 holds well for highly charged polystyrene spheres suspended at an air-water interface, where the strong electrostatic interactions play a dominant role. For monolayer suspensions of hard-sphere-like particles, where hydrodynamic interactions become important, deviations from the universal scaling law are observed. The experiment indicates that the hydrodynamic corrections could be incorporated into the universal scaling law of diffusion with an exponent α<1.
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Using the techniques of optical microscopy and particle tracking, we measure the correlated diffusion in a monolayer of uniform silica spheres dispersed at a water-air interface. It is found that the correlated motion of the interfacial particles can be well described by two universal response functions, the normalized longitudinal and transverse diffusion coefficients D(â¥)(r/r0) and D(â¥)(r/r0), where r is the interparticle distance and r0=a(λS/a)(3/2) is a new scaling length, which depends on both the Saffman length λS and particle radius a. The obtained response functions characterize the crossover behavior of the colloidal monolayers from the subphase-dominated three-dimensional hydrodynamics at low surface coverage to the monolayer-dominated 2D hydrodynamics at high concentrations. The surface viscosity ηs(2) of the colloidal monolayer obtained by two-particle rheology compares well with the one-particle measurements.
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We report a direct measurement of the friction coefficient of a fluctuating (and slipping) contact line using a thin vertical glass fiber of diameter d with one end glued onto a cantilever beam and the other end touching a liquid-air interface. By measuring the broadening of the resonant peak of the cantilever system with varying liquid viscosity η, we find the friction coefficient of the contact line has a universal form, ξ(c)≃0.8πdη, independent of the liquid-solid contact angle. The obtained scaling law is further supported by the numerical simulation based on the phase field model under the generalized Navier boundary conditions.
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Friction between two rough solid surfaces often involves local stick-slip events occurring at different locations of the contact interface. If the apparent contact area is large, multiple local slips may take place simultaneously and the total frictional force is a sum of the pinning forces imposed by many asperities on the interface. Here, we report a systematic study of stick-slip friction over a mesoscale contact area using a hanging-beam lateral atomic-force-microscope, which is capable of resolving frictional force fluctuations generated by individual slip events and measuring their statistical properties at the single-slip resolution. The measured probability density functions (PDFs) of the slip length δxs, the maximal force Fc needed to trigger the local slips, and the local force gradient [Formula: see text] of the asperity-induced pinning force field provide a comprehensive statistical description of stick-slip friction that is often associated with the avalanche dynamics at a critical state. In particular, the measured PDF of δxs obeys a power law distribution and the power-law exponent is explained by a new theoretical model for the under-damped spring-block motion under a Brownian-correlated pinning force field. This model provides a long-sought physical mechanism for the avalanche dynamics in stick-slip friction at mesoscale.
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We report a systematic study of the dynamics of self-propelled particles (SPPs) over a one-dimensional periodic potential landscape U_{0}(x), which is fabricated on a microgroove-patterned polydimethylsiloxane (PDMS) substrate. From the measured nonequilibrium probability density function P(x;F_{0}) of the SPPs, we find that the escape dynamics of the slow rotating SPPs across the potential landscape can be described by an effective potential U_{eff}(x;F_{0}), once the self-propulsion force F_{0} is included into the potential under the fixed angle approximation. This work demonstrates that the parallel microgrooves provide a versatile platform for a quantitative understanding of the interplay among the self-propulsion force F_{0}, spatial confinement by U_{0}(x), and thermal noise, as well as its effects on activity-assisted escape dynamics and transport of the SPPs.
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A well-defined oscillation is observed in the power spectrum of several fluctuating signals in turbulent Rayleigh-Bénard convection occurring in a thin vertical disk filled with water. The experiment reveals that the coherent oscillations are produced by periodic emission of thermal plumes, which gives rise to periodic pulses of forcing, resulting in a pulsed large-scale circulation in the thin cell. The experimental results agree well with the theoretical predictions made from two coupled nonlinear delayed equations.
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We studied the two-dimensional freezing transitions in monolayers of microgel colloidal spheres with short-ranged repulsions in video-microscopy experiments, and monolayers of hard disks, and Yukawa particles in simulations. These systems share two common features at the freezing points: (1) the bimodal distribution profile of the local orientational order parameter; (2) the two-body excess entropy, s(2), reaches -4.5±0.5 k(B). Both features are robust and sensitive to the freezing points, so that they can potentially serve as empirical freezing criteria in two dimensions. Compared with the conventional freezing criteria, the first feature has no finite-size ambiguities and can be resolved adequately with much less statistics; and the second feature can be directly measured in macroscopic experiments without the need for microscopic information.