F-box protein FBXB-65 regulates anterograde transport of the kinesin-3 motor UNC-104 through a PTM near its cargo-binding PH domain.

Axonal transport in neurons is essential for cargo movement between the cell body and synapses. Caenorhabditis elegans UNC-104 and its homolog KIF1A are kinesin-3 motors that anterogradely transport precursors of synaptic vesicles (pre-SVs) and are degraded at synapses. However, in C. elegans, touch neuron-specific knockdown of the E1 ubiquitin-activating enzyme, uba-1, leads to UNC-104 accumulation at neuronal ends and synapses. Here, we performed an RNAi screen and identified that depletion of fbxb-65, which encodes an F-box protein, leads to UNC-104 accumulation at neuronal distal ends, and alters UNC-104 net anterograde movement and levels of UNC-104 on cargo without changing synaptic UNC-104 levels. Split fluorescence reconstitution showed that UNC-104 and FBXB-65 interact throughout the neuron. Our theoretical model suggests that UNC-104 might exhibit cooperative cargo binding that is regulated by FBXB-65. FBXB-65 regulates an unidentified post-translational modification (PTM) of UNC-104 in a region beside the cargo-binding PH domain. Both fbxb-65 and UNC-104, independently of FBXB-65, regulate axonal pre-SV distribution, transport of pre-SVs at branch points and organismal lifespan. FBXB-65 regulates a PTM of UNC-104 and the number of motors on the cargo surface, which can fine-tune cargo transport to the synapse.

Emergence of Spatial Scales and Macroscopic Tissue Dynamics in Active Epithelial Monolayers.

Migrating cells in tissues are often known to exhibit collective swirling movements. In this paper, we develop an active vertex model with polarity dynamics based on contact inhibition of locomotion (CIL). We show that under this dynamics, the cells form steady-state vortices in velocity, polarity, and cell stress with length scales that depend on polarity alignment rate (ζ), self-motility (v0), and cell-cell bond tension (λ). When the ratio λ/v0 becomes larger, the tissue reaches a near jamming state because of the inability of the cells to exchange their neighbors, and the length scale associated with tissue kinematics increases. A deeper examination of this jammed state provides insights into the mechanism of sustained swirl formation under CIL rule that is governed by the feedback between cell polarities and deformations. To gain additional understanding of how active forcing governed by CIL dynamics leads to large-scale tissue dynamics, we systematically coarse-grain cell stress, polarity, and motility and show that the tissue remains polar even on larger length scales. Overall, we explore the origin of swirling patterns during collective cell migration and obtain a connection between cell-level dynamics and large-scale cellular flow patterns observed in epithelial monolayers.

Percolation transition in phase-separating active fluid.

The motility-induced phase separation exhibited by active particles with repulsive interactions is well known. We show that the interaction softness of active particles destabilizes the highly ordered dense phase, leading to the formation of a porous cluster which spans the system. This soft limit can also be achieved if the particle motility is increased beyond a critical value, at which the system clearly exhibits all the characteristics of a standard percolation transition. We also show that in the athermal limit, active particles exhibit similar transitions even at low motility. With these additional new phases, the phase diagram of repulsive active particles is revealed to be richer than what was previously conceived.

Pseudocleavage furrows restrict plasma membrane-associated PH domain in syncytial Drosophila embryos.

Syncytial cells contain multiple nuclei and have local distribution and function of cellular components despite being synthesized in a common cytoplasm. The syncytial Drosophila blastoderm embryo shows reduced spread of organelle and plasma membrane-associated proteins between adjacent nucleo-cytoplasmic domains. Anchoring to the cytoarchitecture within a nucleo-cytoplasmic domain is likely to decrease the spread of molecules; however, its role in restricting this spread has not been assessed. In order to analyze the cellular mechanisms that regulate the rate of spread of plasma membrane-associated molecules in the syncytial Drosophila embryos, we express a pleckstrin homology (PH) domain in a localized manner at the anterior of the embryo by tagging it with the bicoid mRNA localization signal. Anteriorly expressed PH domain forms an exponential gradient in the anteroposterior axis with a longer length scale compared with Bicoid. Using a combination of experiments and theoretical modeling, we find that the characteristic distribution and length scale emerge due to plasma membrane sequestration and restriction within an energid. Loss of plasma membrane remodeling to form pseudocleavage furrows shows an enhanced spread of PH domain but not Bicoid. Modeling analysis suggests that the enhanced spread of the PH domain occurs due to the increased spread of the cytoplasmic population of the PH domain in pseudocleavage furrow mutants. Our analysis of cytoarchitecture interaction in regulating plasma membrane protein distribution and constraining its spread has implications on the mechanisms of spread of various molecules, such as morphogens in syncytial cells.

Dynamics and Stability of the Contractile Actomyosin Ring in the Cell.

Contraction of the cytokinetic ring during cell division leads to physical partitioning of a eukaryotic cell into two daughter cells. This involves flows of actin filaments and myosin motors in the growing membrane interface at the midplane of the dividing cell. Assuming boundary driven alignment of the actomyosin filaments at the inner edge of the interface, we explore how the resulting active stresses influence the flow. Using the continuum gel theory framework, we obtain exact axisymmetric solutions of the dynamical equations. These solutions are consistent with experimental observations on closure rate. Using these solutions, we perform linear stability analysis for the contracting ring under nonaxisymmetric deformations. Our analysis shows that few low wave number modes, which are unstable during onset of the constriction, later on become stable when the ring shrinks to smaller radii, which is a generic feature of actomyosin ring closure. Our theory also captures how the effective tension in the ring decreases with its radius, causing significant slowdown in the contraction process at later times.

Influence of interaction softness on phase separation of active particles.

Using a minimal model of active Brownian particles, we study the effect of a crucial parameter, namely the softness of the interparticle repulsion, on motility-induced phase separation. We show that an increase in particle softness reduces the ability of the system to phase separate and the system exhibits a delayed transition. After phase separation, the system state properties can be explained by a single relevant length scale, the effective interparticle distance. We estimate this length scale analytically and use it to rescale the state properties at dense phase for systems with different interaction softness. Using this length scale, we provide a scaling relation for the time taken to phase separate which shows a high sensitivity to the interaction softness.

Cyto-architecture constrains the spread of photoactivated tubulin in the syncytial Drosophila embryo.

Drosophila embryogenesis begins with nuclear division in a common cytoplasm forming a syncytial cell. Morphogen gradient molecules spread across nucleo-cytoplasmic domains to pattern the body axis of the syncytial embryo. The diffusion of molecules across the syncytial nucleo-cytoplasmic domains is potentially constrained by association with the components of cellular architecture. However, the extent of restriction has not been examined. Here we use photoactivation (PA) to generate a source of cytoplasmic or cytoskeletal molecules in order to monitor the kinetics of their spread in the syncytial Drosophila embryo. Photoactivated PA-GFP and PA-GFP-Tubulin generated within a fixed anterior area diffused along the antero-posterior axis. These molecules were enriched in the cortical cytoplasm above the yolk-filled center, suggesting that the cortical cytoplasm is phase separated from the yolk-filled center. The length scales of diffusion were extracted using exponential fits under steady state assumptions. PA-GFP spread a greater distance as compared to PA-GFP-Tubulin. Both molecules were more restricted when generated in the center of the embryo. The length scale of spread for PA-GFP-Tubulin increased in mutant embryos containing short plasma membrane furrows and a disrupted tubulin cytoskeleton. PA-GFP spread was unaffected by cyto-architecture perturbation. Taken together, these data show that PA-GFP-Tubulin spread is restricted by its incorporation in the microtubule network and intact plasma membrane furrows. This photoactivation based analysis of protein spread allows for interpretation of the dependence of gradient formation on syncytial cyto-architecture.

Non-Gaussian subdiffusion of single-molecule tracers in a hydrated polymer network.

Single molecule tracking experiments inside a hydrated polymer network have shown that the tracer motion is subdiffusive due to the viscoelastic environment inside the gel-like network. This property can be related to the negative autocorrelation of the instantaneous displacements at short times. Although the displacements of the individual tracers exhibit Gaussian statistics, the displacement distribution of all the trajectories combined from different spatial locations of the polymer network exhibits a non-Gaussian distribution. Here, we analyze many individual tracer trajectories to show that the central portion of the non-Gaussian distribution can be well approximated by an exponential distribution that spreads sublinearly with time. We explain all these features seen in the experiment by a generalized Langevin model for an overdamped particle with algebraically decaying correlations. We show that the degree of non-Gaussianity can change with the extent of heterogeneity, which is controlled in our model by the experimentally observed distributions of the motion parameters.

Capture of a diffusive prey by multiple predators in confined space.

The first passage search of a diffusing target (prey) by multiple searchers (predators) in confinement is an important problem in the stochastic process literature. While the analogous problem in open space has been studied in some detail, a systematic study in confined space is still lacking. In this paper, we study the first passage times for this problem in one, two, and three dimensions. Due to confinement, the survival probability of the target takes a form ∼e^{-t/τ} at large times t. The characteristic capture timescale τ associated with the rare capture events are rather challenging to measure. We use a computational algorithm that allows us to estimate τ with high accuracy. We study in detail the behavior of τ as a function of the system parameters, namely, the number of searchers N, the relative diffusivity r of the target with respect to the searcher, and the system size. We find that τ deviates from the ∼1/N scaling seen in the case of a static target, and this deviation varies continuously with r and the spatial dimensions.

Energy transfer-mediated white light emission from Nile red-doped 9,10-diphenylanthracene nanoaggregates upon excitation with near UV light.

The low cost, ease of preparation, colour tunability and wide application range garnered huge research interest on organic light emitting diode materials (OLED). The development of white light-emitting organic diode materials is mostly targeted for this. Anthracene derivatives have recently emerged as low-cost and efficient blue light-emitting diodes. However, developing efficient organic diode materials that cover the entire visible spectrum is very challenging. Herein, we demonstrated that Nile red (NR)-doped 9,10-diphenylanthracene (DPA) nanoaggregates provided strong white light emission upon excitation with near UV light. The dual emissions of the DPA nanoaggregates covering the blue and green regions were exploited and combined with the controlled red emission of the properly doped NR dye to cover the full visible spectrum, rendering white light emission with a quantum yield of >0.4. The fluorescence spectra of the DPA nanoaggregates doped with NR at various concentrations were monitored and their CIE coordinates were followed to evaluate the proper doping ratio for equal-energy white-light emission. Concurrent time-resolved emission studies provided mechanistic insights into the energy transfer from the exciton and excimer states of DPA to NR. It was revealed that the energy transfer from the singlet excitonic state of DPA followed the diffusion-assisted resonance energy transfer (RET) model. On the other hand, the excimer state showed negligible diffusion and energy transfer from this state found to follow the single-step Förster resonance energy transfer mechanism. The observation of efficient white light emission in the doped DPA nanoaggregates was proposed to have prospective applications in OLED devices, given the fact that triplet excitons may be exploited for emission through the efficient triplet-triplet annihilation contribution to fluorescence enhancement.

Ultrafast Conformational Relaxation Dynamics in Anthryl-9-benzothiazole: Dynamic Planarization Driven Delocalization and Protonation-Induced Twisting Dynamics.

Conformational motion in the excited state of fluorophores critically governs their photophysical properties. Unveiling controlling parameters of photoinduced molecular motion in organic dyes is essential for optimization of light-triggered processes. Herein, we present ultrafast dynamics of conformational relaxation controlled photophysical properties of anthryl-9-benzthiazole (AnBT). The title compound is a bichromophore, consists of anthracene (AN) and benzothiazole (BT) units connected by a single bond, and exists in out of plane ground state conformation (dihedral angle of about 65?). Vibronic resolved structured absorption feature of anthracene localized excitation is lost in first excited singlet state displaying large Stokes-shifted fluorescence, characteristics of delocalized state involving both AN and BT unit. Ultrafast transient absorption spectroscopic studies revealed evolution of anthracene-localized Franck?Condon state to a delocalized excited state in a few picosecond time scale depending on solvent viscosity. A planarized motion of AN and BT units is proposed to be involved in excited state relaxation. However, protonation of BT unit is shown to induce significant intramolecular charge transfer facilitated by ultrafast torsional relaxation promoting nonradiative deactivation. Thus, depending upon protonation state, AnBT is shown to undergo either ultrafast planarized motion facilitating delocalized emissive excited state or perpendicular torsion rendering nonemissive twisted intramolecular charge transfer state. Experimental results were corroborated by quantum chemical calculation.

Interplay of exciton-excimer dynamics in 9,10-diphenylanthracene nanoaggregates and thin films revealed by time-resolved spectroscopic studies.

Anthracene and its derivatives are organic semiconducting materials that have prospective applications in organic solar cells and organic light emitting diodes. A thorough understanding of the photophysics and exciton dynamics is imperative for the effective utilization of these materials in optoelectronics. Herein, we have presented the exciton dynamics of a highly emissive anthracene derivative, namely, 9,10-diphenylanthracene (DPA) in nanoaggregate, thin film and crystalline forms. In contrast to the strong blue molecular emission in solution, DPA in the solid form exhibits emission from both the exciton and excimer state, covering the blue and green spectral region. In the well-ordered crystalline state, excimer emission dominates, while in the nanoaggregate form, the relative contribution of the excimer state decreases with the decrease in the size of the nanoparticle. In crystals, thin films and larger size nanoaggregates, favourable intermolecular orbital overlap by the adjacent phenyl substituents of DPA in the dominant α-phase packing allows for the fast relaxation of the exciton to excimer state in a sub-nanosecond timescale, as measured by time-resolved emission studies. Prior to trapping in the excimer state, the diffusion of singlet excitons has been revealed via the exciton-exciton annihilation kinetics measured by the excitation fluence dependent ultrafast emission decay. The singlet exciton diffusion coefficient in the DPA nanoaggregate is noted to be almost an order of magnitude slower than that in the anthracene nanoaggregate previously reported. The near perpendicular orientation of the two phenyl rings at the 9 and 10 position of anthracene causes an elongation in the intermolecular distance of the anthracene cores, which impedes the exciton diffusion rate. However, the ordered molecular packing of the DPA molecules in the thin film and crystal facilitates significantly faster singlet exciton diffusion.

First passage of a particle in a potential under stochastic resetting: A vanishing transition of optimal resetting rate.

First passage in a stochastic process may be influenced by the presence of an external confining potential, as well as "stochastic resetting" in which the process is repeatedly reset back to its initial position. Here, we study the interplay between these two strategies, for a diffusing particle in a one-dimensional trapping potential V(x), being randomly reset at a constant rate r. Stochastic resetting has been of great interest as it is known to provide an "optimal rate" (r_{*}) at which the mean first passage time is a minimum. On the other hand, an attractive potential also assists in the first capture process. Interestingly, we find that for a sufficiently strong external potential, the advantageous optimal resetting rate vanishes (i.e., r_{*}→0). We derive a condition for this optimal resetting rate vanishing transition, which is continuous. We study this problem for various functional forms of V(x), some analytically, and the rest numerically. We find that the optimal rate r_{*} vanishes with a deviation from the critical strength of the potential as a power law with an exponent ß which appears to be universal.

Intrinsic stochastic resonance via set-point variation.

In the present paper, the possibility of invoking stochastic resonance (SR, periodic and aperiodic) by regulating the operating value of an appropriate parameter is explored. The operating values of these parameters are defined as the set point of the system throughout the present paper. Brusselator, a mathematical model [I. Prigogine and R. Lefever, J. Chem. Phys. 48, 1695 (1968)JCPSA60021-960610.1063/1.1668896] of nonlinear chemical reactions, is used for this purpose. We consider the effect of intrinsic noise in the Brusselator due to the Markovian nature of the chemical reactions. The stochastic time evolution is studied using the Gillespie algorithm [D. T. Gillespie, J. Comput. Phys. 22, 403 (1976)JCTPAH0021-999110.1016/0021-9991(76)90041-3], which is an exact stochastic simulation algorithm. We analyze the dependence of the resonance point on both the strength of the intrinsic noise as well as the distance from the bifurcation point. Subsequently, the phenomena of SR is explored using both periodic and aperiodic stimulus. It was found that, for a given system size, in both cases, SR is achieved by variation of the set point. Set-point variation can be achieved by regulating either the source concentration or the rate constants. Resonance is observed in both cases. However, this resonance occurs at different values of the set point, even with a fixed system size. This is clearly seen in the set-point versus system-size plane, where the resonance line has different slopes for the two scenarios. Our semianalytic treatment points to the fact that for a given system size intrinsic noise is affected differently for different methods involving the variation of the set point. This is explained by writing the corresponding chemical Langevin equation and comparing the various intrinsic noise sources.

Protonation-induced ultrafast torsional dynamics in 9-anthrylbenzimidazole: a pH activated molecular rotor.

We report the photophysical properties and excited state dynamics of 9-anthrylbenzimidazole (ANBI) which exhibits protonation-induced molecular rotor properties. In contrast to the highly emissive behavior of neutral ANBI, protonation of the benzimidazole group of ANBI induces efficient nonradiative deactivation by ultrafast torsional motion around the bond connecting the anthracene and benzimidazole units, as revealed by ultrafast transient absorption and fluorescence spectroscopy. Contrary to viscosity-independent fluorescence of neutral dyes, protonated ANBI is shown to display linear variation of emission yield and lifetime with solvent viscosity. The protonation-induced molecular rotor properties in the studied system are shown to be driven by enhanced charge transfer and are corroborated by quantum chemical calculations. Potential application as a microviscosity sensor of acidic regions in a heterogeneous environment by these proton-activated molecular rotor properties of ANBI is discussed.

Solvent sensitive intramolecular charge transfer dynamics in the excited states of 4-N,N-dimethylamino-4'-nitrobiphenyl.

Organic molecules substituted with the nitro group show efficient nonlinear optical (NLO) properties, which are a consequence of the strong intramolecular charge transfer (ICT) character of the molecules because of the strong electron withdrawing nature of the nitro group and rapid responsiveness because of highly movable π-electrons. Dynamics of the ICT process in the excited states of a push-pull biphenyl derivative, namely, 4-N,N-dimethylamino-4'-nitrobiphenyl (DNBP), an efficient NLO material, has been investigated using ultrafast transient absorption spectroscopy. The experimental results have been corroborated with DFT and TDDFT calculations. In solvents of large polarity, e.g. acetonitrile, the ultrafast ICT process of DNBP is associated with the barrierless twisting of the N,N-dimethylaniline (DMA) group with respect to the nitrobenzene moiety to populate the twisted ICT (or TICT) state, and the rate of this process is solely governed by the viscosity of the medium. In solvents of moderate polarity, e.g. ethyl acetate, the rate of the twisting process is significantly slowed down and the LE and TICT states remain in equilibrium because of a low energy barrier for interconversion between these two states. By further lowering the polarity of the solvent, e.g. in dioxane, the twisting process is completely retarded. In nonpolar solvents, e.g. cyclohexane, a reverse twisting motion towards the planar geometry (i.e. the PICT process) has been evident in the excited state dynamics. In this solvent, the S1 state undergoes an ultrafast intersystem crossing to the triplet state because of its close proximity with the T2 state.

Interplay of cell dynamics and epithelial tension during morphogenesis of the Drosophila pupal wing.

How tissue shape emerges from the collective mechanical properties and behavior of individual cells is not understood. We combine experiment and theory to study this problem in the developing wing epithelium of Drosophila. At pupal stages, the wing-hinge contraction contributes to anisotropic tissue flows that reshape the wing blade. Here, we quantitatively account for this wing-blade shape change on the basis of cell divisions, cell rearrangements and cell shape changes. We show that cells both generate and respond to epithelial stresses during this process, and that the nature of this interplay specifies the pattern of junctional network remodeling that changes wing shape. We show that patterned constraints exerted on the tissue by the extracellular matrix are key to force the tissue into the right shape. We present a continuum mechanical model that quantitatively describes the relationship between epithelial stresses and cell dynamics, and how their interplay reshapes the wing.

Cell-fibronectin interactions propel vertebrate trunk elongation via tissue mechanics.

During embryonic development and tissue homeostasis, cells produce and remodel the extracellular matrix (ECM). The ECM maintains tissue integrity and can serve as a substrate for cell migration. Integrin α5 (Itgα5) and αV (ItgαV) are the α subunits of the integrins most responsible for both cell adhesion to the ECM protein fibronectin (FN) and FN matrix fibrillogenesis. We perform a systems-level analysis of cell motion in the zebrafish tail bud during trunk elongation in the presence and absence of normal cell-FN interactions. Itgα5 and ItgαV have well-described roles in cell migration in vitro. However, we find that concomitant loss of itgα5 and itgαV leads to a trunk elongation defect without substantive alteration of cell migration. Tissue-specific transgenic rescue experiments suggest that the FN matrix on the surface of the paraxial mesoderm is required for body elongation via its role in defining tissue mechanics and intertissue adhesion.

Plasticization of poly(vinylpyrrolidone) thin films under ambient humidity: insight from single-molecule tracer diffusion dynamics.

Studies on diffusion dynamics of single molecules (SMs) have been useful in revealing inhomogeneity of polymer thin films near and above the glass-transition temperature (T(g)). However, despite several applications of polymer thin films where exposure to solvent (or vapor) is common, the effect of absorbed solvent molecules on local morphology and rigidity of polymer matrices is yet to be explored in detail. High-T(g) hydrophilic polymers such as poly(vinylpyrrolidone) (PVP) are used as pharmaceutical coatings for drug release in aqueous medium, as they readily absorb moisture, which results in effective lowering of the T(g) and thereby leads to plasticization. The effect of moisture absorption on swelling and softening of PVP thin films was investigated by visualizing the diffusion dynamics of rhodamine 6G (Rh6G) tracer molecules at various ambient relative humidities (RH). Wide-field epifluorescence microscopy, in conjunction with high-resolution SM tracking, was used to monitor the spatiotemporal evolution of individual tracers under varied moisture contents of the matrix. In the absence of atmospheric moisture, Rh6G molecules in dry PVP films are translationally inactive, suggestive of rigid local environments. Under low moisture contents (RH 30-50%), translational mobility remains arrested but rotational motion is augmented, indicating slight swelling of the polymer network which marks the onset of plasticization. The translational mobility of Rh6G was found to be triggered only at a threshold ambient RH, beyond which a large proportion of tracers exhibit extensive diffusion dynamics. Interestingly, SM tracking data at higher moisture contents of the film (RH ≥ 60%) reveal that the distributions of dynamic parameters (such as diffusivity) are remarkably broad, spanning several orders of magnitude. Furthermore, Rh6G molecules display a wide variety of translational motion even at a fixed ambient RH, clearly pointing out the extremely inhomogeneous environment of plasticized PVP network. Intriguingly, it is observed that a majority of tracers undergo anomalous subdiffusion even under high moisture contents of the matrix. Analyses of SM trajectories using velocity autocorrelation function reveal that subdiffusive behaviors of Rh6G are likely to originate from fractional Brownian motion, a signature of tracer dynamics in viscoelastic medium.

Regulated tissue fluidity steers zebrafish body elongation.

The tailbud is the posterior leading edge of the growing vertebrate embryo and consists of motile progenitors of the axial skeleton, musculature and spinal cord. We measure the 3D cell flow field of the zebrafish tailbud and identify changes in tissue fluidity revealed by reductions in the coherence of cell motion without alteration of cell velocities. We find a directed posterior flow wherein the polarization between individual cell motion is high, reflecting ordered collective migration. At the posterior tip of the tailbud, this flow makes sharp bilateral turns facilitated by extensive cell mixing due to increased directional variability of individual cell motions. Inhibition of Wnt or Fgf signaling or cadherin 2 function reduces the coherence of the flow but has different consequences for trunk and tail extension. Modeling and additional data analyses suggest that the balance between the coherence and rate of cell flow determines whether body elongation is linear or whether congestion forms within the flow and the body axis becomes contorted.