ABSTRACT
Biomolecular condensates enable cell compartmentalization by acting as membraneless organelles1. How cells control the interactions of condensates with other cellular structures such as membranes to drive morphological transitions remains poorly understood. We discovered that formation of a tight-junction belt, which is essential for sealing epithelial tissues, is driven by a wetting phenomenon that promotes the growth of a condensed ZO-1 layer2 around the apical membrane interface. Using temporal proximity proteomics in combination with imaging and thermodynamic theory, we found that the polarity protein PATJ mediates a transition of ZO-1 into a condensed surface layer that elongates around the apical interface. In line with the experimental observations, our theory of condensate growth shows that the speed of elongation depends on the binding affinity of ZO-1 to the apical interface and is constant. Here, using PATJ mutations, we show that ZO-1 interface binding is necessary and sufficient for tight-junction belt formation. Our results demonstrate how cells exploit the collective biophysical properties of protein condensates at membrane interfaces to shape mesoscale structures.
Subject(s)
Biomolecular Condensates , Cell Membrane , Tight Junctions , Wettability , Animals , Dogs , Humans , Biomolecular Condensates/metabolism , Biomolecular Condensates/chemistry , Cell Compartmentation , Cell Membrane/metabolism , Cell Membrane/chemistry , Epithelium , HEK293 Cells , Madin Darby Canine Kidney Cells , Mutation , Protein Binding , Thermodynamics , Tight Junction Proteins/metabolism , Tight Junctions/metabolism , Tight Junctions/chemistry , Zonula Occludens-1 Protein/genetics , Zonula Occludens-1 Protein/metabolism , ProteomicsABSTRACT
How complex 3D tissue shape emerges during animal development remains an important open question in biology and biophysics. Here, we discover a mechanism for 3D epithelial shape change based on active, in-plane cellular events that is analogous to inanimate "shape programmable" materials, which undergo blueprinted 3D shape transformations from in-plane gradients of spontaneous strains. We study eversion of the Drosophila wing disc pouch, when the epithelium transforms from a dome into a curved fold, quantifying 3D tissue shape changes and mapping spatial patterns of cellular behaviors on the evolving geometry using cellular topology. Using a physical model inspired by shape programming, we find that active cell rearrangements are the major contributor to pouch eversion and validate this conclusion using a knockdown of MyoVI, which reduces rearrangements and disrupts morphogenesis. This work shows that shape programming is a mechanism for animal tissue morphogenesis and suggests that patterns in nature could present design strategies for shape-programmable materials.
Subject(s)
Morphogenesis , Wings, Animal , Animals , Wings, Animal/growth & development , Drosophila Proteins/metabolism , Drosophila Proteins/genetics , Drosophila melanogaster/growth & development , Drosophila , Models, Biological , Imaginal Discs/metabolism , Imaginal Discs/growth & developmentABSTRACT
A fundamental question about biomolecular condensates is how distinct condensates can emerge from the interplay of different components. Here we present a minimal model of droplet differentiation where phase separated droplets demix into two types with different chemical modifications triggered by enzymatic reactions. We use numerical solutions to Cahn-Hilliard equations with chemical reactions and an effective droplet model to reveal the switchlike behavior. Our work shows how condensate identities in cells could result from competing enzymatic actions.
ABSTRACT
Hertwig's rule states that cells divide along their longest axis, usually driven by forces acting on the mitotic spindle. Here, we show that in contrast to this rule, microtubule-based pulling forces in early Caenorhabditis elegans embryos align the spindle with the short axis of the cell. We combine theory with experiments to reveal that in order to correct this misalignment, inward forces generated by the constricting cytokinetic ring rotate the entire cell until the spindle is aligned with the cell's long axis. Experiments with slightly compressed mouse zygotes indicate that this cytokinetic ring-driven mechanism of ensuring Hertwig's rule is general for cells capable of rotating inside a confining shell, a scenario that applies to early cell divisions of many systems.
Subject(s)
Caenorhabditis elegans , Spindle Apparatus , Animals , Caenorhabditis elegans/embryology , Mice , Spindle Apparatus/metabolism , Microtubules/metabolism , Cytokinesis/physiology , Rotation , Zygote/metabolism , Zygote/cytology , Zygote/growth & development , Embryo, Nonmammalian/cytology , Embryonic Development/physiology , Models, BiologicalABSTRACT
Odd viscoelastic materials are constrained by fewer symmetries than their even counterparts. The breaking of these symmetries allows these materials to exhibit different features, which have attracted considerable attention in recent years. Immersing a bead in such complex fluids allows for probing their physical properties, highlighting signatures of their oddity and exploring the consequences of these broken symmetries. We present the conditions under which the activity of an odd viscoelastic fluid can give rise to linear instabilities in the motion of the probe particle, and we unveil how the features of the probe particle dynamics depend on the oddity and activity of the viscoelastic medium in which it is immersed.
ABSTRACT
How morphogenetic movements are robustly coordinated in space and time is a fundamental open question in biology. We study this question using the wing of Drosophila melanogaster, an epithelial tissue that undergoes large-scale tissue flows during pupal stages. Previously, we showed that pupal wing morphogenesis involves both cellular behaviors that allow relaxation of mechanical tissue stress, as well as cellular behaviors that appear to be actively patterned (Etournay et al., 2015). Here, we show that these active cellular behaviors are not guided by the core planar cell polarity (PCP) pathway, a conserved signaling system that guides tissue development in many other contexts. We find no significant phenotype on the cellular dynamics underlying pupal morphogenesis in mutants of core PCP. Furthermore, using laser ablation experiments, coupled with a rheological model to describe the dynamics of the response to laser ablation, we conclude that while core PCP mutations affect the fast timescale response to laser ablation they do not significantly affect overall tissue mechanics. In conclusion, our work shows that cellular dynamics and tissue shape changes during Drosophila pupal wing morphogenesis do not require core PCP as an orientational guiding cue.
Subject(s)
Drosophila Proteins , Drosophila , Animals , Drosophila/metabolism , Drosophila melanogaster/physiology , Drosophila Proteins/genetics , Drosophila Proteins/metabolism , Pupa/genetics , Wings, Animal/physiology , Morphogenesis/genetics , Cell Polarity , MutationABSTRACT
We investigate how randomly oriented cell traction forces lead to fluidization in a vertex model of epithelial tissues. We find that the fluidization occurs at a critical value of the traction force magnitude F_{c}. We show that this transition exhibits critical behavior, similar to the yielding transition of sheared amorphous solids. However, we find that it belongs to a different universality class, even though it satisfies the same scaling relations between critical exponents established in the yielding transition of sheared amorphous solids. Our work provides a fluidization mechanism through active force generation that could be relevant in biological tissues.
Subject(s)
Traction , EpitheliumABSTRACT
Liquid-liquid phase separation yields spherical droplets that eventually coarsen to one large, stable droplet governed by the principle of minimal free energy. In chemically fueled phase separation, the formation of phase-separating molecules is coupled to a fuel-driven, non-equilibrium reaction cycle. It thus yields dissipative structures sustained by a continuous fuel conversion. Such dissipative structures are ubiquitous in biology but are poorly understood as they are governed by non-equilibrium thermodynamics. Here, we bridge the gap between passive, close-to-equilibrium, and active, dissipative structures with chemically fueled phase separation. We observe that spherical, active droplets can undergo a morphological transition into a liquid, spherical shell. We demonstrate that the mechanism is related to gradients of short-lived droplet material. We characterize how far out of equilibrium the spherical shell state is and the chemical power necessary to sustain it. Our work suggests alternative avenues for assembling complex stable morphologies, which might already be exploited to form membraneless organelles by cells.
ABSTRACT
When a body moves through a fluid, it can experience a force orthogonal to its movement called lift force. Odd viscous fluids break parity and time-reversal symmetry, suggesting the existence of an odd lift force on tracer particles, even at vanishing Reynolds numbers and for symmetric geometries. It was previously found that an incompressible odd fluid cannot induce lift force on a tracer particle with no-slip boundary conditions, making signatures of odd viscosity in the two-dimensional bulk elusive. By computing the response matrix for a tracer particle, we show that an odd compressible fluid can produce an odd lift force. Using shell localization, we provide analytic expressions for the drag and odd lift forces acting on the tracer particle in a steady state and also at finite frequency. Importantly, we find that the existence of an odd lift force in a steady state requires taking into account the nonconservation of the fluid mass density due to the coupling between the two-dimensional surface and the three-dimensional bulk fluid.
ABSTRACT
Animal organs exhibit complex topologies involving cavities and tubular networks, which underlie their form and function1-3. However, how topology emerges during the development of organ shape, or morphogenesis, remains elusive. Here we combine tissue reconstitution and quantitative microscopy to show that tissue topology and shape is governed by two distinct modes of topological transitions4,5. One mode involves the fusion of two separate epithelia and the other involves the fusion of two ends of the same epithelium. The morphological space is captured by a single control parameter that can be traced back to the relative rates of the two epithelial fusion modes. Finally, we identify a pharmacologically accessible pathway that regulates the frequency of two modes of epithelial fusion, and demonstrate the control of organoid topology and shape. The physical principles uncovered here provide fundamental insights into the self-organization of complex tissues6.
ABSTRACT
The control of cell shape during cytokinesis requires a precise regulation of mechanical properties of the cell cortex. Only few studies have addressed the mechanisms underlying the robust production of unequal-sized daughters during asymmetric cell division. Here we report that unequal daughter-cell sizes resulting from asymmetric sensory organ precursor divisions in Drosophila are controlled by the relative amount of cortical branched Actin between the two cell poles. We demonstrate this by mistargeting the machinery for branched Actin dynamics using nanobodies and optogenetics. We can thereby engineer the cell shape with temporal precision and thus the daughter-cell size at different stages of cytokinesis. Most strikingly, inverting cortical Actin asymmetry causes an inversion of daughter-cell sizes. Our findings uncover the physical mechanism by which the sensory organ precursor mother cell controls relative daughter-cell size: polarized cortical Actin modulates the cortical bending rigidity to set the cell surface curvature, stabilize the division and ultimately lead to unequal daughter-cell size.
Subject(s)
Actins , Nuclear Family , Cytokinesis , Neurons , Stem CellsABSTRACT
Chemically active systems such as living cells are maintained out of thermal equilibrium due to chemical events which generate heat and lead to active fluctuations. A key question is to understand on which time and length scales active fluctuations dominate thermal fluctuations. Here, we formulate a stochastic field theory with Poisson white noise to describe the heat fluctuations which are generated by stochastic chemical events and lead to active temperature fluctuations. We find that on large length- and timescales, active fluctuations always dominate thermal fluctuations. However, at intermediate length- and timescales, multiple crossovers exist which highlight the different characteristics of active and thermal fluctuations. Our work provides a framework to characterize fluctuations in active systems and reveals that local equilibrium holds at certain length- and timescales.
ABSTRACT
Stress-strain constitutive relations in solids with an internal angular degree of freedom can be modeled using Cosserat (also called micropolar) elasticity. In this paper, we explore Cosserat materials that include chiral active components and hence odd elasticity. We calculate static elastic properties and show that the static response to rotational stresses leads to strains that depend on both Cosserat and odd elasticity. We compute the dispersion relations in odd Cosserat materials in the overdamped regime and find the presence of exceptional points. These exceptional points create a sharp boundary between a Cosserat-dominated regime of complete wave attenuation and an odd-elasticity-dominated regime of propagating waves. We conclude by showing the effect of Cosserat and odd-elasticity terms on the polarization of Rayleigh surface waves.
ABSTRACT
We show that dislocations in active two-dimensional (2D) smectic liquid crystals with underlying rotational symmetry are always unbound in the presence of noise, meaning the active smectic phase does not exist for nonzero noise in d=2. The active smectic phase can, like equilibrium smectics in 2D, be stabilized by applying rotational symmetry-breaking fields; however, even in the presence of such fields, active smectics are still much less stable against noise than equilibrium ones, when the symmetry-breaking field(s) are weak.
ABSTRACT
The kinetics of chemical reactions are determined by the law of mass action, which has been successfully applied to homogeneous, dilute mixtures. At nondilute conditions, interactions among the components can give rise to coexisting phases, which can significantly alter the kinetics of chemical reactions. Here, we derive a theory for chemical reactions in coexisting phases at phase equilibrium. We show that phase equilibrium couples the rates of chemical reactions of components with their diffusive exchanges between the phases. Strikingly, the chemical relaxation kinetics can be represented as a flow along the phase equilibrium line in the phase diagram. A key finding of our theory is that differences in reaction rates between coexisting phases stem solely from phase-dependent reaction rate coefficients. Our theory is key to interpreting how concentration levels of reactive components in condensed phases control chemical reaction rates in synthetic and biological systems.
Subject(s)
Kinetics , DiffusionABSTRACT
A key event at the onset of development is the activation of a contractile actomyosin cortex during the oocyte-to-embryo transition1-3. Here we report on the discovery that, in Caenorhabditis elegans oocytes, actomyosin cortex activation is supported by the emergence of thousands of short-lived protein condensates rich in F-actin, N-WASP and the ARP2/3 complex4-8 that form an active micro-emulsion. A phase portrait analysis of the dynamics of individual cortical condensates reveals that condensates initially grow and then transition to disassembly before dissolving completely. We find that, in contrast to condensate growth through diffusion9, the growth dynamics of cortical condensates are chemically driven. Notably, the associated chemical reactions obey mass action kinetics that govern both composition and size. We suggest that the resultant condensate dynamic instability10 suppresses coarsening of the active micro-emulsion11, ensures reaction kinetics that are independent of condensate size and prevents runaway F-actin nucleation during the formation of the first cortical actin meshwork.
Subject(s)
Actomyosin , Biomolecular Condensates , Caenorhabditis elegans , Oocytes , Actin Cytoskeleton/metabolism , Actin-Related Protein 2/metabolism , Actin-Related Protein 3/metabolism , Actins/metabolism , Actomyosin/chemistry , Actomyosin/metabolism , Animals , Biomolecular Condensates/chemistry , Biomolecular Condensates/metabolism , Caenorhabditis elegans/embryology , Caenorhabditis elegans/metabolism , Caenorhabditis elegans Proteins/metabolism , Emulsions/chemistry , Emulsions/metabolism , Oocytes/metabolism , Wiskott-Aldrich Syndrome Protein, Neuronal/metabolismABSTRACT
Morphogen gradients are a central concept in developmental biology. Their formation often involves the secretion of morphogens from a local source, that spread by diffusion in the cell field, where molecules eventually get degraded. This implies limits to both the time and length scales over which morphogen gradients can form which are set by diffusion coefficients and degradation rates. Towards the goal of identifying plausible mechanisms capable of extending the gradient range, we here use theory to explore properties of a cell-to-cell signaling relay. Inspired by the millimeter-scalewnt-expression and signaling gradients in flatworms, we consider morphogen-mediated morphogen production in the cell field. We show that such a relay can generate stable morphogen and signaling gradients that are oriented by a local, morphogen-independent source of morphogen at a boundary. This gradient formation can be related to an effective diffusion and an effective degradation that result from morphogen production due to signaling relay. If the secretion of morphogen produced in response to the relay is polarized, it further gives rise to an effective drift. We find that signaling relay can generate long-range gradients in relevant times without relying on extreme choices of diffusion coefficients or degradation rates, thus exceeding the limits set by physiological diffusion coefficients and degradation rates. A signaling relay is hence an attractive principle to conceptualize long-range gradient formation by slowly diffusing morphogens that are relevant for patterning in adult contexts such as regeneration and tissue turn-over.
Subject(s)
Models, Biological , Signal Transduction , Cell Communication , Diffusion , Morphogenesis/physiology , Signal Transduction/physiologyABSTRACT
Active chiral viscoelastic materials exhibit elastic responses perpendicular to the applied stresses, referred to as odd elasticity. We use a covariant formulation of viscoelasticity combined with an entropy production analysis to show that odd elasticity is not only present in active systems but also in broad classes of passive chiral viscoelastic fluids. In addition, we demonstrate that linear viscoelastic chiral solids require activity in order to manifest odd elastic responses. To model the phenomenon of passive odd viscoelasticity we propose a chiral extension of Jeffreys model. We apply our covariant formalism in order to derive the dispersion relations of hydrodynamic modes and obtain clear imprints of odd viscoelastic behavior.
ABSTRACT
SignificanceBiomolecular condensates are intracellular organelles that are not bounded by membranes and often show liquid-like, dynamic material properties. They typically contain various types of proteins and nucleic acids. How the interaction of proteins and nucleic acids finally results in dynamic condensates is not fully understood. Here we use optical tweezers and fluorescence microscopy to study how the prototypical prion-like protein Fused-in-Sarcoma (FUS) condenses with individual molecules of single- and double-stranded DNA. We find that FUS adsorbs on DNA in a monolayer and hence generates an effectively sticky FUS-DNA polymer that collapses and finally forms a dynamic, reversible FUS-DNA co-condensate. We speculate that protein monolayer-based protein-nucleic acid co-condensation is a general mechanism for forming intracellular membraneless organelles.