ABSTRACT
Formation of fluid-filled lumina by epithelial tissues is essential for organ development. How cells control the hydraulic and cortical forces to control lumen morphology is not well understood. Here, we quantified the mechanical role of tight junctions in lumen formation using MDCK-II cysts. We found that the paracellular ion barrier formed by claudin receptors is not required for the hydraulic inflation of a lumen. However, the depletion of the zonula occludens scaffold resulted in lumen collapse and folding of apical membranes. Combining quantitative measurements of hydrostatic lumen pressure and junctional tension with modeling enabled us to explain lumen morphologies from the pressure-tension force balance. Tight junctions promote lumen inflation by decreasing cortical tension via the inhibition of myosin. In addition, our results suggest that excess apical area contributes to lumen opening. Overall, we provide a mechanical understanding of how epithelial cells use tight junctions to modulate tissue and lumen shape.
ABSTRACT
Spatiotemporal coordination of chromatin and subnuclear compartments is crucial for cells. Numerous enzymes act inside nucleus-some of those transiently link and pass two chromatin segments. Here, we study how such an active perturbation affects fluctuating dynamics of an inclusion in the chromatic medium. Using numerical simulations and a versatile effective model, we categorize inclusion dynamics into three distinct modes. The transient-link-and-pass activity speeds up inclusion dynamics by affecting a slow mode related to chromatin remodeling, viz., size and shape of the chromatin meshes.
Subject(s)
Chromatin Assembly and Disassembly , Chromatin , Cell NucleusABSTRACT
Cell proliferation is a fundamental process underlying embryogenesis, homeostasis, wound healing, and cancer. The process involves multiple events during each cell cycle, such as cell growth, contractile ring formation, and division to daughter cells, which affect the surrounding cell population geometrically and mechanically. However, existing methods do not comprehensively describe the dynamics of multicellular structures involving cell proliferation at a subcellular resolution. In this study, we present a novel model for proliferative multicellular dynamics at the subcellular level by building upon the nonconservative fluid membrane (NCF) model that we developed in earlier research. The NCF model utilizes a dynamically-rearranging closed triangular mesh to depict the shape of each cell, enabling us to analyze cell dynamics over extended periods beyond each cell cycle, during which cell surface components undergo dynamic turnover. The proposed model represents the process of cell proliferation by incorporating cell volume growth and contractile ring formation through an energy function and topologically dividing each cell at the cleavage furrow formed by the ring. Numerical simulations demonstrated that the model recapitulated the process of cell proliferation at subcellular resolution, including cell volume growth, cleavage furrow formation, and division to daughter cells. Further analyses suggested that the orientation of actomyosin stress in the contractile ring plays a crucial role in the cleavage furrow formation, i.e., circumferential orientation can form a cleavage furrow but isotropic orientation cannot. Furthermore, the model replicated tissue-scale multicellular dynamics, where the successive proliferation of adhesive cells led to the formation of a cell sheet and stratification on the substrate. Overall, the proposed model provides a basis for analyzing proliferative multicellular dynamics at subcellular resolution.
Subject(s)
Actin Cytoskeleton , Cell Division , Cell MembraneABSTRACT
The forces that cells, tissues, and organisms exert on the surface of a soft substrate can be measured using Traction Force Microscopy (TFM), an important and well-established technique in Mechanobiology. The usual TFM technique (two-dimensional, 2D TFM) treats only the in-plane component of the traction forces and omits the out-of-plane forces at the substrate interfaces (2.5D) that turn out to be important in many biological processes such as tissue migration and tumour invasion. Here, we review the imaging, material, and analytical tools to perform "2.5D TFM" and explain how they are different from 2D TFM. Challenges in 2.5D TFM arise primarily from the need to work with a lower imaging resolution in the z-direction, track fiducial markers in three-dimensions, and reliably and efficiently reconstruct mechanical stress from substrate deformation fields. We also discuss how 2.5D TFM can be used to image, map, and understand the complete force vectors in various important biological events of various length-scales happening at two-dimensional interfaces, including focal adhesions forces, cell diapedesis across tissue monolayers, the formation of three-dimensional tissue structures, and the locomotion of large multicellular organisms. We close with future perspectives including the use of new materials, imaging and machine learning techniques to continuously improve the 2.5D TFM in terms of imaging resolution, speed, and faithfulness of the force reconstruction procedure.
Subject(s)
Mechanical Phenomena , Traction , Microscopy, Atomic Force/methods , Focal Adhesions , Stress, Mechanical , Cell AdhesionABSTRACT
Cell adhesion plays an important role in a wide range of biological situations, including embryonic development, cancer invasion, and wound healing. Although several computational models describing adhesion dynamics have been proposed, models applicable to long-term, large-length-scale cell dynamics are lacking. In this study we investigated possible states of long-term adherent cell dynamics in three-dimensional space by constructing a continuum model of interfacial interactions between adhesive surfaces. In this model a pseudointerface is supposed between each pair of triangular elements that discretize cell surfaces. By introducing a distance between each pair of elements, the physical properties of the interface are given by interfacial energy and friction. The proposed model was implemented into the model of a nonconservative fluid cell membrane where the cell membrane dynamically flows with turnover. Using the implemented model, numerical simulations of adherent cell dynamics on a substrate under flow were performed. The simulations not only reproduced the previously reported dynamics of adherent cells, such as detachment, rolling, and fixation on the substrate, but also discovered other dynamic states, including cell slipping and membrane flow patterns, corresponding to behaviors that occur on much longer timescales than the dissociation of adhesion molecules. These results illustrate the variety of long-term adherent cell dynamics, which are more diverse than the short-term ones. The proposed model can be extended to arbitrarily shaped membranes, thus being useful for the mechanical analysis of a wide range of long-term cell dynamics where adhesion is essential.
Subject(s)
Friction , Cell Adhesion , Cell MembraneABSTRACT
On a curved surface, epithelial cells can adapt to geometric constraints by tilting and by exchanging their neighbors from apical to basal sides, known as an apico-basal topological transition 1 (AB-T1). The relationship between cell tilt, AB-T1s, and tissue curvature still lacks a unified understanding. Here, we propose a general framework for cell packing in curved environments and explain the formation of AB-T1s from the perspective of strain anisotropy. We find that steep curvature gradients can lead to cell tilting and induce AB-T1s. Alternatively, pressure differences across the epithelial tissue can drive AB-T1s in regions of large curvature anisotropy. The two mechanisms compete to determine the impact of tissue geometry and mechanics on optimized cell rearrangements in three dimensions.
Subject(s)
Epithelial Cells , EpitheliumABSTRACT
The number of cells in tissues is controlled by cell division and cell death, and its misregulation could lead to pathological conditions such as cancer. To maintain the cell numbers, a cell-elimination process called apoptosis also stimulates the proliferation of neighboring cells. This mechanism, apoptosis-induced compensatory proliferation, was originally described more than 40 years ago. Although only a limited number of the neighboring cells need to divide to compensate for the apoptotic cell loss, the mechanisms that select cells to divide have remained elusive. Here, we found that spatial inhomogeneity in Yes-associated protein (YAP)-mediated mechanotransduction in neighboring tissues determines the inhomogeneity of compensatory proliferation in Madin-Darby canine kidney (MDCK) cells. Such inhomogeneity arises from the non-uniform distribution of nuclear size and the non-uniform pattern of mechanical force applied to neighboring cells. Our findings from a mechanical perspective provide additional insight into how tissues precisely maintain homeostasis.
Subject(s)
Apoptosis , Mechanotransduction, Cellular , Animals , Dogs , Apoptosis/physiology , Cell Death , Cell Division , Madin Darby Canine Kidney Cells , Cell Proliferation/physiologyABSTRACT
Spatial organization of chromatin plays a critical role in genome regulation. Previously, various types of affinity mediators and enzymes have been attributed to regulate spatial organization of chromatin from a thermodynamics perspective. However, at the mechanistic level, enzymes act in their unique ways and perturb the chromatin. Here, we construct a polymer physics model following the mechanistic scheme of Topoisomerase-II, an enzyme resolving topological constraints of chromatin, and investigate how it affects interphase chromatin organization. Our computer simulations demonstrate Topoisomerase-II's ability to phase separate chromatin into eu- and heterochromatic regions with a characteristic wall-like organization of the euchromatic regions. We realized that the ability of the euchromatic regions to cross each other due to enzymatic activity of Topoisomerase-II induces this phase separation. This realization is based on the physical fact that partial absence of self-avoiding interaction can induce phase separation of a system into its self-avoiding and non-self-avoiding parts, which we reveal using a mean-field argument. Furthermore, motivated from recent experimental observations, we extend our model to a bidisperse setting and show that the characteristic features of the enzymatic activity-driven phase separation survive there. The existence of these robust characteristic features, even under the non-localized action of the enzyme, highlights the critical role of enzymatic activity in chromatin organization.
Subject(s)
Chromatin , Genome , Interphase , DNA Topoisomerases, Type II/genetics , PolymersABSTRACT
Controlling the patterns formed by self-propelled particles through dynamic self-organization is a challenging task. Although varieties of patterns associated with chiral self-propelled particles have been reported, essential factors that determine the morphology of the patterns have remained unclear. Here, we explore theoretically how torque formed upon collision of the particles affects the dynamic self-organization of the particles and determine the patterns. Based on a particle-based model with collision-induced torque and torque associated with self-propulsion, we find that introducing collision-induced torque turns the homogeneous bi-directionally aligned particles into rotating mono-polar flocks, which helps resolve a discrepancy in the earlier observations in microfilament gliding assays.
ABSTRACT
Tissue layers can generally slide at the interface, accompanied by the dissipation due to friction. Nevertheless, it remains elusive how force could propagate in a tissue with such interfacial friction. Here, we elaborate the force dynamics in a prototypical multilayer system in which an epithelial monolayer was cultivated upon an elastic substrate in contact with a hard surface, and discover a novel mechanism of pronounced force propagation over a long distance due to interfacial dynamics between substrate layers. We derived an analytical model for the dynamics of the elastic substrate under the shear stress provided by the cell layer at the surface boundary and the friction at bottom. The model reveals that sliding between substrate layers leads to an expanding stretch regime from a shear regime of substrate deformation in time and space. The regime boundary propagating diffusively with a speed depending on the stiffness, thickness, and slipperiness of the substrate, is a robust nature of a deformed elastic sheet with interfacial friction. These results shed new light on force propagation in tissues and our model could serve as a basis for studies of such propagation in a more complex tissue environment.
Subject(s)
Mechanical Phenomena , Friction , Elasticity , Stress, MechanicalABSTRACT
Living cells actively deform and move by their force generations in three-dimensional (3D) space. These 3D cell dynamics occur over a long-term time scale, ranging from tens of minutes to days. On such a time scale, turnover of cell membrane constituents due to endocytosis and exocytosis cannot be ignored, i.e., the surface membrane dynamically deforms without mass conservation. Although membrane turnover is essential for large deformation of cells, there is no computational framework yet to simulate long-term cell dynamics with a non-conservative fluidic membrane. In this paper, we proposed a computational framework for simulating the long-term dynamics of a cell membrane in 3D space. For this purpose, in the proposed framework, the cell surface membrane is treated as a viscous fluid membrane without mass conservation. Cell shape is discretized by a triangular mesh, and its dynamics are expressed by effective energy and dissipation function. The mesh structure, distorted by membrane motion, is dynamically optimized by introducing a modified dynamic remeshing method. To validate the proposed framework, numerical simulations were performed, showing that the membrane flow is reproduced in a physically consistent manner and that the artificial effects of the remeshing method were negligible. To further demonstrate the applicability of the proposed framework, numerical simulations of cell migration induced by a mechanism similar to the Marangoni effect, i.e., the polarized surface tension actively generated by the cell, were performed. The observed cell behaviors agreed with existing analytical solutions, indicating that the proposed computational framework can quantitatively reproduce long-term active cell dynamics with membrane turnover. Based on the simple description of cell membrane dynamics, this framework provides a useful basis for analyzing various cell shaping and movement.
Subject(s)
Metabolic Networks and Pathways , Cell Membrane/metabolism , Cell Shape , Computer Simulation , ViscosityABSTRACT
Lumens, liquid-filled cavities surrounded by polarized tissue cells, are elementary units involved in the morphogenesis of organs. Theoretical modeling and computations, which can integrate various factors involved in biophysics of morphogenesis of cell assembly and lumens, may play significant roles to elucidate the mechanisms in formation of such complex tissue with lumens. However, up to present, it has not been documented well what computational approaches or frameworks can be applied for this purpose and how we can choose the appropriate approach for each problem. In this review, we report some typical lumen morphologies and basic mechanisms for the development of lumens, focusing on three keywords - mechanics, hydraulics and geometry - while outlining pros and cons of the current main computational strategies. We also describe brief guidance of readouts, i.e., what we should measure in experiments to make the comparison with the model's assumptions and predictions.
Subject(s)
Cell Polarity , MorphogenesisABSTRACT
Understanding how migrating cells can establish both dynamic structures and coherent dynamics may provide mechanistic insights to study how living systems acquire complex structures and functions. Recent studies revealed that intercellular contact communication plays a crucial role for establishing cellular dynamic self-organization (DSO) and provided a theoretical model of DSO for migrating solitary cells in a free space. However, to apply those understanding to situations in living organisms, we need to know the role of cell-cell communication for tissue dynamics under spatial confinements and epithelial integrity. Here, we expand the previous numerical studies on DSO to migrating cells subjected spatial confinement and/or epithelial integrity. An epithelial monolayer is simulated by combining the model of cellular DSO and the cellular vertex model in two dimensions for apical integrity. Under confinement to a small space, theoretical models of both solitary and epithelial cells exhibit characteristic coherent dynamics, including apparent swirling. We also find that such coherent dynamics can allow the cells to overcome the strong constraint due to spatial confinement and epithelial integrity. Furthermore, we demonstrate how epithelial cell clusters behave without spatial confinement and find various cluster dynamics, including spinning, migration and elongation.
Subject(s)
Cell Communication , Models, Biological , Cell Movement , Epithelial CellsABSTRACT
Flocking is a fascinating coordinated behavior of living organisms or self-propelled particles (SPPs). Particularly, monopolar flocking has been attractive due to its potential applications in various fields. However, the underlying mechanism behind flocking and emergence of monopolar motion in flocking of SPPs has remained obscured. Here, we demonstrate monopolar flocking of kinesin-driven microtubules, a self-propelled biomolecular motor system. Microtubules with an intrinsic structural chirality preferentially move towards counter-clockwise direction. At high density, the CCW motion of microtubules facilitates monopolar flocking and formation of a spiral pattern. The monopolar flocking of microtubules is accounted for by a torque generated when the motion of microtubules was obstructed due to collisions. Our results shed light on flocking and emergence of monopolar motion in flocking of chiral active matters. This work will help regulate the polarity in collective motion of SPPs which in turn will widen their applications in nanotechnology, materials science and engineering.
Subject(s)
Kinesins/chemistry , Microtubules/chemistry , Nanotechnology , Animals , Motion , Particle Size , SwineABSTRACT
Symmetric or asymmetric positioning of intracellular structures including the nucleus and mitotic spindle steers various biological processes such as cell migration, division, and embryogenesis. In typical animal cells, both a sparse actomyosin meshwork in the cytoplasm and a dense actomyosin cortex underneath the cell membrane participate in the intracellular positioning. However, it remains unclear how these coexisting actomyosin structures regulate the positioning symmetry. To reveal the potential mechanism, we construct an in vitro model composed of cytoplasmic extracts and nucleus-like clusters confined in droplets. Here we find that periodic centripetal actomyosin waves contract from the droplet boundary push clusters to the center in large droplets, while network percolation of bulk actomyosin pulls clusters to the edge in small droplets. An active gel model quantitatively reproduces molecular perturbation experiments, which reveals that the tug-of-war between two distinct actomyosin networks with different maturation time-scales determines the positioning symmetry.
Subject(s)
Actomyosin/chemistry , Actomyosin/metabolism , Cell Division , Ovum/cytology , Actomyosin/genetics , Animals , Biomechanical Phenomena , Cell Membrane/chemistry , Cell Membrane/genetics , Cell Membrane/metabolism , Cell Size , Cytoplasm/chemistry , Cytoplasm/metabolism , Ovum/chemistry , XenopusABSTRACT
Biophysical mechanisms underlying collective cell migration of eukaryotic cells have been studied extensively in recent years. One mechanism that induces cells to correlate their motions is contact inhibition of locomotion, by which cells migrating away from the contact site. Here, we report that tail-following behavior at the contact site, termed contact following locomotion (CFL), can induce a non-trivial collective behavior in migrating cells. We show the emergence of a traveling band showing polar order in a mutant Dictyostelium cell that lacks chemotactic activity. We find that CFL is the cell-cell interaction underlying this phenomenon, enabling a theoretical description of how this traveling band forms. We further show that the polar order phase consists of subpopulations that exhibit characteristic transversal motions with respect to the direction of band propagation. These findings describe a novel mechanism of collective cell migration involving cell-cell interactions capable of inducing traveling band with polar order.
The cells of animals and many other living things are able to migrate together in groups. This collective cell migration plays crucial roles in many processes in animals such as forming organs and limbs, and healing wounds. A soil-dwelling amoeba called Dictyostelium discoideum or just Dicty for short is commonly used as a model to study how groups of cells migrate collectively. Individual Dicty cells may live alone but sometimes many cells come together to form a larger mobile structure called a "slug". Chemical signals coordinate how the cells collectively migrate to form the multicellular slug. Mutant Dicty cells that lack these chemical signal processes can still move together as a band that travels across a surface. This movement resembles a type of collective motion that has previously been observed in physics experiments using self-propelled particles. However, it remains unclear how this collective behavior works. Hayakawa et al. have now combined genetics, cell biology and computational approaches to study how groups of the mutant Dicty cells migrate together. The experiments showed that the traveling band is dynamically maintained by cells joining or leaving, and that this turnover is caused by simple interactions between the cells known as "contact following locomotion". Contact following locomotion has been also reported in mammalian cells so the findings of Hayakawa et al. may aid research into how animals develop and how errors in cell migration may lead to diseases. Further studies are required to find out whether other cells showing contact following locomotion also travel in a band.
Subject(s)
Cell Communication , Cell Movement , Contact Inhibition , Dictyostelium/physiology , Dictyostelium/genetics , Microscopy, Video , Models, Biological , Mutation , Single-Cell Analysis , Time FactorsABSTRACT
Emergence and collapse of coherent motions of self-propelled particles are affected more by particle motions and interactions than by their material or biological details. In the reconstructed systems of biofilaments and molecular motors, several types of collective motion including a global-order pattern emerge due to the alignment interaction. Meanwhile, earlier studies show that the alignment interaction of a binary collision of biofilaments is too weak to form the global order. The multiple collision is revealed to be important to achieve global order, but it is still unclear what kind of multifilament collision is actually involved. In this study, we demonstrate that not only alignment but also crossing of two filaments is essential to produce an effective multiple-particle interaction and the global order. We design the reconstructed system of biofilaments and molecular motors to vary a probability of the crossing of biofilaments on a collision and thus control the effect of volume exclusion. In this system, biofilaments glide along their polar strands on the turf of molecular motors and can align themselves nematically when they collide with each other. Our experiments show the counterintuitive result, in which the global order is achieved only when the crossing is allowed. When the crossing is prohibited, the cluster pattern emerges instead. We also investigate the numerical model in which we can change the strength of the volume exclusion effect and find that the global orientational order and clusters emerge with weak and strong volume exclusion effects, respectively. With those results and simple theory, we conclude that not only alignment but also finite crossing probability are necessary for the effective multiple-particles interaction forming the global order. Additionally, we describe the chiral symmetry breaking of a microtubule motion which causes a rotation of global alignment.
ABSTRACT
This Letter investigates what forms of cellular dynamic self-organization are caused through intercellular contact communication based on a theoretical model in which migrating cells perform contact following and contact inhibition and attraction of locomotion. Tuning those strengths causes varieties of dynamic patterns. This further includes a novel form of collective migration, snakelike dynamic assembly. Scrutinizing this pattern reveals that cells in this state can accurately respond to an external directional cue but have no spontaneous global polar order.
Subject(s)
Cell Communication/physiology , Cell Movement/physiology , Models, BiologicalABSTRACT
Exclusive interactions between self-driven objects may play crucial roles in their collective behavior, e.g., in collective migration of living cells. Here, such collective behavior is studied based on a simple but sufficient model taking account the exclusion effects, which incorporate the following two distinct kinds of exclusion interactions in two dimensions: The first is the mechanical exclusion wherein two objects mechanically repel each other when they overlap. The second is the scattering exclusion, wherein the directions along which each object tries to move are modulated to avoid overlapping. We propose a theoretical model based on two principles: (1) Each object maintains its own polarity with a fixed strength and attempts to move into the polarity direction and (2) objects interact with each other through the abovementioned exclusions. Based on this model, we look at the difference of consequences and combinatory effects of these two kinds of exclusions. Furthermore, we calculate the polar order of polarity directions without an external directional bias. Our results suggest that the combination of these two kinds of exclusions leads to effectively inelastic scattering of two objects, which eventually gives rise to global polar ordering. We also find that the traveling band can arise by this mechanism of alignment at the intermediate density, as generally seen in collective motion with polar alignment and investigated in various earlier works. Characteristics of transitions among disordered, traveling band, and homogeneously ordered states of the presented model are investigated, and their similarities and differences with those given by the explicit alignment interaction are discussed.
ABSTRACT
During animal development, epithelial cells forming a monolayer sheet move collectively to achieve the morphogenesis of epithelial tissues. One driving mechanism of such collective cell movement is junctional remodeling, which is found in the process of clockwise rotation of Drosophila male terminalia during metamorphosis. However, it still remains unknown how the motions of cells are spatiotemporally organized for collective movement by this mechanism. Since these moving cells undergo elastic deformations, the influence of junctional remodeling may mechanically propagate among them, leading to spatiotemporal pattern formations. Here, using a numerical cellular vertex model, we found that the junctional remodeling in collective cell movement exhibits spatiotemporal self-organization without requiring spatial patterns of molecular signaling activity. The junctional remodeling propagates as a wave in a specific direction with a much faster speed than that of cell movement. Such propagation occurs in both the absence and presence of fluctuations in the contraction of cell boundaries.