Computer simulation of cellular movements: cell-sorting, cellular migration through a mass of cells and contact inhibition

The motility rules for cellular movement proposed earlier by Goel &; Rogers for engulfment of two or more intact embryonic tissues have been used to simulate on a computer the phenomena of cell-sorting, migration of individual cells through a mass of cells and contact inhibition of overlapping. These simulations in the most part are found to be consistent with the observations with real cells.     

Moving forward moving backward: directional sorting of chemotactic cells due to size and adhesion differences

Differential movement of individual cells within tissues is an important yet poorly understood process in biological development. Here we present a computational study of cell sorting caused by a combination of cell adhesion and chemotaxis, where we assume that all cells respond equally to the chemotactic signal. To capture in our model mesoscopic properties of biological cells, such as their size and deformability, we use the Cellular Potts Model, a multiscale, cell-based Monte Carlo model. We demonstrate a rich array of cell-sorting phenomena, which depend on a combination of mescoscopic cell properties and tissue level constraints. Under the conditions studied, cell sorting is a fast process, which scales linearly with tissue size. We demonstrate the occurrence of “absolute negative mobility”, which means that cells may move in the direction opposite to the applied force (here chemotaxis). Moreover, during the sorting, cells may even reverse the direction of motion. Another interesting phenomenon is “minority sorting”, where the direction of movement does not depend on cell type, but on the frequency of the cell type in the tissue. A special case is the cAMP-wave-driven chemotaxis of Dictyostelium cells, which generates pressure waves that guide the sorting. The mechanisms we describe can easily be overlooked in studies of differential cell movement, hence certain experimental observations may be misinterpreted.     

Contact inhibition of what? An analytical review

Quite a number of phenomena having to do with cells' influences upon one another's movements have come to be regarded as expressions of “contact inhibition.” However, no single, central mechanism has been shown to underlie them all. Consequently, the term “contact inhibition” should not be used without operational modifiers. Inhibitions of individual cell movements imputed to be mediated by cell-cell contacts include inhibition of overlapping (which results in monolayering), of colony expansion, of cell speed (nuclear translocation), of ruffling, of orthogonal movement (proposed to explain spontaneous parallel alignment of cells), and of neighbor exchanges. The six inhibitions listed above are operationally distinct, and only two (overlapping and colony expansion) are known to result from a common mechanism. A seventh phenomenon, so-called “contact inhibition of cell division” (more operationally termed postconfluence inhibition of cell division) is in a separate category and is not considered here. Evidence eliminating action-at-a-distance is available only for the first three, and hence only these should at present be termed contact inhibitions. Inhibition of neighbor exchanges is yet hypothetical; at its extreme, it would immobilize cells in a confluent monolayer, but such immobilization has been found not to occur. Contact inhibition of overlapping, the most studied of the six, is not displayed by invasive cells with respect to normal cells; invasive tumor cells overlap freely upon normal cells, although not necessarily upon one another. Contact inhibition of overlapping, and its loss by invasive cells, can readily be interpreted, by means of the differential adhesion hypothesis, as consequences of cell-type-specific differences in cell-cell and cell-substratum “strengths of adhesion.” These strengths of adhesion are formulated as specific interfacial free energies, which are the only parameters of cellular adhesiveness that have been rigorously shown to determine equilibrium configurations of cell populations.     

The mechanics of heterotypic cell aggregates: insights from computer simulations

Finite element-based computer simulations are used to investigate a number of phenomena, including tissue engulfment, cell sorting, and checkerboard-pattern formation, exhibited by heterotypic cell aggregates. The simulations show that these phenomena can be driven by a single equivalent force, namely a surface (or interfacial) tension, that results from cytoskeletal components and cell-cell adhesions. They also reveal that tissue engulfment, cell sorting, and checkerboard-pattern formation involve several discernible mechanical features or stages. With the aid of analytical arguments, we identify the conditions necessary for each of these phenomena. These findings are consistent with previous experimental investigations and computer simulations, but pose significant challenges to current theories of cell sorting and tissue engulfment.     

Vav regulates activation of Rac but not Cdc42 during FcgammaR-mediated phagocytosis

Phagocytosis is the process whereby cells direct the spatially localized, receptor-driven engulfment of particulate materials. It proceeds via remodeling of the actin cytoskeleton and shares many of the core cytoskeletal components involved in adhesion and migration. Small GTPases of the Rho family have been widely implicated in coordinating actin dynamics in response to extracellular signals and during diverse cellular processes, including phagocytosis, yet the mechanisms controlling their recruitment and activation are not known. We show herein that in response to ligation of Fc receptors for IgG (FcgammaR), the guanine nucleotide exchange factor Vav translocates to nascent phagosomes and catalyzes GTP loading on Rac, but not Cdc42. The Vav-induced Rac activation proceeds independently of Cdc42 function, suggesting distinct roles for each GTPase during engulfment. Moreover, inhibition of Vav exchange activity or of Cdc42 activity does not prevent Rac recruitment to sites of particle attachment. We conclude that Rac is recruited to Fcgamma membrane receptors in its inactive, GDP-bound state and that Vav regulates phagocytosis through subsequent catalysis of GDP/GTP exchange on Rac.     

On a possible relationship between bacterial endospore formation and the origin of eukaryotic cells

The acquisition of intracellular organelles, including mitochondria and plastids and a membrane-bounded nucleus, have been postulated to be key events in the development of the eukaryotic from the prokaryotic ancestral cell. The two major hypotheses to account for such acquisitions are: (1) primitive cells originally obtained organelles by engulfing free-living prokaryotes which then entered into symbiotic association (“endosymbiosis”) with them; (2) organelles arose through the engulfment by the primitive cell of part of its own cytoplasm. To some extent, the former hypothesis has received most support, because endosymbiosis is known to occur in extant organisms, whilst the latter hypothesis has received less support, because cytoplasmic engulfment by prokaryotes is not now thought to occur. However, during the process of endospore formation by extant bacteria, the protoplast within the single cell is observed to divide in a unique manner such that the cell in effect engulfs a portion of its own cytoplasm. The process is strikingly similar to the engulfment suggested by the second hypothesis to have initiated the evolution of eukaryotes. The engulfed cytoplasm is bounded by a double membrane within the “mother cell” and contains enzymes, ribosomes and a complete genome. In many respects this parallels the supposed primitive eukaryotic state and, it is argued, confers potential advantages on the cell, particularly through the control that the “mother cell” can exert on the enclosed compartment. It is hypothesized that bacterial endospore formation is therefore one product of evolution from an early engulfment event that led also to the development of complex eukaryotic cells.     

A guide and guard: The many faces of T-cadherin

Cadherins are a superfamily of transmembrane proteins that mediate calcium-dependent intercellular adhesion. T-cadherin (T-cad, H-cadherin or cadherin-13) is an atypical member, lacking transmembrane and cytosolic domains and possessing a glycosylphosphatidylinositol moiety that anchors T-cadherin to the plasma membrane. This article reviews current knowledge on the biomolecular characteristics of T-cadherin, its expression and function in different tissues in health and disease and its mechanisms of signal transduction. The structural characteristics of T-cadherin protein predict that it is unlikely to function as a “true” adhesion molecule in vivo. Studies from different fields suggest that it may act rather as a signalling receptor participating in recognition of the environment and regulation of cell motility, proliferation and phenotype. Cellular expression levels of T-cadherin in various tissues frequently correlate (be it negatively or positively) with the proliferative potential of the cells. Loss- and gain-of-function studies demonstrate the ability of T-cadherin to modulate cell motility and growth. Gathering evidence suggests that the “functional predestination” of T-cadherin is in control of tissue architecture through “guiding” navigation of moving structures, segregating functional tissue compartments and “guarding” integrity of functionally connected tissue layers.     

Differentiation,ageing, and terminal differentiation: A semantic analysis

The largely unsolved problems in the theoretical analysis of differentiation and ageing involve a substantial component of linguistic (semantic) difficulties. Some of these are simple traps of ambiguity, resulting from metaphorical or analogical employment of established terms—for example, “terminal differentiation” (loss of division potential in vitro) as a borrowing from “differentiation” as used by developmental biologists, or “commitment” by analogy with “determination”. Some difficulties represent a failure to adopt (at least provisionally) an operational (empirical) view—for example, failure to ask what is the nature of the evidence for the view that a fertilized ovum is totipotential, or to scrutinize the evidence for the view that cells “terminally differentiated” in vitro in a conventional medium are in fact moribund under all conditions, or to examine more closely the view that the differentiated state and the cycling state are mutually exclusive.With respect to the problem of ageing, we review some of the critical experiments on ”terminal differentiation” or “clonal senescence”. We then proceed to consider some of the models that have been proposed, including a molecular model proposed by the author which appears to overcome some of the objections to other models. Some of the models exemplify the results of what are ultimately semantic vices.The problems with which these remarks began should indeed yield to the immense and novel resources of molecular biology. But the development of complete analyses demands not only good luck and delicate technique but also critical semantic clarity and severity. Given the best tools, we shall solve major theoretical problems only if we understand quite fully what problem it is that we are trying to solve—and the history of science illustrates that this is not as elementary a matter as it sounds. For the working scientist semantics (and indeed all philosophy of science) is not an indulgence or a frill, but a basic technical resource.     

Les marques de croissance osseuse comme indicateurs de l‘âge chez les lézards

Abstract From a comparative viewpoint, this paper deals with structural and functional aspects of skeletal annulations in lizards, especially those displayed by sections cut transversally through the larger bones of the appendages. Cross-sections of the diaphyses exhibit an alternation of concentrically arranged thinner and thicker annulations which were formed during colder and warmer seasons, respectively. The thinner annulations are chromophile and anisotropic but not hypermineralized; they are generally referred to as “resting lines” or “adhesion lines” and are due to a slowing down of the temperature-dependent growth process; they also differ structurally from “cementing reversal lines”. In certain cases, the thinner annulations (here called LAC) are surrounded by bone lamellae and thus compare closely with the annuli on the flat bones. From the study of two lizards of known age, as well as on the basis of a preliminary experimentation with fluorochrom labelling, it has been demonstrated conclusively that the recurrence of the LAC in lacertilians shows an annual regularity. Therefore, it would seem that the LAC can be employed for age determination in lizards.     

The morphology of the central nervous system of the barnacle,Semibalanus cariosus (Pallas)

The central nervous system of the sessile barnacle, Semibalanus cariosus (Pallas), has been studied with the particular aim of determining the locations of neuron somata in relation to peripheral nerves. This was accomplished by tracing peripheral nerves using dissection and methylene blue staining techniques, histological methods, and by permitting cobaltous chloride to diffuse via axons into ganglia (“backfilling”). The neuron maps resulting from the study reveal some well-defined sub-systems, a considerable degree of functional clumping of neuron somata, and some unexpected projections of neurons in the CNS. Neurophysiological studies based on these findings are in progress.     

On the Phenomenon of “Perezhivanie”

The author presents a short historical review of the concept of “perezhivanie” beginning with L.S. Vygotsky. She compares this concept with reflection as two central concepts revealing a person's consciousness. A systemic picture of these two phenomena is elaborated by introducing a hierarchy of reflection. A further step is made that relates the “perezhivanie—reflection” system to (psychoanalytic) self-formations. Empirical studies shed light on phenomena connected to “perezhivanie.”     

A model for the genetic control of differential adhesion in insect epidermis

Adhesive relations among cells are believed to play a major role in determining patterns of serial homology, of intercalary regeneration, and of neuronal connectivity. Models for the genetic control of adhesion during development can provide a framework for further analysis of these phenomena. Investigators studying development of Drosophila have proposed that differentiation of segments and of imaginal discs is controlled by a set of bistable “selector genes”. In each region the settings of the selector genes form a binary “word” which determines the properties of cells in the region, including their adhesiveness. I have made an explicit proposal for the relation between binary words and adhesiveness, by assuming that active selector genes repress synthesis of “adhesor” macromolecules, which promote adhesion. This hypothesis correctly predicts the relative cohesiveness of cells in four pupal tissues of the moth Manduca. Works of cohesion and adhesion among the four cell types are deduced from published results of grafting experiments by modelling insect epidermis as a viscoelastic fluid.Further comparisons between deductions from the genetic and fluid models suggest that selector genes, or the adhesor molecules they regulate, interact within single cells in determining adhesiveness between cells. From a specific version of the genetic model I deduce that pairwise interactions between selector genes or adhesor molecules can determine many, though not all, of the relative works of adhesion between unlike cells in Manduca. The genetic and fluid models thus provide a set of working hypotheses for predicting patterns of intercellular adhesion in insect epidermis and for analyzing results of experiments designed to test such predictions.     

The relation of root systems to shoot systems in vascular plants

The roots and shoots of vascular plants may be positionally and developmentally related in various ways. However, botanical teaching and research are strongly influenced by the paradigmatic annual dicotyledon, whose bipolar embryo develops into a plant with root and shoot meeting only at the hypocotyl. In 1930 Goebel criticized this example as a general model for plants, proposing instead the opposed concepts “allorhizy” (referring to plants whose root and shoot are related as above) and “homorhizy”(referring to plants without a bipolar embryo, all of whose roots are shoot-borne, e.g., pteridophytes). Goebel’s approach permeates the extensive German morphological literature, but has been virtually ignored in English-language literature. The allorhizy/homorhizy dichotomy has proved heuristic. However, it suggests a correlation between embryo type and mature morphology that does not always hold. Furthermore, it does not take into account the root-borne shoots typical of many plant species. Finally, Goebel’s presentation of the terms (which he does not explicitly define) creates ambiguity as to whether they designate structural concepts or the attributes of evolutionary groups. The alternative proposed here is a structural analysis of the possible topological relationships among root and shoot systems. Each structural class is then considered with regard to embryo types, potential for clonal growth and other ecological correlates, and phylogenetic distribution. This approach provides both a test of Goebel’s concepts and a basis for further comparative study of wholeplant form.     

Different mechanisms drive the maintenance of polymorphism at loci subject to strong versus weak fluctuating selection

The long‐running debate about the role of selection in maintaining genetic variation has been given new impetus by the discovery of hundreds of seasonally oscillating polymorphisms in wild Drosophila, possibly stabilized by an alternating summer‐winter selection regime. Historically, there has been skepticism about the potential of temporal variation to balance polymorphism, because selection must be strong to have a meaningful stabilizing effect—unless dominance also varies over time (“reversal of dominance”). Here, we develop a simplified model of seasonally variable selection that simultaneously incorporates four different stabilizing mechanisms, including two genetic mechanisms (“cumulative overdominance” and reversal of dominance), as well as ecological “storage” (“protection from selection” and boom‐bust demography). We use our model to compare the stabilizing effects of these mechanisms. Although reversal of dominance has by far the greatest stabilizing effect, we argue that the three other mechanisms could also stabilize polymorphism under plausible conditions, particularly when all three are present. With many loci subject to diminishing returns epistasis, reversal of dominance stabilizes many alleles of small effect. This makes the combination of the other three mechanisms, which are incapable of stabilizing small effect alleles, a better candidate for stabilizing the detectable frequency oscillations of large effect alleles.     

Apoptotic cells are cleared by directional migration and elmo1- dependent macrophage engulfment

Apoptotic cell death is essential for development and tissue homeostasis. Failure to clear apoptotic cells can ultimately cause inflammation and autoimmunity. Apoptosis has primarily been studied by staining of fixed tissue sections, and a clear understanding of the behavior of apoptotic cells in living tissue has been elusive. Here, we use a newly developed technique to track apoptotic cells in real time as they emerge and are cleared from the zebrafish brain. We find that apoptotic cells are remarkably motile, frequently migrating several cell diameters to the periphery of living tissues. F-actin remodeling occurs in surrounding cells, but also within the apoptotic cells themselves, suggesting a cell-autonomous component of motility. During the first 2 days of development, engulfment is rare, and most apoptotic cells lyse at the brain periphery. By 3 days postfertilization, most cell corpses are rapidly engulfed by macrophages. This engulfment requires the guanine nucleotide exchange factor elmo1. In elmo1-deficient macrophages, engulfment is rare and may occur through macropinocytosis rather than directed engulfment. These findings suggest that clearance of apoptotic cells in living vertebrates is accomplished by the combined actions of apoptotic cell migration and elmo1-dependent macrophage engulfment.     

IrreC/rst-mediated cell sorting during Drosophila pupal eye development depends on proper localisation of DE-cadherin

Remodelling of tissues depends on the coordinated regulation of multiple cellular processes, such as cell-cell communication, differential cell adhesion and programmed cell death. During pupal development, interommatidial cells (IOCs) of the Drosophila eye initially form two or three cell rows between individual ommatidia, but then rearrange into a single row of cells. The surplus cells are eliminated by programmed cell death, and the definitive hexagonal array of cells is formed, which is the basis for the regular pattern of ommatidia visible in the adult eye. Here, we show that this cell-sorting process depends on the presence of a continuous belt of the homophilic cell adhesion protein DE-cadherin at the apical end of the IOCs. Elimination of this adhesion belt by mutations in shotgun, which encodes DE-cadherin, or its disruption by overexpression of DE-cadherin, the intracellular domain of Crumbs, or by a dominant version of the monomeric GTPase Rho1 prevents localisation of the transmembrane protein IrreC-rst to the border between primary pigment cells and IOCs. As a consequence, the IOCs are not properly sorted and supernumerary cells survive. During the sorting process, Notch-mediated signalling in IOCs acts downstream of DE-cadherin to restrict IrreC-rst to this border. The data are discussed in relation to the roles of selective cell adhesion and cell signalling during tissue reorganisation.     

Schistosoma mansoni: mechanism of cercarial tail loss and its significance to host penetration

The mechanism of tail loss from cercariae of Schistosoma mansoni was investigated under experimental conditions. Tail loss from 90% or more of cercariae occurred in less than 10 min when packed organisms were incubated at 30 C in a minimal volume of water. Proteolytic secretions from the acetabular glands did not play a significant part in this process since the addition of known protease inhibitors nor the addition of secretions collected from other cercariae influenced the rate of the process. Tail loss was inhibited when the packed cercariae were incubated in saline at concentrations of 0.05 M or above though glandular secretion occurred at an equal rate in both water and saline. Tail loss was also inhibited by the chelating agents ethylenediaminotetraacetic acid (EDTA) and ethyleneglycoltetraacetic acid (EGTA). A marked feature of cercariae packed in water was their clumping and adhesion to the walls of the containing vessel by secretions from the postacetabular glands. Such clumping did not occur in saline or chelant solutions. It is suggested that the most probable mechanism for tail loss is a simple mechanical trauma effected by the movement of the tail acting against the resistance of the secretion-fixed body.During incubation under conditions which remove the “coat” from the body of the cercaria, the “coat” of the tail remains intact.     

A statistical mechanical model for hydrogen exchange in globular proteins.

We develop a statistical mechanical theory for the mechanism of hydrogen exchange in globular proteins. Using the HP lattice model, we explore how the solvent accessibilities of chain monomers vary as proteins fluctuate from their stable native conformations. The model explains why hydrogen exchange appears to involve two mechanisms under different conditions of protein stability; (1) a “global unfolding” mechanism by which all protons exchange at a similar rate, approaching that of the denatured protein, and (2) a “stable-state” mechanism by which protons exchange at rates that can differ by many orders of magnitude. There has been some controversy about the stable-state mechanism: does exchange take place inside the protein by solvent penetration, or outside the protein by the local unfolding of a subregion? The present model indicates that the stable-state mechanism of exchange occurs through an ensemble of conformations, some of which may bear very little resemblance to the native structure. Although most fluctuations are small-amplitude motions involving solvent penetration or local unfolding, other fluctuations (the conformational distant relatives) can involve much larger transient excursions to completely different chain folds.     

Correlation “waves” in brain-like structures

A correspondence is established between a tangible model of brain structure (and function) and a system of observer-observed interactions. The observed quantities are “stimuli” in the form of signal amplitude distributions in a mass of neuron-like units; the observer is a set of neurons (not circumscribed in a local region) in which a distributed parameter mirrors the stimulus history of the set, i.e., represents a “memory”. Utilizing the theory of the Perceptron, a contemporary brain model, it is demonstrated that large systems composed of many observer-observed interactions exhibit quantum mechanical behavior on a “macroscopic” scale. This behavior entails wave-like phenomena and the need of applying the superposition mechanics to system information content calculations.