Elongated Cells Drive Morphogenesis in a Surface-Wrapped Finite-Element Model of Germband Retraction.

During Drosophila embryogenesis, the germband first extends to curl around the posterior end of the embryo and then retracts back; however, retraction is not simply the reversal of extension. At a tissue level, extension is coincident with ventral furrow formation, and at a cellular level, extension occurs via convergent cell neighbor exchanges in the germband, whereas retraction involves only changes in cell shape. To understand how cell shapes, tissue organization, and cellular forces drive germband retraction, we investigate this process using a whole-embryo, surface-wrapped cellular finite-element model.

Coordination of Receptor Tyrosine Kinase Signaling and Interfacial Tension Dynamics Drives Radial Intercalation and Tube Elongation.

We sought to understand how cells collectively elongate epithelial tubes. We first used 3D culture and biosensor imaging to demonstrate that epithelial cells enrich Ras activity, phosphatidylinositol (3,4,5)-trisphosphate (PIP), and F-actin to their leading edges during migration within tissues. PIP enrichment coincided with, and could enrich despite inhibition of, F-actin dynamics, revealing a conserved migratory logic compared with single cells.

Interstitial fluid osmolarity modulates the action of differential tissue surface tension in progenitor cell segregation during gastrulation.

The segregation of different cell types into distinct tissues is a fundamental process in metazoan development. Differences in cell adhesion and cortex tension are commonly thought to drive cell sorting by regulating tissue surface tension (TST). However, the role that differential TST plays in cell segregation within the developing embryo is as yet unclear.

Inferring cellular forces from image stacks.

Although the importance of cellular forces to a wide range of embryogenesis and disease processes is widely recognized, measuring these forces is challenging, especially in three dimensions. Here, we introduce CellFIT-3D, a force inference technique that allows tension maps for three-dimensional cellular systems to be estimated from image stacks. Like its predecessors, video force microscopy and CellFIT, this cell mechanics technique assumes boundary-specific interfacial tensions to be the primary drivers, and it constructs force-balance equations based on triple junction (TJ) dihedral angles.

A videofluoroscopy-based tracking algorithm for quantifying the time course of human intervertebral displacements.

The motions of individual intervertebral joints can affect spine motion, injury risk, deterioration, pain, treatment strategies, and clinical outcomes. Since standard kinematic methods do not provide precise time-course details about individual vertebrae and intervertebral motions, information that could be useful for scientific advancement and clinical assessment, we developed an iterative template matching algorithm to obtain this data from videofluoroscopy images. To assess the bias of our approach, vertebrae in an intact porcine spine were tracked and compared to the motions of high-contrast markers.

How computational models can help unlock biological systems.

With computation models playing an ever increasing role in the advancement of science, it is important that researchers understand what it means to model something; recognize the implications of the conceptual, mathematical and algorithmic steps of model construction; and comprehend what models can and cannot do. Here, we use examples to show that models can serve a wide variety of roles, including hypothesis testing, generating new insights, deepening understanding, suggesting and interpreting experiments, tracing chains of causation, doing sensitivity analyses, integrating knowledge, and inspiring new approaches. We show that models can bring together information of different kinds and do so across a range of length scales, as they do in multi-scale, multi-faceted embryogenesis models, some of which connect gene expression, the cytoskeleton, cell properties, tissue mechanics, morphogenetic movements and phenotypes.

Non-straight cell edges are important to invasion and engulfment as demonstrated by cell mechanics model.

Computational models of cell-cell mechanical interactions typically simulate sorting and certain other motions well, but as demands on these models continue to grow, discrepancies between the cell shapes, contact angles and behaviours they predict and those that occur in real cells have come under increased scrutiny. To investigate whether these discrepancies are a direct result of the straight cell-cell edges generally assumed in these models, we developed a finite element model that approximates cell boundaries using polylines with an arbitrary number of segments. We then compared the predictions of otherwise identical polyline and monoline (straight-edge) models in a variety of scenarios, including annealing, single- and multi-cell engulfment, sorting, and two forms of mixing--invasion and checkerboard pattern formation.

Practical aspects of the cellular force inference toolkit (CellFIT).

If we are to fully understand the reasons that cells and tissues move and acquire their distinctive geometries during processes such as embryogenesis and wound healing, we will need detailed maps of the forces involved. One of the best current prospects for obtaining this information is noninvasive force-from-images techniques such as CellFIT, the Cellular Force Inference Toolkit, whose various steps are discussed here. Like other current quasistatic approaches, this one assumes that cell shapes are produced by interactions between interfacial tensions and intracellular pressures.

Forces driving epithelial wound healing.

A fundamental feature of multicellular organisms is their ability to self-repair wounds through the movement of epithelial cells into the damaged area. This collective cellular movement is commonly attributed to a combination of cell crawling and "purse-string" contraction of a supracellular actomyosin ring. Here we show by direct experimental measurement that these two mechanisms are insufficient to explain force patterns observed during wound closure.

CellFIT: a cellular force-inference toolkit using curvilinear cell boundaries.

Mechanical forces play a key role in a wide range of biological processes, from embryogenesis to cancer metastasis, and there is considerable interest in the intuitive question, "Can cellular forces be inferred from cell shapes?" Although several groups have posited affirmative answers to this stimulating question, nagging issues remained regarding equation structure, solution uniqueness and noise sensitivity. Here we show that the mechanical and mathematical factors behind these issues can be resolved by using curved cell edges rather than straight ones. We present a new package of force-inference equations and assessment tools and denote this new package CellFIT, the Cellular Force Inference Toolkit.

On the origins of the mitotic shift in proliferating cell layers.

Background: During plant and animal development, monolayer cell sheets display a stereotyped distribution of polygonal cell shapes. In interphase cells these shapes range from quadrilaterals to decagons, with a robust average of six sides per cell. In contrast, the subset of cells in mitosis exhibits a distinct distribution with an average of seven sides.

Modeling cell elongation during germ band retraction: cell autonomy versus applied anisotropic stress.

The morphogenetic process of germ band retraction in embryos involves coordinated movements of two epithelial tissues - germ band and amnioserosa. The germ band shortens along its rostral-caudal or head-to-tail axis, widens along its perpendicular dorsal-ventral axis, and uncurls from an initial 'U' shape. The amnioserosa mechanically assists this process by pulling on the crook of the U-shaped germ band.

The mechanics of metastasis: insights from a computational model.

Although it may seem obvious that mechanical forces are required to drive metastatic cell movements, understanding of the mechanical aspects of metastasis has lagged far behind genetic and biochemical knowledge. The goal of this study is to learn about the mechanics of metastasis using a cell-based finite element model that proved useful for advancing knowledge about the forces that drive embryonic cell and tissue movements. Metastasis, the predominant cause of cancer-related deaths, involves a series of mechanical events in which one or more cells dissociate from a primary tumour, migrate through normal tissue, traverse in and out of a multi-layer circulatory system vessel and resettle.

Measuring the modulus of silicone hydrogel contact lenses.

Purpose: The purpose of this study is to demonstrate a novel method for measuring the modulus of contact lenses in their as-received, variable-thickness form and to determine whether modulus varies with location within commercial lenses and whether it is dependent on lens geometry and temperature.

Methods: The thickness profiles of lenses having powers from -8 diopters (D) to +4 D were measured using a Rehder electronic thickness gauge. Strip-shaped specimens having a width of 5.

Assessing the mechanical energy costs of various tissue reshaping mechanisms.

Early-stage embryos must reshape the tissues of which they are made into organs and other life-sustaining structures; and if non-mammalian embryos fail to complete these tasks before the energy provided by their yolk runs out, they die. The aim of this study is to use a cell-level computational model to investigate the energetic cost of a variety of mechanisms that can drive an in-plane reshaping pattern known as convergent extension--a motif in which a tissue narrows in one in-plane direction and expands in another. Mechanisms considered include oriented lamellipodia, directed mitosis, stress fibers, and anisotropic external tension.

DRhoGEF2 regulates cellular tension and cell pulsations in the Amnioserosa during Drosophila dorsal closure.

Coordination of apical constriction in epithelial sheets is a fundamental process during embryogenesis. Here, we show that DRhoGEF2 is a key regulator of apical pulsation and constriction of amnioserosal cells during Drosophila dorsal closure. Amnioserosal cells mutant for DRhoGEF2 exhibit a consistent decrease in amnioserosa pulsations whereas overexpression of DRhoGEF2 in this tissue leads to an increase in the contraction time of pulsations.

A framework for connecting gene expression to morphogenetic movements in embryos.

Although much has been learned about genetic networks, cell mechanics, and whole-embryo mechanics through experimental and computational studies, the challenge of connecting these separate bodies of knowledge into an integrated whole remains. Here, we offer a multiscale biochemical-mechanical framework from which such integration might proceed. We identify components of the framework for which quantitative descriptions are currently available, and use the framework to gain insight into convergent extension and gastrulation--crucial tissue movements that occur in early stage amphibian embryos.

Control of the mitotic cleavage plane by local epithelial topology.

For nearly 150 years, it has been recognized that cell shape strongly influences the orientation of the mitotic cleavage plane (e.g., Hofmeister, 1863).

Video force microscopy reveals the mechanics of ventral furrow invagination in Drosophila.

The absence of tools for mapping the forces that drive morphogenetic movements in embryos has impeded our understanding of animal development. Here we describe a unique approach, video force microscopy (VFM), that allows detailed, dynamic force maps to be produced from time-lapse images. The forces at work in an embryo are considered to be decomposed into active and passive elements, where active forces originate from contributions (e.

From genes to neural tube defects (NTDs): insights from multiscale computational modeling.

The morphogenetic movements, and the embryonic phenotypes they ultimately produce, are the consequence of a series of events that involve signaling pathways, cytoskeletal components, and cell- and tissue-level mechanical interactions. In order to better understand how these events work together in the context of amphibian neurulation, an existing multiscale computational model was augmented. Geometric data for this finite element-based mechanical model were obtained from 3D surface reconstructions of live axolotl embryos and serial sections of fixed specimens.

Identifying same-cell contours in image stacks: a key step in making 3D reconstructions.

Identification of contours belonging to the same cell is a crucial step in the analysis of confocal stacks and other image sets in which cell outlines are visible, and it is central to the making of 3D cell reconstructions. When the cells are close packed, the contour grouping problem is more complex than that found in medical imaging, for example, because there are multiple regions of interest, the regions are not separable from each other by an identifiable background and regions cannot be distinguished by intensity differences. Here, we present an algorithm that uses three primary metrics-overlap of contour areas in adjacent images, co-linearity of the centroids of these areas across three images in a stack, and cell taper-to assign cells to groups.

Cinemechanometry (CMM): A method to determine the forces that drive morphogenetic movements from time-lapse images.

Although cell-level mechanical forces are crucial to tissue self-organization in contexts ranging from embryo development to cancer metastases to regenerative engineering, the absence of methods to map them over time has been a major obstacle to new understanding. Here, we present a technique for constructing detailed, dynamic maps of the forces driving morphogenetic events from time-lapse images. Forces in the cell are considered to be separable into unknown active driving forces and known passive forces, where actomyosin systems and microtubules contribute primarily to the first group and intermediate filaments and cytoplasm to the latter.

Cellular interfacial and surface tensions determined from aggregate compression tests using a finite element model.

Although previous studies suggested that the interfacial tension gamma(cc) acting along cell-cell boundaries and the effective viscosity mu of the cell cytoplasm could be measured by compressing a spherical aggregate of cells between parallel plates, the mechanical understanding necessary to extract this information from these tests-tests that have provided the surface tension sigma(cm) acting along cell-medium interfaces-has been lacking. These tensions can produce net forces at the subcellular level and give rise to cell motions and tissue reorganization, the rates of which are regulated by mu. Here, a three-dimensional (3D) cell-based finite element model provides insight into the mechanics of the compression test, where these same forces are at work, and leads to quantitative relationships from which the effective viscosity mu of the cell cytoplasm, the tension gamma(cc) that acts along internal cell-cell interfaces and the surface tension sigma(cp) along the cell-platen boundaries can be determined from force-time curves and aggregate profiles.

Combining laser microsurgery and finite element modeling to assess cell-level epithelial mechanics.

Laser microsurgery and finite element modeling are used to determine the cell-level mechanics of the amnioserosa-a morphogenetically crucial epithelium on the dorsal surface of fruit fly embryos (Drosophila melanogaster). In the experiments, a tightly focused laser ablates a subcellular hole (1 microm in diameter) that passes clean through the epithelium. The surrounding cells recoil from the wound site with a large range of initial recoil velocities.

Detecting mitoses in time-lapse images of embryonic epithelia using intensity analysis.

Although the frequency and orientation of mitoses can significantly affect the mechanics of early embryo development, these data have not been available due to a shortage of suitable automated techniques. Fluorescence imaging, though popular, requires biochemical intervention and is not always possible or desirable. Here, a new technique that takes advantage of a localized intensity change that occurs in bright field images is used to identify mitoses.