A Biomechanical Analysis of Ventral Furrow Formation in the Drosophila Melanogaster Embryo

The article provides a biomechanical analysis of ventral furrow formation in the Drosophila melanogaster embryo. Ventral furrow formation is the first large-scale morphogenetic movement in the fly embryo. It involves deformation of a uniform cellular monolayer formed following cellularisation, and has therefore long been used as a simple system in which to explore the role of mechanics in force generation. Here we use a quantitative framework to carry out a systematic perturbation analysis to determine the role of each of the active forces observed. The analysis confirms that ventral furrow invagination arises from a combination of apical constriction and apical–basal shortening forces in the mesoderm, together with a combination of ectodermal forces. We show that the mesodermal forces are crucial for invagination: the loss of apical constriction leads to a loss of the furrow, while the mesodermal radial shortening forces are the primary cause of the internalisation of the future mesoderm as the furrow rises. Ectodermal forces play a minor but significant role in furrow formation: without ectodermal forces the furrow is slower to form, does not close properly and has an aberrant morphology. Nevertheless, despite changes in the active mesodermal and ectodermal forces lead to changes in the timing and extent of furrow, invagination is eventually achieved in most cases, implying that the system is robust to perturbation and therefore over-determined

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. The technique involves image processing, segmenting of cells, grouping of cell outlines, calculation of dihedral planes, averaging along three-dimensional TJs, and matrix equation assembly and solution. The equations tend to be strongly overdetermined, allowing indistinct TJs to be ignored and solution error estimates to be determined. Application to clean and noisy synthetic data generated using Surface Evolver gave tension errors of 1.6?7%, and analyses of eight-cell murine embryos gave estimated errors smaller than the 10% uncertainty of companion aspiration experiments. Other possible areas of application include morphogenesis, cancer metastasis and tissue engineering.authorsversionPeer reviewe

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. Here, we have analyzed the role of differential TST for germ layer progenitor cell segregation during zebrafish gastrulation. Contrary to previous observations that differential TST drives germ layer progenitor cell segregation in vitro, we show that germ layers display indistinguishable TST within the gastrulating embryo, arguing against differential TST driving germ layer progenitor cell segregation in vivo. We further show that the osmolarity of the interstitial fluid (IF) is an important factor that influences germ layer TST in vivo, and that lower osmolarity of the IF compared with standard cell culture medium can explain why germ layers display differential TST in culture but not in vivo. Finally, we show that directed migration of mesendoderm progenitors is required for germ layer progenitor cell segregation and germ layer formation

Digital tracking algorithm reveals the influence of structural irregularities on joint movements in the human cervical spine

The final publication is available at Elsevier via https://dx.doi.org/10.1016/j.clinbiomech.2018.04.015 © 2018. This manuscript version is made available under the CC-BY-NC-ND 4.0 license https://creativecommons.org/licenses/by-nc-nd/4.0/Background Disc height loss and osteophytes change the local mechanical environment in the spine; while previous research has examined kinematic dysfunction under degenerative change, none has looked at the influence of disc height loss and osteophytes throughout movement. Methods Twenty patients with pain related to the head, neck or shoulders were imaged via videofluoroscopy as they underwent sagittal-plane flexion and extension. A clinician graded disc height loss and osteophytes as “severe/moderate”, “mild”, or “none”. A novel tracking algorithm quantified motions of each vertebra. This information was used to calculate intervertebral angular and shear displacements. The digital algorithm made it practical to track individual vertebrae in multiple patients through hundreds of images without bias. Findings Cases without height loss/osteophytes had a consistent increase in intervertebral angular displacement from C2/C3 to C5/C6, like that of healthy individuals, and mild height losses did not produce aberrations that were systematic or necessarily discernable. However, joints with moderate to severe disc height loss and osteophytes exhibited reduced range of motion compared to adjacent unaffected joints in that patient and corresponding joints in patients without structural irregularities. Interpretation Digitally-obtained motion histories of individual joints allowed anatomical joint changes to be linked with changes in joint movement patterns. Specifically, disc height loss and osteophytes were found to influence cervical spine movement in the sagittal plane, reducing angular motions at affected joints by approximately 10% between those with and without height loss and osteophytes. Further, these joint changes were associated with perturbed intervertebral angular and shear movements.Natural Sciences and Engineering Research Council (NSERC) Discovery Grant

A biomechanical analysis of ventral furrow formation in the Drosophila melanogaster embryo

The article provides a biomechanical analysis of ventral furrow formation in the Drosophila melanogaster embryo. Ventral furrow formation is the first large-scale morphogenetic movement in the fly embryo. It involves deformation of a uniform cellular monolayer formed following cellularisation, and has therefore long been used as a simple system in which to explore the role of mechanics in force generation. Here we use a quantitative framework to carry out a systematic perturbation analysis to determine the role of each of the active forces observed. The analysis confirms that ventral furrow invagination arises from a combination of apical constriction and apical–basal shortening forces in the mesoderm, together with a combination of ectodermal forces. We show that the mesodermal forces are crucial for invagination: the loss of apical constriction leads to a loss of the furrow, while the mesodermal radial shortening forces are the primary cause of the internalisation of the future mesoderm as the furrow rises. Ectodermal forces play a minor but significant role in furrow formation: without ectodermal forces the furrow is slower to form, does not close properly and has an aberrant morphology. Nevertheless, despite changes in the active mesodermal and ectodermal forces lead to changes in the timing and extent of furrow, invagination is eventually achieved in most cases, implying that the system is robust to perturbation and therefore over-determined

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. At early stages of the process, leading actin protrusions generate traction forces that point away from the wound, showing that wound closure is initially driven by cell crawling. At later stages, we observed unanticipated patterns of traction forces pointing towards the wound. Such patterns have strong force components that are both radial and tangential to the wound. We show that these force components arise from tensions transmitted by a heterogeneous actomyosin ring to the underlying substrate through focal adhesions. The structural and mechanical organization reported here provides cells with a mechanism to close the wound by cooperatively compressing the underlying substrate

The Mechanics of Metastasis: Insights from a Computational Model

<div><p>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. The present work focuses on the dissemination steps, from dissociation to circulation. The model shows that certain surface tension relationships must be satisfied for cancerous cells to dissociate from a primary tumour and that these equations are analogous to those that govern dissociation of embryonic cells. For a dissociated cell to then migrate by invadopodium extension and contraction and exhibit the shapes seen in experiments, the invadopodium must generate a contraction equal to approximately twice that produced by the interfacial tension associated with surrounding cells. Intravasation through the wall of a vessel is governed by relationships akin to those in the previous two steps, while release from the vessel wall is governed by equations that involve surface and interfacial tensions. The model raises a number of potential research questions. It also identifies how specific mechanical properties and the sub-cellular structural components that give rise to them might be changed so as to thwart particular metastatic steps and thereby block the spread of cancer.</p> </div

A simulation of the complete dissemination phase.

<p>All of the steps in the dissemination process are demonstrated in this simulation. In this simulation as in the others reported here unless noted otherwise, γ_NN  =  γ_TT  =  γ_WW  = 10, γ_NT  = 20, γ_NW  =  γ_WB  = 50, γ_CT  = 40, γ_CN  =  γ_CW  = 20, q = 2. The starting configuration is shown in (A). The metastasis journey begins when a single cell in the tumour becomes sufficiently discriminated from its neighbours that <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044281#pone.0044281.e002" target="_blank">Equation (2</a>) is satisfied (see text). It is then pushed out of the tumour by interfacial tension differences (B) until at (C), its contact with the tumour becomes vanishingly small. Changes in cell signalling associated with loss of contact with the primary tumour or contact only with stromal cells or detection of chemotactic gradients are then assumed to initiate invadopodia that are oriented toward a nearby blood vessel and that pull the cell toward it (D). The migrating cell encounters the outside surface of the blood vessel (E) and continues to advance by invadopodium action until it makes contact with the blood stream (F). A further programming change in the cell occurs (γ_CN increases to 60) and it is pushed into the blood stream (G) and released there (γ_CW increases to 80) (H) by surface and interfacial tension differences akin to those that pushed the cell out of the primary tumour. See also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044281#pone.0044281.s001" target="_blank">Movie S1</a>.</p

Construction of the finite element model.

<p>(A) Biochemical networks regulate the construction of signaling and structural proteins and lead to assembly and regulation of mechanical components. These mechanical components (especially actomyosin and adhesion systems) generate net interfacial tensions γ along the cell edges <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044281#pone.0044281-Lecuit1" target="_blank">[41]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044281#pone.0044281-Chen2" target="_blank">[42]</a> (B). Cancer cells (type “C”) generate invadopodia which push their way between the normal cells (labelled as type “N”). They then contract with a force assumed to be q times the tension γ_NN that acts along the boundaries of the surrounding cells. In the finite element model, the edge forces are generated using rods that lie along each edge and that have zero stiffness but carry a constant tension γ that is specific to the cell types that form the boundary. The viscosity µ of the contents of the cells is modeled using a series of orthogonal dashpots <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044281#pone.0044281-Brodland8" target="_blank">[46]</a>, only one set of which is illustrated in the figure.</p