Biomedical subjects

Evan A Zamir

Publications and source records attributed to Evan A Zamir.

7 recordsLinked to original sources

PubMed2006

Mesodermal cell displacements during avian gastrulation are due to both individual cell-autonomous and convective tissue movements.

Gastrulation is a fundamental process in early development that results in the formation of three primary germ layers. During avian gastrulation, presumptive mesodermal cells in the dorsal epiblast ingress through a furrow called the primitive streak (PS), and subsequently move away from the PS and form adult tissues. The biophysical mechanisms driving mesodermal cell movements during gastrulation in amniotes, notably warm-blooded embryos, are not understood. Until now, a major challenge has been distinguishing local individual cell-autonomous (active) displacements from convective displacements caused by large-scale (bulk) morphogenetic tissue movements. To address this problem, we used multiscale, time-lapse microscopy and a particle image velocimetry method for computing tissue displacement fields. Immunolabeled fibronectin was used as an in situ marker for quantifying tissue displacements. By imaging fluorescently labeled mesodermal cells and surrounding extracellular matrix simultaneously, we were able to separate directly the active and passive components of cell displacement during gastrulation. Our results reveal the following: (i) Convective tissue motion contributes significantly to total cell displacement and must be subtracted to measure true cell-autonomous displacement; (ii) Cell-autonomous displacement decreases gradually after regression from the PS; and (iii) There is an increasing cranial-to-caudal (head-to-tail) cell-autonomous motility gradient, with caudal cells actively moving away from the PS faster than cranial cells. These studies show that, in some regions of the embryo, total mesodermal cell displacements are mostly due to convective tissue movements; thus, the data have profound implications for understanding cell guidance mechanisms and tissue morphogenesis in warm-blooded embryos.

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PubMed2006

Extracellular matrix macroassembly dynamics in early vertebrate embryos.

This chapter focuses on the in vivo macroassembly dynamics of fibronectin and fibrillin-2--two prominent extracellular matrix (ECM) components, present in vertebrate embryos at the earliest stages of development. The ECM is an inherently dynamic structure with a well-defined position fate: ECM filaments are not only anchored to and move with established tissue boundaries, but are repositioned prior to the formation of new anatomical features. We distinguish two ECM filament relocation processes-each operating on different length scales. First, ECM filaments are moved by large-scale tissue motion, which rearranges major organ primordia within the embryo. The second type of motion, on the scale of the individual ECM filaments, is driven by local motility and protrusive activity of nearby cells. The motion decomposition is made practically possible by recent advances in microscopy and high-resolution particle image velocimetry algorithms. We demonstrate that both kinds of motion contribute substantially to the establishment of normal ECM structure, and both must be taken into account when attempting to understand ECM macroassembly during embryonic morphogenesis. The tissue-scale motion changes the local amount (density) and the tissue-level structure (e.g., orientation) of ECM fibers. Local reorganization includes filament assembly and the segregation of ECM into specific patterns. Local reorganization takes place most actively at Hensen's node and around the primitive streak. These regions are also sites of active cell migration, where fibrillin-2 and fibronectin are often colocalized in ECM globules, and new fibrillin-2 foci are deposited. During filament assembly, the globular patches of ECM are joined into larger linear structures in a hierarchical process: increasingly larger structures are created by the aggregation of smaller units. A future understanding of ECM assembly thus requires the study of the complex interactions between biochemical assembly steps, local cell action, and tissue motion.

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PubMed2005

A digital image-based method for computational tissue fate mapping during early avian morphogenesis.

The early stages of vertebrate development, encompassing gastrulation, segmentation, and caudal axis formation, presumably involve large (finite) morphogenetic deformations; however, there are few quantitative biomechanical data available for describing such large-scale or tissue-level deformations in the embryo. In this study, we present a new method for automated computational "tissue fate mapping," by combining a recently developed high-resolution time-lapse digital microscopy system for whole-avian embryo imaging with particle image velocimetry (PIV), a well-established digital image correlation technique for measuring continuum deformations. Tissue fate mapping, as opposed to classical cell fate mapping or other cell tracking methods, is used to track the spatiotemporal trajectories of arbitrary (virtual) tissue material points in various layers of the embryo, which can then be used to calculate finite morphogenetic deformation or strain maps. To illustrate the method, we present representative tissue fate and strain mapping data for normal early-stage quail embryos. These data demonstrate, to our knowledge, for the first time, large tissue-level deformations that are shared between different germ layers in the embryo, suggesting a more global morphogenetic patterning mechanism than had been previously appreciated.

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PubMed2004

Dynamic imaging of cell, extracellular matrix, and tissue movements during avian vertebral axis patterning.

Vertebrate axis patterning depends on cell and extracellular matrix (ECM) repositioning and proper cell-ECM interactions. However, there are few in vivo data addressing how large-scale tissue deformations are coordinated with the motion of local cell ensembles or the displacement of ECM constituents. Combining the methods of dynamic imaging and experimental biology allows both cell and ECM fate-mapping to be correlated with ongoing tissue deformations. These fate-mapping studies suggest that the axial ECM components "move" both as a composite meshwork and as autonomous particles, depending on the length scale being examined. Cells are also part of this composite, and subject to passive displacements resulting from tissue deformations. However, in contrast to the ECM, cells are self-propelled. The net result of cell and ECM displacements, along with proper ECM-cell adhesion, is the assembly of new tissue architecture. Data herein show that disruption of normal cell-ECM interactions during axis formation results in developmental abnormalities and a disorganization of the ECM. Our goal in characterizing the global displacement patterns of axial cells and ECM is to provide critical information regarding existing strain fields in the segmental plate and paraxial mesoderm. Deducing the mechanical influences on cell behavior is critical, if we are to understand vertebral axis patterning. Supplementary material for this article is available online at http://www.mrw.interscience.wiley.com/suppmat/1542-975X/suppmat/72/v72.266.html.

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PubMed2004

On the effects of residual stress in microindentation tests of soft tissue structures.

Microindentation methods are commonly used to determine material properties of soft tissues at the cell or even sub-cellular level. In determining properties from force-displacement (FD) data, it is often assumed that the tissue is initially a stress-free, homogeneous, linear elastic half-space. Residual stress, however, can strongly influence such results. In this paper, we present a new microindentation method for determining both elastic properties and residual stress in soft tissues that, to a first approximation, can be regarded as a pre-stressed layer embedded in or adhered to an underlying relatively soft, elastic foundation. The effects of residual stress are shown using two linear elastic models that approximate specific biological structures. The first model is an axially loaded beam on a relatively soft, elastic foundation (i.e., stress-fiber embedded in cytoplasm), while the second is a radially loaded plate on a foundation (e.g., cell membrane or epithelium). To illustrate our method, we use a nonlinear finite element (FE) model and experimental FD and surface contour data to find elastic properties and residual stress in the early embryonic chick heart, which, in the region near the indenter tip, is approximated as an isotropic circular plate under tension on a foundation. It is shown that the deformation of the surface in a microindentation test can be used along with FD data to estimate material properties, as well as residual stress, in soft tissue structures that can be regarded as a plate under tension on an elastic foundation. This method may not be as useful, however, for structures that behave as a beam on a foundation.

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PubMed2004

Material properties and residual stress in the stage 12 chick heart during cardiac looping.

During the morphogenetic process of cardiac looping, the initially straight cardiac tube bends and twists into a curved tube. The biophysical mechanisms that drive looping remain unknown, but the process clearly involves mechanical forces. Hence, it is important to determine mechanical properties of the early heart, which is a muscle-wrapped tube consisting primarily of a thin outer layer of myocardium surrounding a thick extracellular matrix compartment known as cardiac jelly. In this work, we used microindentation experiments and finite element modeling, combined with an inverse computational method, to determine constitutive relations for the myocardium and cardiac jelly at the outer curvature of stage 12 chick hearts. Material coefficients for exponential strain-energy density functions were found by fitting force-displacement and surface displacement data near the indenter Residual stress in the myocardium also was estimated. These results should be useful for computational models of the looping heart.

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PubMed2003

Mechanical asymmetry in the embryonic chick heart during looping.

Cardiac looping, which begins with ventral bending and rightward rotation of the primitive heart tube, is an essential morphogenetic event that occurs early in vertebrate development. The biophysical mechanism that drives this process is unknown. It has been speculated that increased stiffness along the dorsal side of the ventricle combined with an intrinsic cardiac force causes the heart to bend. There is no experimental support for this hypothesis, however, since little is known about regional mechanical properties of the heart during looping. We directly measured diastolic stiffness of the inner curvature (IC), outer curvature (OC), and dorsal-ventral sides of the stage 12 chick heart by microindentation. The IC of intact hearts was found to be significantly stiffer than either the OC or the sides. which were of similar stiffness. Isolated cardiac jelly, which is a thick, extracellular matrix compartment underlying the myocardium, was approximately an order of magnitude softer than intact hearts. The results of a computational model simulating the indentation experiments, combined with the stiffness measurements, suggests the regional variation in stiffness is due to the material properties of the myocardium. A second model shows that a relatively stiff IC may facilitate bending of the heart tube during looping.

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