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1.
Flotillins control zebrafish epiboly through their role in cadherin‐mediated cell–cell adhesion
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Eduardo A. Rios Morris Stéphane Bodin Bénédicte Delaval Franck Comunale Virginie Georget Manoel L. Costa Georges Lutfalla Cécile Gauthier‐Rouvière 《Biology of the cell / under the auspices of the European Cell Biology Organization》2017,109(5):210-221
Zebrafish gastrulation and particularly epiboly that involves coordinated movements of several cell layers is a dynamic process for which regulators remain to be identified. We show here that Flotillin 1 and 2, ubiquitous and highly conserved proteins, are required for epiboly. Flotillins knockdown compromised embryo survival, strongly delayed epiboly and impaired deep cell radial intercalation and directed collective migration without affecting enveloping layer cell movement. At the molecular level, we identified that Flotillins are required for the formation of E‐cadherin‐mediated cell–cell junctions. These results provide the first in vivo evidence that Flotillins regulate E‐cadherin‐mediated cell–cell junctions to allow epiboly progression. 相似文献
2.
TETSUO KAGEYAMA 《Development, growth & differentiation》1985,27(5):633-638
The embryonic surface of the teleost, Oryzias latipes , was observed by scanning electron microscopy (SEM) to examine the last phase of epiboly or blastopore closure. The surface of the external yolk syncytial layer (E–YSL), a surface cytoplasmic layer encompassing the yolk sphere situated beyond the blastoderm, was highly undulated with surface folds of random orientation throughout most of epiboly (st. 14–20). Scattered microvilli were observed on the surface of the margin of the yolk plug in st. 18–20. The microvilli, 1 to 6 μm in length, were projected in a bunch at the end of blastopore closure (st. 20–21). The appearance of these microvilli in the last phase of epiboly is discussed with respect to the mechanism of epiboly. 相似文献
3.
We have examined the active collective movement of ectodermal cells from early gastrula of Xenopus laevis towards the point source of stretching, using techniques of videomicroscopy and scanning electron microscopy. We define this mode of cell movement as tensotaxis. This movement begins near the source of tension 5-10 min after the beginning of stretching and is spread in a relay fashion to more distant cells. As a result, a considerable fraction of observed cells more towards the source of stretching over a considerable territory at a rate of 0.6-3 mu/min. Subsequently, these movements are replaced by cell intercalation roughly oriented in the direction transverse to that of tissue stretching. It is proposed that tensotaxis is initiated by asymmetric deformation of the embryonic tissue due to the concentration (focusing) of a stretching force and contains both passive and active components. Data are presented supporting the view that, during normal development, tensotaxis may determine the movement of embryonic cells towards the blastopore and can also participate in other morphogenetic processes. 相似文献
4.
Influence of the relaxation of mechanical tensions upon collective cell movements, shape formation, and expression patterns
of tissue-specific genes has been studied in Xenopus laevis embryos. We show that the local relaxation of tensile stresses within the suprablastoporal area (SBA) performed at the early-midgastrula
stage leads to a complete arrest of normal convergent cell intercalation towards the dorsal midline. As a result, SBA either
remains nondeformed or protrudes a strip of cells migrating ventralwards along one of the lateral lips of the opened blastopore.
Already, few minutes later, the tissues in the ventral lip vicinity undergo abnormal transversal contraction/longitudinal
extension resulting in the abnormal cell convergence toward ventral (rather than dorsal) embryo midline. Within a day, the
dorsally relaxed embryos acquire pharyngula-like shapes and often possess tail-like protrusions. Their antero-posterior and
dorso-ventral polarity, as well as expression patterns of pan-neural (Sox3), muscular cardiac actin, and forebrain (Otx2)
genes substantially deviate from the normal ones. We suggest that normal gastrulation is permanently controlled by mechanical
stresses within the blastopore circumference. The role of tissue tensions in regulating collective cell movements and creating
pharyngula-like shapes are discussed. 相似文献
5.
D. O. Kiryukhin L. A. Shustikova E. E. Kopantseva N. N. Luchinskaya A. V. Belyavskii 《Biophysics》2010,55(6):921-925
The development of multicellular organisms is a complicated coordinated process of the movement of groups of embryonic cells,
which is controlled by many regulatory systems. At present little is known about the regulation of the earliest manifestations
of movement in embryogenesis: epiboly and radial intercalation. The coordinators of these processes may be small GTPases of
the Rho family and their activators, guanine nucleotide exchange factors. It has been shown in this work that overexpression
of a guanine nucleotide exchange factor xLARG in Xenopus laevis embryos leads to an increase in the amount of the active form of xLARG. In addition, an increase in the expression of xLARG
disturbs the process of radial intercalation. The data obtained suggest that xLARG is involved in maintaining the xLARG activation
level necessary for the occurrence of epiboly. 相似文献
6.
TETSUO KAGEYAMA 《Development, growth & differentiation》1980,22(4):659-668
A modification of silver nitrate staining, when applied to embryos of Oryzias latipes , was found to make clear not only the boundary of enveloping layer during epiboly, but also the outline of individual cells of the layer. The linear speed of advance of the enveloping layer was constant at fixed temperatures (20°, 25° or 30°C), except for the start and the end of the epibloy. Observation of the shape and arrangement of cells in the layer stained at successive stages of epiboly revealed that the enveloping layer expands uniformly over the yolk until late gastrula (3/4 epiboly). No cytokinetic figure was observed during epiboly until the blastopore was going to close. Total cell number of the layer remained constant during epiboly. Thus the expansion of the enveloping layer is accomplished without an accompanying increase in the number of constituent cells. In the last phase of epiboly, the surface area occupied by individual cells reduced locally at the region above the embryonic body, which suggests the occurrence in teleost of the convergence of cell sheets commonly observed in amphibian embryos. 相似文献
7.
Sandwich explants of the suprablastoporal area of Xenopus early-mid gastrula and same stages of entire embryos were stretched with two needles perpendicular to the direction of natural elongation of the axial rudiments. The changes in the embryonic shape and histological structure were monitored as well as the arrangement of descendants of one of dorsal blastomers labeled with fluorescein-dextran at the 16-cell stage. A substantial fraction of stretched explants reoriented along the applied stretch direction. The arrangement dynamics of fluorescein-dextran-labeled cells and explant shape demonstrate that this is an active response based on convergent intercalation of cells induced by stretching. Stretched gastrulae demonstrated arrested gastrulation, dorsoventral extension of the blastopore, and ventral flow of labeled cells towards the lateral lips of the blastopore, which was also mediated by convergent intercalation and tensotaxis. The obtained data are discussed in terms of the hypothesis of mechanical stress hyper-restoration. 相似文献
8.
There are two main approaches to unraveling the mechanisms involved in the regulation of collective cell movement. On the one hand, “in vitro” tests try to represent “in vivo” conditions. On the other hand, “in silico” tests aim to model this movement through the use of complex numerically implemented mathematical methods. This paper presents a simple cell-based mathematical model to represent the collective movement phenomena. This approach is used to better understand the different interactive forces which guide cell movement, focusing mainly on the role of the cell propulsion force with the substrate. Different applications are simulated for 2D cell cultures, wound healing, and collective cell movement in substrates with different degrees of stiffness. The model provides a plausible explanation of how cells work together in order to regulate their movement, showing the significant influence of the propulsive force exerted by the cell to the substrate on guiding the collective cell movement and its interplay with other cell forces. 相似文献
9.
Bernhard Wallmeyer Sarah Trinschek Sargon Yigit Uwe Thiele Timo Betz 《Biophysical journal》2018,114(1):213-222
Collective cell migration is a fundamental process during embryogenesis and its initial occurrence, called epiboly, is an excellent in vivo model to study the physical processes involved in collective cell movements that are key to understanding organ formation, cancer invasion, and wound healing. In zebrafish, epiboly starts with a cluster of cells at one pole of the spherical embryo. These cells are actively spreading in a continuous movement toward its other pole until they fully cover the yolk. Inspired by the physics of wetting, we determine the contact angle between the cells and the yolk during epiboly. By choosing a wetting approach, the relevant scale for this investigation is the tissue level, which is in contrast to other recent work. Similar to the case of a liquid drop on a surface, one observes three interfaces that carry mechanical tension. Assuming that interfacial force balance holds during the quasi-static spreading process, we employ the physics of wetting to predict the temporal change of the contact angle. Although the experimental values vary dramatically, the model allows us to rescale all measured contact-angle dynamics onto a single master curve explaining the collective cell movement. Thus, we describe the fundamental and complex developmental mechanism at the onset of embryogenesis by only three main parameters: the offset tension strength, α, that gives the strength of interfacial tension compared to other force-generating mechanisms; the tension ratio, δ, between the different interfaces; and the rate of tension variation, λ, which determines the timescale of the whole process. 相似文献
10.
Beth A. Holloway Sol Gomez de la Torre Canny Ying Ye Diane C. Slusarski Christina M. Freisinger Roland Dosch Margaret M. Chou Daniel S. Wagner Mary C. Mullins 《PLoS genetics》2009,5(3)
One of the earliest morphogenetic processes in the development of many animals is epiboly. In the zebrafish, epiboly ensues when the animally localized blastoderm cells spread, thin over, and enclose the vegetally localized yolk. Only a few factors are known to function in this fundamental process. We identified a maternal-effect mutant, betty boop (bbp), which displays a novel defect in epiboly, wherein the blastoderm margin constricts dramatically, precisely when half of the yolk cell is covered by the blastoderm, causing the yolk cell to burst. Whole-blastoderm transplants and mRNA microinjection rescue demonstrate that Bbp functions in the yolk cell to regulate epiboly. We positionally cloned the maternal-effect bbp mutant gene and identified it as the zebrafish homolog of the serine-threonine kinase Mitogen Activated Protein Kinase Activated Protein Kinase 2, or MAPKAPK2, which was not previously known to function in embryonic development. We show that the regulation of MAPKAPK2 is conserved and p38 MAP kinase functions upstream of MAPKAPK2 in regulating epiboly in the zebrafish embryo. Dramatic alterations in calcium dynamics, together with the massive marginal constrictive force observed in bbp mutants, indicate precocious constriction of an F-actin network within the yolk cell, which first forms at 50% epiboly and regulates epiboly progression. We show that MAPKAPK2 activity and its regulator p38 MAPK function in the yolk cell to regulate the process of epiboly, identifying a new pathway regulating this cell movement process. We postulate that a p38 MAPKAPK2 kinase cascade modulates the activity of F-actin at the yolk cell margin circumference allowing the gradual closure of the blastopore as epiboly progresses. 相似文献
11.
Cell surface changes occurring before and during gastrulation in Xenopus laevis embryos have been examined by scanning electron microscopy (SEM). Our study covers the period of development from very young blastulae (stage 7) to late gastrulae (stage . Before the onset of the epibolic movement there is evidence of locomotory activity of the cells lining the blastocoel at the animal pole. In the medim- (stage 8) and small-cell (stage 9) blastula, when pregastrulation movements are progressing rapidly, microvilli appear in the interstices between cells, both at the animal and at the vegetal pole. In the gastrula, most of the cells close to the blastopore have either their entire exposed surface or part of it covered with microvilli. On the other hand, the cells that have just reached the blastopore and have become clubshaped do not display microvilli on their surfaces; microvilli are also absent on the surface of the cells that have undergone invagination. The invaginated chorda-mesoderm is made up of single fibroblastlike cells with long thin filopodia which are interwoven with those of nearby cells. The observations are discussed in relation to changes in cell-to-cell connections and to the role of cell surface organization in the morphogenetic movements of gastrulation. 相似文献
12.
Fibronectin (FN) is reported to be important for early morphogenetic movements in a variety of vertebrate embryos, but the cellular basis for this requirement is unclear. We have used confocal and digital time-lapse microscopy to analyze cell behaviors in Xenopus gastrulae injected with monoclonal antibodies directed against the central cell-binding domain of fibronectin. Among the defects observed is a disruption of fibronectin matrix assembly, resulting in a failure of radial intercalation movements, which are required for blastocoel roof thinning and epiboly. We identified two phases of FN-dependent cellular rearrangements in the blastocoel roof. The first involves maintenance of early roof thinning in the animal cap, and the second is required for the initiation of radial intercalation movements in the marginal zone. A novel explant system was used to establish that radial intercalation in the blastocoel roof requires integrin-dependent contact of deep cells with fibronectin. Deep cell adhesion to fibronectin is sufficient to initiate intercalation behavior in cell layers some distance from the substrate. Expression of a dominant-negative beta1 integrin construct in embryos results in localized depletion of the fibronectin matrix and thickening of the blastocoel roof. Lack of fibronectin fibrils in vivo is correlated with blastocoel roof thickening and a loss of deep cell polarity. The integrin-dependent binding of deep cells to fibronectin is sufficient to drive membrane localization of Dishevelled-GFP, suggesting that a convergence of integrin and Wnt signaling pathways acts to regulate radial intercalation in Xenopus embryos. 相似文献
13.
14.
During epiboly stages the cells (called deep blastomeres) which will form the definitive embryo disperse over the surface of the yolk sphere, only later aggregating and developing an embryonic axis. Five different statistical tests were used to study the pattern formed by the deep blastomeres during epiboly and early dispersed stages. The two most reliable tests, based on the distance from each deep blastomere within a selected area to its nearest neighboring cell, indicate that the distribution pattern changes from regular during epiboly stages to random during dispersed stages 1 and 2. Careful observation and time-lapse microphotography revealed some aspects of how the cells set up the regular pattern. The deep blastomeres exhibit a variety of cell extensions, with which they often contact one another. When two deep blastomeres make contact during epiboly stages, they soon break the contact and move apart; they overlap one another only rarely. Deep blastomeres are frequently located at, and are even elongated along, borders of the overlying flat cells (enveloping layer cells). These two mechanisms, one similar to contact inhibition of cell movement, the other to contact guidance, may contribute to the rather regular spacing of the deep blastomeres as well as to their arrangement in rows during epiboly stages. 相似文献
15.
Spatial distribution and orientation of microfolds arising during invagination of the outer layer of suprablastoporal zone into the blastopore dorsal lip and changes of the lip shape were studied in Rana ridibunda embryos using statistical analysis of a normal individual variability. Active invagination of the cells into the lip correlated with deviation of the orientation of microfolds from the normal in the points of their intersection with the zone of dorsal lip inflection and their orientation is normalized upon transition of the cells across the inflection zone. Frequency distribution of the angle of microfold deviation from the normal is close to the exponential and, therefore, the angle of deviation is an analog of the potential energy of cells-components of the microfold: the bigger the deviation angle, the higher the potential energy. The minimum potential energy is observed at the normal orientation of microfolds, i.e., when it coincides with the radius of the dorsal lip curvature at the point of intersection with the microfold. The following mechanism of dorsal lip formation has been proposed: equatorial contraction of cells upon their invagination into the dorsal lip causes deviation of cell flux orientation from the normal orientation and the normal orientation is restored through an increase in the local curvature of dorsal lip. When the orientation of cell fluxes is normalized, invagination of cells in the dorsal lip ceases. The wave of normalization overtakes the wave of cell invagination into the dorsal lip at the lip angle length 120 degrees. At this moment, the archenteron roof is mechanically detached from the superficial cells of the suprablastoporal zone and lateral blastopore lips and this determines separation of the presumptive notochord. 相似文献
16.
Kiriukhin DO Shustikova LA Kopantseva EE Luchinskaia NN Beliavskiĭ AV 《Biofizika》2010,55(6):1002-1007
The development of multicellular organisms is a complicated coordinated process of the movement of groups of embryonic cells, which is controlled by many regulatory systems. At present little is known about the regulation of the earliest manifestations of the movement in the embryogenesis: epiboly and radial intercalation. The coordinators of these processes may be small GTPases of the Rho family and their activators, the factors of exchange of guanylic nucleotides. It has been shown in this work that the overexpression of the factor of exchange of guanylic nucleotides xLARG in Xenopus laevis embryos leads to an increase in the amount of the active form of xLARG. In addition, an increase in the expression of xLARG disturbs the process of radial intercalation. The data obtained suggest that xLARG is involved in maintaining the xLARG activation level necessary for the occurrence of epiboly. 相似文献
17.
Formation of germ layers and roles of the dorsal lip of the blastopore in normally developing embryos of the newt Cynops pyrrhogaster 总被引:3,自引:0,他引:3
H Imoh 《The Journal of experimental zoology》1988,246(3):258-270
Three-dimensional relationships between tissues during the formation of germ layers were studied in sections of normally developing embryos of the newt, Cynops pyrrhogaster. In gastrulae, the inner postinvolution layer was not in direct contact with the outer preinvolution layer as a result of the presence of an intervening layer of cells. Only after the formation of the yolk plug, a narrow strip of primitive notochord, which consisted of columnar cells, established a close contact with the central part of the overlaying presumptive neural plate. The primitive notochord was also linked to endoderm at its right and left margins, facing the archenteron. Mesodermal cells other than notochord cells were mesenchymal until the neurula stage, when primitive somites appeared on both sides of the notochord. From a comparison of the relative locations of tissues in embryos at different stages of development, it was shown that the notochord elongates by a remodeling of the mass of the primitive notochord, and that, as the anteriorly directed translocation of the neural area and the invagination of endoderm occur, these processes keep pace with the elongation of the notochord. These observations suggest organizing or guiding roles for the notochord in the formation of germ layers. A role for the dorsal lip of the blastopore as the organizer is discussed in relation to the origin of the notochord. 相似文献
18.
This paper summarizes evidence for the following picture ofFundulus epiboly, with an eye toward laying groundwork for futureinvestigation. The major force in epiboly is the yolk syncytiallayer (YSL). Prior to epiboly, it spreads well beyond the borderof the blastoderm to form the wide external YSL (E-YSL). Thishas contractile properties, which, however, are restrained priorto epiboly by the attached enveloping layer (EVL) of the blastoderm.Epiboly begins when the E-YSL contracts and narrows, throwingits surface into folds and pulling the internal YSL (I-YSL)and the attached EVL vegetally. When the narrowing of the E-YSLhas ceased, it is postulated that its contractility continuesas a circumferential wave of vegetally directed contractionthat moves over the yolk toward the vegetal pole, dragging theI-YSL and the attached EVL (and blastoderm) with it. The mostobvious visible manifestation of this wave is a marked marginalconstriction, where the YSL joins the yolk cytoplasmic layer(YCL). As this contractile wave passes over the yolk, cytoplasmfrom the YCL mingles with that of the advancing E-YSL, and YCLsurface adds to the already highly convoluted surface of theE-YSL. This folded surface is the site of a thin, highly localizedband of rapid endocytosis that encircles the egg and passesover it with the E-YSL in a wave throughout epiboly. This internalization,which is receptor independent and therefore somehow programmed,accompanies the putative contractile wave, and accounts forthe disappearance of the surface of the YCL. Since the YCL surfacestands in the way of the advancing YSL, its internalizationis part of the mechanism of epiboly. As the I-YSL expands inresponse to this marginal pull, its abundant microvilli graduallydisappear, providing surface for its epiboly. The firmly attachedEVL likewise expands toward the vegetal pole in response tothe pull of the autonomously expanding YSL. As epiboly of theEVL progresses, it adjusts to the geometric problems posed bya sheet expanding over a sphere by active cell rearrangementwithin the cell monolayer. Thus, epiboly of the EVL has an activeas well as a passive component. Deep cells are not causallyinvolved in epiboly, but move about in coordinated ways in theconstantly increasing space between the I-YSL and the EVL providedby epiboly and form the germ ring and the embryonic shield andeventually the embryo proper. An attempt is made to pull allof this together, and more, in order to achieve as comprehensivean understanding of epiboly as present evidence will allow. 相似文献
19.
R. E. Keller 《Journal of morphology》1978,157(2):223-247
Time-lapse cinemicrography was used to show what changes in the number, size, shape, arrangement and what movements of apices of superficial cells occur during epiboly, extension, convergence and blastopore formation in the blastula or gastrula of Xenopus laevis. Epiboly of the animal region occurs by apical expansion of superficial cells at a nearly constant rate from the midblastula to the midgastrula stage. Egression of deep cells into the superficial layer does not occur. Extension of the dorsal marginal zone begins in the late blastula stage with the rapid spreading of the apices of cells in this region and this continues until the onset of neurulation when rapid shrinkage begins. Extension and convergence of the dorsal marginal zone occurs by a rearrangement in which individual cells exchange neighbors and by a change in the shape of the cell apices. Regional differences in apical expansion are accompanied by differences in rate of anticlinal division of superficial cells such that cells in all sectors of the animal region and the marginal zone show similar patterns of decrease in apparent apical area. Shrinkage of the apices of bottle cells during blastopore formation is described. From this and other studies, a model of the cellular behavior of epiboly, extension and convergence is constructed and several hypotheses as to how these activities might generate the mechanical forces of the gastrulation movements are presented. 相似文献
20.
Solnica-Krezel L 《Current biology : CB》2005,15(6):R213-R228
Vertebrate embryogenesis entails an exquisitely coordinated combination of cell proliferation, fate specification and movement. After induction of the germ layers, the blastula is transformed by gastrulation movements into a multilayered embryo with head, trunk and tail rudiments. Gastrulation is heralded by formation of a blastopore, an opening in the blastula. The axial side of the blastopore is marked by the organizer, a signaling center that patterns the germ layers and regulates gastrulation movements. During internalization, endoderm and mesoderm cells move via the blastopore beneath the ectoderm. Epiboly movements expand and thin the nascent germ layers. Convergence movements narrow the germ layers from lateral to medial while extension movements elongate them from head to tail. Despite different morphology, parallels emerge with respect to the cellular and genetic mechanisms of gastrulation in different vertebrate groups. Patterns of gastrulation cell movements relative to the blastopore and the organizer are similar from fish to mammals, and conserved molecular pathways mediate gastrulation movements. 相似文献