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1.
Development ; 141(19): 3672-82, 2014 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-25249459

RESUMEN

Morphogenetic processes often involve the rapid rearrangement of cells held together by mutual adhesion. The dynamic nature of this adhesion endows tissues with liquid-like properties, such that large-scale shape changes appear as tissue flows. Generally, the resistance to flow (tissue viscosity) is expected to depend on the cohesion of a tissue (how strongly its cells adhere to each other), but the exact relationship between these parameters is not known. Here, we analyse the link between cohesion and viscosity to uncover basic mechanical principles of cell rearrangement. We show that for vertebrate and invertebrate tissues, viscosity varies in proportion to cohesion over a 200-fold range of values. We demonstrate that this proportionality is predicted by a cell-based model of tissue viscosity. To do so, we analyse cell adhesion in Xenopus embryonic tissues and determine a number of parameters, including tissue surface tension (as a measure of cohesion), cell contact fluctuation and cortical tension. In the tissues studied, the ratio of surface tension to viscosity, which has the dimension of a velocity, is 1.8 µm/min. This characteristic velocity reflects the rate of cell-cell boundary contraction during rearrangement, and sets a limit to rearrangement rates. Moreover, we propose that, in these tissues, cell movement is maximally efficient. Our approach to cell rearrangement mechanics links adhesion to the resistance of a tissue to plastic deformation, identifies the characteristic velocity of the process, and provides a basis for the comparison of tissues with mechanical properties that may vary by orders of magnitude.


Asunto(s)
Adhesión Celular/fisiología , Movimiento Celular/fisiología , Embrión no Mamífero/citología , Modelos Biológicos , Morfogénesis/fisiología , Xenopus/embriología , Animales , Ficoll , Procesamiento de Imagen Asistido por Computador , Microscopía Confocal , Tensión Superficial , Viscosidad
2.
Development ; 141(19): 3649-61, 2014 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-25209247

RESUMEN

Xenopus provides a well-studied model of vertebrate gastrulation, but a central feature, the movement of the mesoderm to the interior of the embryo, has received little attention. Here, we analyze mesoderm involution at the Xenopus dorsal blastopore lip. We show that a phase of rapid involution - peak involution - is intimately linked to an early stage of convergent extension, which involves differential cell migration in the prechordal mesoderm and a new movement of the chordamesoderm, radial convergence. The latter process depends on Xenopus Brachyury, the expression of which at the time of peak involution is controlled by signaling through the ephrin receptor, EphA4, its ligand ephrinB2 and its downstream effector p21-activated kinase. Our findings support a conserved role for Brachyury in blastopore morphogenesis.


Asunto(s)
Proteínas Fetales/metabolismo , Gástrula/embriología , Regulación del Desarrollo de la Expresión Génica/fisiología , Mesodermo/embriología , Receptor EphA4/metabolismo , Transducción de Señal/fisiología , Proteínas de Dominio T Box/metabolismo , Xenopus/embriología , Animales , Movimiento Celular/fisiología , Regulación del Desarrollo de la Expresión Génica/genética , Procesamiento de Imagen Asistido por Computador , Hibridación in Situ , Microscopía Electrónica de Rastreo , Morfolinos/genética , Transducción de Señal/genética
3.
Elife ; 112022 04 11.
Artículo en Inglés | MEDLINE | ID: mdl-35404236

RESUMEN

The morphogenic process of convergent thickening (CT) was originally described as the mediolateral convergence and radial thickening of the explanted ventral involuting marginal zone (IMZ) of Xenopus gastrulae (Keller and Danilchik, 1988). Here, we show that CT is expressed in all sectors of the pre-involution IMZ, which transitions to expressing convergent extension (CE) after involution. CT occurs without CE and drives symmetric blastopore closure in ventralized embryos. Assays of tissue affinity and tissue surface tension measurements suggest CT is driven by increased interfacial tension between the deep IMZ and the overlying epithelium. The resulting minimization of deep IMZ surface area drives a tendency to shorten the mediolateral (circumblastoporal) aspect of the IMZ, thereby generating tensile force contributing to blastopore closure (Shook et al., 2018). These results establish CT as an independent force-generating process of evolutionary significance and provide the first clear example of an oriented, tensile force generated by an isotropic, Holtfreterian/Steinbergian tissue affinity change.


Asunto(s)
Evolución Biológica , Gástrula , Animales , Movimiento Celular , Morfogénesis , Xenopus laevis
4.
J Cell Biol ; 208(6): 839-56, 2015 Mar 16.
Artículo en Inglés | MEDLINE | ID: mdl-25778923

RESUMEN

Cleft-like boundaries represent a type of cell sorting boundary characterized by the presence of a physical gap between tissues. We studied the cleft-like ectoderm-mesoderm boundary in Xenopus laevis and zebrafish gastrulae. We identified the transcription factor Snail1 as being essential for tissue separation, showed that its expression in the mesoderm depends on noncanonical Wnt signaling, and demonstrated that it enables paraxial protocadherin (PAPC) to promote tissue separation through two novel functions. First, PAPC attenuates planar cell polarity signaling at the ectoderm-mesoderm boundary to lower cell adhesion and facilitate cleft formation. Second, PAPC controls formation of a distinct type of adhesive contact between mesoderm and ectoderm cells that shows properties of a cleft-like boundary at the single-cell level. It consists of short stretches of adherens junction-like contacts inserted between intermediate-sized contacts and large intercellular gaps. These roles of PAPC constitute a self/non-self-recognition mechanism that determines the site of boundary formation at the interface between PAPC-expressing and -nonexpressing cells.


Asunto(s)
Cadherinas/fisiología , Factores de Transcripción/fisiología , Proteínas de Xenopus/fisiología , Actinas/metabolismo , Animales , Tipificación del Cuerpo , Adhesión Celular , Polaridad Celular , Gástrula/embriología , Gástrula/metabolismo , Mesodermo/citología , Mesodermo/metabolismo , Protocadherinas , Receptores Acoplados a Proteínas G/metabolismo , Proteínas de Xenopus/metabolismo , Xenopus laevis , Pez Cebra , Proteínas de Pez Cebra/fisiología
5.
Cell Signal ; 24(12): 2337-48, 2012 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-22884502

RESUMEN

Primitive endoderm formation from the inner cell mass is one of the earliest known cell fate decisions made in the mouse embryo. The mechanisms involved in orchestrating this process are not fully understood and are difficult to study in vivo. The F9 teratocarcinoma cell line is an in vitro model used to circumvent many technical problems surrounding the study of extraembryonic endoderm differentiation. F9 cells treated with retinoic acid differentiate to primitive endoderm and this is accompanied by the activation of canonical Wnt-ß-catenin signalling. Reactive oxygen species can modulate this signalling pathway, but whether they are sufficient to induce extraembryonic endoderm in vitro is not known. In the present study, a sustained increase in ROS levels was found in retinoic acid-treated F9 cells. An increase in Tcf-Lef transcriptional activity, a read out of Wnt-ß-catenin signalling, was also seen in response to exogenous H(2)O(2). Analysis from immunoblots, immunocytochemistry and real time PCR revealed the presence of markers of differentiation and a reduction in the expression of a marker of proliferation, confirming that H(2)O(2)-treated F9 cells developed into primitive endoderm. In contrast, exposing retinoic acid-treated cells to antioxidants impeded differentiation. Real time PCR was also used to identify candidates responsible for the observed elevation in ROS production. Results indicated that the NADPH oxidase 1, 2, 3 and 4 and Duox2 genes were RA responsive. Furthermore, the NADPH oxidase inhibitor, diphenyleneiodonium chloride was shown to attenuate primitive endoderm formation. Together, these results shed new light on how early mouse embryogenesis might be influenced by the crosstalk involving ROS and the Wnt-ß-catenin signalling pathway.


Asunto(s)
Endodermo/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Proteínas Wnt/metabolismo , Animales , Antioxidantes/farmacología , Diferenciación Celular/efectos de los fármacos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Oxidasas Duales , Endodermo/citología , Endodermo/efectos de los fármacos , Peróxido de Hidrógeno/farmacología , Ratones , NADPH Oxidasas/antagonistas & inhibidores , NADPH Oxidasas/genética , NADPH Oxidasas/metabolismo , Compuestos Onio/farmacología , Transducción de Señal/efectos de los fármacos , Factores de Transcripción TCF/genética , Factores de Transcripción TCF/metabolismo , Transcripción Genética , Tretinoina/farmacología , beta Catenina/metabolismo
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