RESUMO
Collective migration of cohesive tissues is a fundamental process in morphogenesis and is particularly well illustrated during gastrulation by the rapid and massive internalization of the mesoderm, which contrasts with the much more modest movements of the ectoderm. In the Xenopus embryo, the differences in morphogenetic capabilities of ectoderm and mesoderm can be connected to the intrinsic motility of individual cells, very low for ectoderm, high for mesoderm. Surprisingly, we find that these seemingly deep differences can be accounted for simply by differences in Rho-kinases (Rock)-dependent actomyosin contractility. We show that Rock inhibition is sufficient to rapidly unleash motility in the ectoderm and confer it with mesoderm-like properties. In the mesoderm, this motility is dependent on two negative regulators of RhoA, the small GTPase Rnd1 and the RhoGAP Shirin/Dlc2/ArhGAP37. Both are absolutely essential for gastrulation. At the cellular and tissue level, the two regulators show overlapping yet distinct functions. They both contribute to decrease cortical tension and confer motility, but Shirin tends to increase tissue fluidity and stimulate dispersion, while Rnd1 tends to favor more compact collective migration. Thus, each is able to contribute to a specific property of the migratory behavior of the mesoderm. We propose that the "ectoderm to mesoderm transition" is a prototypic case of collective migration driven by a down-regulation of cellular tension, without the need for the complex changes traditionally associated with the epithelial-to-mesenchymal transition.
Assuntos
Actomiosina/metabolismo , Ectoderma/fisiologia , Mesoderma/fisiologia , Animais , Movimento Celular/genética , Regulação para Baixo/fisiologia , Ectoderma/embriologia , Embrião não Mamífero , Transição Epitelial-Mesenquimal/fisiologia , Proteínas Ativadoras de GTPase/genética , Proteínas Ativadoras de GTPase/metabolismo , Gastrulação/fisiologia , Regulação da Expressão Gênica no Desenvolvimento , Mesoderma/embriologia , Morfogênese/fisiologia , Transporte Proteico/genética , Transdução de Sinais/genética , Distribuição Tecidual/genética , Proteínas de Xenopus/genética , Proteínas de Xenopus/metabolismo , Xenopus laevis , Proteínas rho de Ligação ao GTP/genética , Proteínas rho de Ligação ao GTP/metabolismoRESUMO
Recent studies have shown that heterotrimeric G proteins are involved in the regulation of the canonical Wnt/beta-Catenin pathway. However, the mechanism(s) behind this involvement is (are) poorly understood. Our previous results have shown that activation of Galphaq in Xenopus oocytes leads to inhibition of GSK-3beta and stabilization of the beta-Catenin protein, suggesting that Galphaq might stabilize beta-Catenin via inhibition of GSK-3beta. In this study, we have observed similar results in HEK293T cells. In these cells optimal activation of endogenous Galphaq by expressing M3-muscarinic acetylcholine receptor (with or without carbachol treatment), or exposing the cells to thrombin led to an increase of 2 to 3-fold in endogenous cytoplasmic beta-Catenin protein levels. In addition, expression of the activated mutant of Galphaq (GalphaqQL) dramatically enhanced accumulation of exogenous beta-Catenin with no effect on beta-catenin (CTNNB1) gene transcription. The Galphaq-mediated cellular accumulation of beta-Catenin was blocked by expression of a minigene encoding a Galphaq specific inhibitory peptide but not by a minigene encoding a Galphas blocking peptide. Also, expression of GalphaqQL led to a significant reduction in GSK-3beta kinase activity, supporting the idea that the positive role of Galphaq signaling in inducing cellular accumulation of beta-Catenin is mediated through inhibition of GSK-3beta.
Assuntos
Subunidades alfa Gq-G11 de Proteínas de Ligação ao GTP/metabolismo , Quinase 3 da Glicogênio Sintase/metabolismo , Proteínas Wnt/metabolismo , beta Catenina/metabolismo , Linhagem Celular , Subunidades alfa Gq-G11 de Proteínas de Ligação ao GTP/genética , Quinase 3 da Glicogênio Sintase/antagonistas & inibidores , Quinase 3 da Glicogênio Sintase/genética , Glicogênio Sintase Quinase 3 beta , Humanos , Receptor Muscarínico M3/agonistas , Receptor Muscarínico M3/biossíntese , Transcrição Gênica , beta Catenina/genéticaRESUMO
The establishment of sharp boundaries is essential for segregation of embryonic tissues during development, but the underlying mechanism of cell sorting has remained unclear. Opposing hypotheses have been proposed, either based on global tissue adhesive or contractile properties or on local signalling through cell contact cues. Here we use ectoderm-mesoderm separation in Xenopus to directly evaluate the role of these various parameters. We find that ephrin-Eph-based repulsion is very effective at inducing and maintaining separation, whereas differences in adhesion or contractility have surprisingly little impact. Computer simulations support and generalise our experimental results, showing that a high heterotypic interfacial tension between tissues is key to their segregation. We propose a unifying model, in which conditions of sorting previously considered as driven by differential adhesion/tension should be viewed as suboptimal cases of heterotypic interfacial tension.The mechanisms that cause different cells to segregate into distinct tissues are unclear. Here the authors show in Xenopus that formation of a boundary between two tissues is driven by local tension along the interface rather than by global differences in adhesion or cortical contractility.