Mechanisms of Morphogenesis Lab, Gulbenkian Institute of Science (IGC), Oeiras, Portugal.
Methods Mol Biol. 2022;2438:495-515. doi: 10.1007/978-1-0716-2035-9_29.
During vertebrate embryogenesis, tissues interact and influence each other's development to shape an embryo. While communication by molecular components has been extensively explored, the role of mechanical interaction between tissues during embryogenesis is just starting to be revealed. Addressing mechanical involvement in morphogenesis has traditionally been challenging mainly due to the lack of proper tools to measure and modify mechanical environments of cells in vivo. We have recently used atomic force microscopy (AFM) to show that the migration of the Xenopus laevis cephalic neural crest cells is triggered by stiffening of the mesoderm, a tissue that neural crest cells use as a migratory substrate in vivo. Interestingly we showed that the activity of the planar cell polarity (PCP) pathway is required to mediate this novel mechanical interaction between two tissues. In this chapter, we share the toolbox that we developed to study the role of PCP signaling in mesoderm cell accumulation and stiffening (in vivo) as well as the impact of mesoderm stiffness in promoting neural crest cell polarity and migration (ex vivo). We believe that these tools can be of general use for investigators interested in addressing the role of mechanical inputs in vivo and ex vivo.
在脊椎动物胚胎发生过程中,组织相互作用并影响彼此的发育,从而形成胚胎。虽然分子成分的交流已经被广泛研究,但组织之间机械相互作用在胚胎发生中的作用才刚刚开始被揭示。由于缺乏适当的工具来测量和改变细胞在体内的机械环境,传统上解决机械参与形态发生一直具有挑战性。我们最近使用原子力显微镜(AFM)表明,非洲爪蟾的头颈部神经嵴细胞的迁移是由中胚层的变硬引发的,神经嵴细胞在体内将中胚层用作迁移底物。有趣的是,我们表明平面细胞极性(PCP)途径的活性对于介导两种组织之间的这种新的机械相互作用是必需的。在本章中,我们分享了我们开发的工具包,用于研究 PCP 信号在中胚层细胞积累和变硬(体内)中的作用,以及中胚层硬度在促进神经嵴细胞极性和迁移(体外)中的作用。我们相信,对于有兴趣在体内和体外解决机械输入作用的研究人员来说,这些工具具有普遍的用途。