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Abstract
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.
Highlights
During vertebrate embryonic development, morphogenetic processes shape a multilayered three-dimensioned embryo consisting of tissues derived from ectoderm, endoderm, and mesoderm [1]
The development of the nervous system is a typical illustration of the importance of this tissue interaction, whereby the brain and the spinal cord arise from dorsal ectoderm via induction by molecular signals derived from the underlying dorsal mesoderm [3]
Saraiva contributed with all other contributors. Demonstrating this point involved the use of a refined mechano-molecular toolbox aimed to determine the impact that modifying the activity of planar cell polarity (PCP) in the mesoderm has on both mesoderm cell migration and stiffness, as well as on Neural crest (NC) polarity and migration
Summary
Morphogenetic processes shape a multilayered three-dimensioned embryo consisting of tissues derived from ectoderm, endoderm, and mesoderm [1]. The development of the nervous system is a typical illustration of the importance of this tissue interaction, whereby the brain and the spinal cord arise from dorsal ectoderm via induction by molecular signals derived from the underlying dorsal mesoderm (known by amphibian embryologists as the “Spemann’s organizer”) [3]. Demonstrating this point involved the use of a refined mechano-molecular toolbox aimed to determine the impact that modifying the activity of PCP in the mesoderm has on both mesoderm cell migration and stiffness, as well as on NCs polarity and migration. Our work is organized in several sections, where we will explain how to target dorsal mesoderm in order to molecularly modify PCP activity in this tissue (Subheading 3.1); how to determine the impact of PCP inhibition
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