Live Imaging of the Dynamics of Mammalian Neural Crest Cell Migration

Neural crest cells (NCC) are a multipotent cell population that play an important role in vertebrate development. Often touted as the fourth-germ layer, NCC are induced at the border of the neural and non-neural ectoderm during the neurulation phase of embryogenesis. NCC undergo an epithelial to mesenchymal transition (EMT) that facilitates their delamination and migration throughout the embryo. After reaching their final destination, NCC then differentiate into numerous distinct cell types including neurons and glia, melanocytes, and craniofacial chondrocytes and osteoblasts. Research into the signals and mechanisms regulating each step of NCC development has been instrumental to our understanding of vertebrate development, evolution, and disease. However, studying the single and collective cellular dynamics of mammalian NCC migration has proven difficult due to the challenges accessing, and limitations visualizing, NCC within an embryo that develops in utero. The following chapter describes methods for studying the dynamics of cranial NCC migration in whole mouse embryos and in two-dimensional (2D) and 3D explant cultures of the neural plate, but these methods can be adapted for NCC at any axial level of the embryo.