An Analysis of Avian Trunk Neural Crest Cell Migration In Vitro: Investigating the Role and Arrangement of Actin in Migrating Neural Crest Cells

Abstract

Neural crest (NC) cells are multipotent and highly migratory cells that are released from the crest of the neural folds in vertebrates. The NC cells give rise to a plethora of cells and tissues in the developing embryo and adult. The migratory behaviour of individual NC cells is said to be facilitated by actin filaments. Using an in vitro model, avian (quail) trunk neural tubes were cultured to obtain NC cells, in which the migration of individual NC cells could be studied. To determine the role of actin in the migration of NC cells, cytochalasin D was added to some of the cultures in order to block actin polymerisation. Cytochalasin D treated and untreated (control) neural crest cells were imaged over time. An anti-HNK antibody was used to specially detect NC cells in fixed cultures while Phalloidin-Atto590 was used to stain actin. Under control conditions, NC cells exhibited their typical stellate shape and displayed dynamic responses to signals from their external environment (i.e. cell-cell contact) which influenced the morphology of their projections and directed migration. In the cytochalasin D treated cells, NC cell morphology was altered, and the migratory capacity of the cells was significantly reduced. However, cytochalasin D treated cells, retained their ability to respond to environmental cues such as cell-cell contact. Thus, even when actin is disrupted, NC cell migration and their response to their environment appears to be somewhat maintained. These insights highlight the need to explore the molecular basis of NC cell migration especially under disrupted conditions, to further our understanding of neurocristopathies that arise as a result of abnormal NC cell migration.