Investigating the Neural Crest Cell Migration in Shark Embryos using Fox D3 and Acetylated Tubulin
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Since the development of the peripheral nervous system (PNS) is a critical component of vertebrate development, exploring the processes involved in PNS development is important. A part of PNS development is the migration of trunk neural crest cells (NCC) since the trunk NCC differentiate into cell lineages needed for the PNS. Therefore, this study chose to observe NCC migration since it will provide insights on PNS development. This study examines NCC migration in shark embryos through three facets: when the stage migration begins and ends, when migration is at its peak, and the different pathways that can be observed. In order to track NCC, shark embryos ranging from stage 20 to 30, were cut into thin sections using a vibratome machine. The sections were then stained with the primary antibodies: Fox d3 and Acetylated Tubulin and the matching secondary antibodies. In addition to taking pictures of sections of the embryos, images of the whole embryo and images of DiI staining was collected. DiI labeling and images of a stage 20 shark revealed migration begins as early as stage 19-20 with the primary pathway being dorsolateral. Within the dorsolateral pathway, cells migrated in clusters – collective cell migration. Whole mount images demonstrated the highest level of delamination occurred in stage 25 sharks. Therefore, stage 25 could be considered the peak of migration with cells transitioning from cluster formation to direct streams as dorsal root ganglion (DRGs) are beginning to form. Stage 30 can be the decline or end of migration and by that stage, elements of dorsolateral, ventromedial, and ventrolateral pathway is observed. Fox d3’s presence in all stages allowed these conclusions to be drawn and hence it can be considered a strong NCC marker for stages 20-30. Overall, these results provide a clearer picture on neural crest and embryonic development of sharks, specifically NCC migration which can lead to future projects that investigate specific components of migration.