Date of Award

12-2026

Document Type

Thesis

Degree Name

Master of Science in Biology

Department

Biology

First Reader/Committee Chair

Bournias-Vardiabasis, Nicole

Abstract

Neural crest cells (NCCs) are a transient, highly migratory embryonic stem cell population unique to vertebrates. During development, they give rise to diverse tissues, including craniofacial cartilage, peripheral neurons, glia, melanocytes, and components of the cardiac outflow tract. As NCCs migrate through complex environments, precise spatial and temporal regulation of gene expression is needed to coordinate migration and developmental fate. Defects in NCC development and migration can result in a broad class of disorders known as neurocristopathies (NCPs), including craniofacial abnormalities, Hirschsprung’s disease, and neuroblastoma. Despite the importance of NCC migration during development, many of the molecular mechanisms regulating this process are still not fully understood.

One mechanism that enables spatial regulation of gene expression is RNA localization, the transport of messenger RNAs (mRNAs) to specific subcellular regions. Previous work, including studies from our lab, demonstrated that in other migratory cell types, specific mRNAs localize to cellular protrusions and contribute to efficient migration. Interestingly, in these systems, disruption of mRNA localization alone is enough to reduce migratory efficiency, suggesting that RNA localization itself plays an important role in regulating migration. However, despite the highly migratory nature of NCCs, RNA localization within migrating neural crest cells has not yet been directly examined.

In this study, neural fold explants isolated from mouse embryos were cultured under different media conditions and embryonic stages to evaluate their ability to generate NCC cellular outgrowths. Cellular identity was assessed using RNA sequencing and immunofluorescence. Overall, E9.5 cultures showed more consistent neural crest characteristics than E8.5 cultures. Single-molecule fluorescence in situ hybridization (smFISH) was then used to examine migration-associated RNAs, including Kif1C. However, technical limitations encountered when combining smFISH with SOX10 immunofluorescence prevented assessment of RNA localization specifically within NCCs. The findings establish a foundation for NCC culture and labeling methods to allow for future investigation of RNA localization during NCC migration.

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