Using zebrafish to elucidate the expression and gene regulatory network of the Genomic screen homeobox transcription factors and bring innovative science learning experiences to West Virginia

Central nervous system (CNS) development requires a code of regionally expressed transcription factors that impart initial neuronal cell identity, connectivity, and function. The absence of a transcription factor code would eliminate the neurodevelopmental logic producing diverse cell types in the CNS. Furthermore, early disruptions in transcription factor expression can affect later connectivity and function of neuronal circuits mediating sensory processing, defects in which are often observed as comorbid with various neurodevelopmental disorders (NDDs). Characterizing transcription factor expression and function is therefore an essential step in discerning the molecular mechanisms underlying human NDDs. genomic screen homeobox 1 and 2 (gsx1 and gsx2) encode homeobox transcription factors expressed in the vertebrate CNS. In mouse, Gsx1 and Gsx2 regulate neuronal progenitor proliferation and specification in developing brain regions such as the telencephalon, cerebellum, and spinal cord. However, many details about their expression and function across other brain regions in vertebrates and in zebrafish specifically remain under investigated. An initial step in elucidating the relationship between Gsx1 and Gsx2, CNS development, and NDD etiology is to first obtain a molecular framework-level understanding of the importance Gsx1 and Gsx2 hold across CNS regions. In this dissertation, I present a comprehensive analysis of gsx1 and gsx2 expression, regulatory relationship, and a putative gene regulatory network in the zebrafish CNS. Additionally, I describe efforts that utilize zebrafish as a hands-on science learning tool in a science-education partnership targeted at high school students from West Virginia that are underrepresented in STEM. I begin this dissertation by discussing and describing the spatiotemporal expression profiles of gsx1 and gsx2 in zebrafish and phenotypes of gsx1 and gsx2 mutant zebrafish. I then show their proposed regulatory relationship to each other in zebrafish and provide a thorough analysis of their putative gene regulatory network through in silico analyses. Next, I present evidence that using zebrafish in a rural science-education partnership fosters increased science confidence in high school students both in-person and virtually. I also show that teachers in West Virginia are interested in using zebrafish for science education, however require varied support based on funding, the availability of classroom supplies, and experimental support. Together, this dissertation highlights how zebrafish sit at the intersection of research and education to contribute to knowledge of genes essential for early neurodevelopment and the continued enhancement of science-education partnerships in West Virginia and beyond.