Investigation of the Role of Twist2 on Bone Development in Zebrafish
Sciences and Mathematics, College of
Biology, Department of
BURS Faculty Advisor
Cell fate determination during embryogenesis impacts the proper development of mature organ systems in eukaryotic organisms. Following gastrulation, the mesodermal germ layer has a sub-compartment region known as the sclerotome, the cells of which eventually differentiate into bones, ligaments, and the myotendinous elements of the axial skeleton (Brent et. al 2003). In zebrafish, the twist1b and twist2 genes are suggested to play a role in the differentiation of sclerotome cells into these different elements of the axial skeleton (Germanguz et. al 2007). However, the specific mechanisms for how and when this differentiation occurs is largely unknown. To investigate the role of the twist genes in embryonic bone development, we have now used morpholino injections to knock down the twist2 gene in zebrafish embryos, and in situ hybridization to analyze the expression level changes of genetic markers (sox9, nkx3.2) associated with bone formation. We hope that this will further elucidate the role of twist2 in sclerotome differentiation into bone and provide potential insight into the genetic basis of human musculoskeletal diseases that are associated with orthologous TWIST genes in humans. Furthermore, we also used molecular cloning to synthesize novel mRNA probes to target bone markers sox5 and sox6, which we predict to be between sox9 and nkx3.2 in the development of bone from sclerotome.
Jashim, Elma and Glenn, Nikki, "Investigation of the Role of Twist2 on Bone Development in Zebrafish" (2022). Belmont University Research Symposium (BURS). 130.