Honors College Senior Thesis Presentations
Characterizing the role of Glis3 in zebrafish development using whole-mount in situ hybridization (WISH).
Academic Level at Time of Presentation
Senior
Major
Biomedical Sciences
Minor
Chemistry
List all Project Mentors & Advisor(s)
Dr. Gary ZeRuth
Presentation Format
Oral Presentation
Abstract/Description
Zebrafish (Danio rerio) is a frequently used model for studying development sharing many signal transduction pathways with humans. With short generation time, easily analyzable embryos, and genetic malleability, several techniques can be used to study the development, specifically understanding cell differentiation and specification. Focusing on the pancreas as a diabetes model, zebrafish regulate glucose homeostasis in much the same fashion as humans. They develop a principle and secondary islets while producing all the same endocrine and exocrine cell types. Despite this homology, zebrafish pancreatic development does seem to differ from mammals when it comes to transcriptional regulation and cell fate specification. One specific transcription factor has been implicated in the pancreas development in synergism with other transcription factors known in pancreas development. This protein is Glis3, a Kruppel-like zinc finger protein that transcriptionally regulates several target genes as both an activator and repressor of gene expression. Preliminary evidence has shown that, in the zebrafish, the removal of Glis3 correlates to changes in insulin expression along with phenotypic changes in development that include the pancreas, and kidneys among other organs. The aims of this thesis are to investigate and characterize such phenotypic changes from a CRISPR/Cas9 knockdown of the Glis3 gene. This knockdown model, through RT-PCR and whole-mount in situ hybridization, will allow us to better characterize the role of Glis3 in zebrafish development, particularly in relation to turning on various known transcription factors such as ascl1b and ptf1a (both key regulators in mammalian and zebrafish pancreas development).
Spring Scholars Week 2022 Event
Honors College Senior Thesis Presentations
Characterizing the role of Glis3 in zebrafish development using whole-mount in situ hybridization (WISH).
Zebrafish (Danio rerio) is a frequently used model for studying development sharing many signal transduction pathways with humans. With short generation time, easily analyzable embryos, and genetic malleability, several techniques can be used to study the development, specifically understanding cell differentiation and specification. Focusing on the pancreas as a diabetes model, zebrafish regulate glucose homeostasis in much the same fashion as humans. They develop a principle and secondary islets while producing all the same endocrine and exocrine cell types. Despite this homology, zebrafish pancreatic development does seem to differ from mammals when it comes to transcriptional regulation and cell fate specification. One specific transcription factor has been implicated in the pancreas development in synergism with other transcription factors known in pancreas development. This protein is Glis3, a Kruppel-like zinc finger protein that transcriptionally regulates several target genes as both an activator and repressor of gene expression. Preliminary evidence has shown that, in the zebrafish, the removal of Glis3 correlates to changes in insulin expression along with phenotypic changes in development that include the pancreas, and kidneys among other organs. The aims of this thesis are to investigate and characterize such phenotypic changes from a CRISPR/Cas9 knockdown of the Glis3 gene. This knockdown model, through RT-PCR and whole-mount in situ hybridization, will allow us to better characterize the role of Glis3 in zebrafish development, particularly in relation to turning on various known transcription factors such as ascl1b and ptf1a (both key regulators in mammalian and zebrafish pancreas development).