Date Thesis Awarded

4-2019

Document Type

Honors Thesis

Degree Name

Bachelors of Science (BS)

Department

Biology

Advisor

Margaret Saha, Ph. D.

Committee Members

Jennifer Bestman, Ph. D.

Paul Heideman, Ph. D.

Robin Looft-Wilson, Ph. D.

Abstract

Neural development is a highly regulated process that requires coordinated signaling in order to develop a properly functioning nervous system. One of these key signaling pathways is the Notch pathway, a highly conserved signaling pathway that plays a key role in development, particularly neural development. Recent studies have linked Ttyh1, a volume-sensitive chloride channel with a calcium-binding site, to the Notch signaling pathway. However, despite its importance in development and disease, Ttyh1 remains poorly studied. Previous work in the lab has shown that Ttyh1 is highly expressed in the adult and developing nervous system, expression that has implications for the maintenance of neural stem cells and numerous cellular functions, such as cell communication, adhesion, and migration. This thesis project investigated the role of Ttyh1 in neuro-development by genetically knocking it out using CRISPR/Cas9 genomic editing technology and investigating the expression of neural marker genes and the other members of the tweety family. Chromogenic in situ hybridization revealed knockout of Ttyh1 resulted in differential expression of Sox2, a progenitor cell marker, and tubb2b, a differentiated neuronal marker. RT-qPCR analysis of Ttyh1 knockouts did not result in a consistent significant difference in gene expression. These results suggest that Ttyh1 may be involved in neural stem cell development, but further investigation is needed to confirm its precise role.

Available for download on Saturday, May 01, 2021

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