Date of Award

Summer 8-15-2026

Degree Type

Thesis

Degree Name

MS Biology

Department

Biology

Advisor

C.J. Urso, Ph.D

Committee Member

Edward Tall, Ph.D

Committee Member

Katherine Varandas, Ph.D

Committee Member

C.J. Urso, Ph.D.

Keywords

SH-SY5Y, retinoic acid, neuronal differentiation, zDHHC, RT-qPCR, gene expression

Abstract

Protein S-Palmitoylation is a reversible post-translational modification that regulates protein trafficking, membrane localization, and neuronal function through the activity of the zinc finger Asp-His-His-Cys (zDHHC) family of palmitoyl acyltransferases. Although SH-SY5Y neuroblastoma cells are widely used as an in vitro model of neuronal differentiation, the expression profile of the complete zDHHC family during retinoic acid (RA)-induced differentiation has not been systematically characterized. The objective of this study was to characterize the expression of all 23 human zDHHC genes during RA induced differentiation of SH-SY5Y cells. SH-SY5Y cells were differentiated using 10 µM all-trans retinoic acid for up to seven days. Neuronal differentiation was evaluated by morphological assessment and Western blot analysis of the neuronal markers MAP2 and Nestin. Retinoic acid treatment induced characteristic neuronal morphological changes, including neurite extension and reduced cellular clustering, consistent with successful differentiation. Endpoint PCR was performed to determine the presence of zDHHC transcripts, and quantitative real-time PCR was used to evaluate changes in gene expression throughout differentiation. Gene expression was normalized to GAPDH and β-actin. This study provides a comprehensive baseline characterization of zDHHC gene expression during RA-induced differentiation of SH-SY5Y cells and establishes a molecular framework for future investigations of zDHHC function and optimization of neuronal differentiation protocols.

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