Year of Award

2024

Document Type

Thesis

Degree Type

Master of Science (MS)

Other Degree Name/Area of Focus

Neuroscience

Department or School/College

Department of Biomedical and Pharmaceutical Sciences

Committee Chair

Jesse C. Hay

Commitee Members

Mark Grimes, Kasper Hansen, J. Steven Lodmell

Keywords

ER to Golgi transport, calcium signaling, ALG-2, peflin, fluorescent proteins, fluorescent imaging

Publisher

University of Montana

Subject Categories

Cell Biology | Molecular and Cellular Neuroscience

Abstract

The endoplasmic reticulum performs multiple functions in the eurkaryotic cell: biomolecule synthesis, protein folding, cargo sorting and secretion, and helping to maintain cellular calcium homeostasis. Though much has been discovered about these processes and the myriad proteins, enzymes, transcription factors, ions and other factors that play their own parts, much is still left to learn. To facilitate continued study of ER-to-Golgi transport, an array of genetically-encoded fluorescent tools are available to researchers. The featured review in Chapter 1 details the history of these tools and highlights some of the newest, which also incorporate information from the fields of synthetic chemistry and membrane dynamics. Tools that can be successfully used in neurons are specifically noted. Our lab utilizes many of these techniques in our own studies of ER-to-Golgi transport. In the paper in Chapter 2, we studied the penta-EF hand proteins ALG-2 and peflin as they interact with each other in the regulation of COPII-coated vesicle secretion, under different calcium signaling conditions. Our studies were conducted in 3 cell types: epithelial, fibroblast, and neuroendocrine, and our results showed that rate of secretion varied not only based on presence or absence of peflin, but also based on strength and duration of the calcium signal. In addition, the mobilization of cellular calcium through incubation with ATP produced surprising opposite effects on secretion and recruitment of COPII coat proteins in epithelial NRK cells versus undifferentiated neuroendocrine PC12 cells. These results confirm that COPII-associated ALG-2 is a dynamic regulator of ER-to-Golgi transport.

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© Copyright 2024 Danette M. Kowal Seiler