Date of Award

1-1-2022

Language

English

Document Type

Master's Thesis

Degree Name

Master of Science (MS)

College/School/Department

Department of Biological Sciences

Content Description

1 online resource (v, 28 pages)

Dissertation/Thesis Chair

Annalisa Scimemi

Committee Members

Sarah McCallum

Keywords

Neurotransmitters, Proteins, Fluorescent polymers

Subject Categories

Biology

Abstract

Neurons in the brain communicate via electrical and chemical synapses, whereneurotransmitters are released. Traditional methods to detect synaptic transmission rely on single-cell patch-clamp recordings from target neurons, but these can only provide an indirect readout of the lifetime of neurotransmitters in the extracellular space. More recently, this task has been aided by developing fluorescent biosensors with high sensitivity and large dynamic ranges. Fluorescent biosensors are typically composed of a neurotransmitter binding protein tagged with fluorescent reporter molecules. Fluorescence biosensors come in different forms, including FRET-based biosensors. These can be built using different intracellular signaling molecules and neurotransmitter binding domains. Here, I describe the operational concepts of current intensiometric and ratiometric biosensors and provide an understanding of the neurotransmitter-binding proteins from bacteria and eukaryotes, including periplasmic-binding proteins and G-protein coupled receptors which serve as scaffolds. Next, I describe the blueprint for developing neurotransmitter biosensors and their variants. Here, I highlight the general application of these biosensors to shed light on the functionality of neurotransmitters. Furthermore, some improvements need to be made to overcome technical problems, and there are promising and exciting significant advances for the future of fluorescent biosensors, such as in vivo multicolor imaging.

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