Year of Publication

2020

Degree Name

Doctor of Philosophy (PhD)

Document Type

Doctoral Dissertation

College

Arts and Sciences

Department

Chemistry

First Advisor

Dr. Christopher I. Richards

Abstract

The consumption of tobacco products leads to several health risks, including respiratory diseases, cardiovascular diseases, and many different cancers. Nicotine, the primary addictive compound in tobacco, acts on the cholinergic system through nicotinic acetylcholine receptors (nAChRs), which is believed to be an essential component of addiction. However, the lack of a complete understanding of the neurobiological mechanisms of nicotine abuse is one of the primary barriers to finding potent therapeutics for smoking cessation. Single-molecule imaging is an ideal tool to study molecular mechanisms of protein and physiological changes both in vitro and in vivo. In this dissertation, our research efforts focused on the discovery of novel single-molecule imaging tools that aid in understanding the physiological changes related to nicotine addiction.

Specifically, we studied 1) membrane receptor assembly. We developed an innovative ex vivo approach that enables brain region specific single-molecule imaging to monitor the distribution of α4β2 nAChR assembly during nicotine exposure and withdrawal. This work reveals the selectivity of nicotine-induced upregulation in different brain regions in live animals. 2) characterization of nanovesicles derived from cells. We developed a high throughput fluorescence correlation spectroscopy (ht-FCS) approach that enables the rapid characterization of vesicle surface proteins. Characterized vesicles could either be explored as therapeutic delivery vehicles or be used to study membrane receptors. 3) blood flow properties. We demonstrated the application of multiphoton in vivo fluorescence correlation spectroscopy (FCS) for the measurement of cerebral blood flow with high spatial and temporal resolution. The cerebrovascular dysfunction is potentially linked to nicotine use disorder, blood-brain barrier (BBB) disruption, and gliovascular coupling.

Digital Object Identifier (DOI)

https://doi.org/10.13023/etd.2020.307

Available for download on Thursday, July 22, 2021

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