Publication Date

2011-05-02

Availability

Embargoed

Embargo Period

2013-05-01

Degree Name

Master of Science (MS)

Department

Chemistry (Arts and Sciences)

Date of Defense

2011-04-12

First Committee Member

James N. Wilson

Second Committee Member

Angel E. Kaifer

Third Committee Member

Norito Takenaka

Abstract

Fluorescent analogs of biomolecules have been known as useful probes to study the structure, conformations and dynamics of cellular processes. These probes are more ideal than fluorescent labeled probes, as fluorescent analog probes retain the shape, size, conformation, and recognition element of the natural substrate, while giving useful intracellular information about detection and dynamics of biomolecules. The monoamine neurotransmitters control the central and periphery nervous systems. Serotonin (5-HT), in particular, is a versatile chemical messenger responsible for a multitude of biological processes, such as regulation of emotion, vasoconstriction, and bone metabolism. The study of serotonergic complex pathways is vital and essential in drug discovery for the diseases that result from the depletion and deregulation of serotonin in synapse. The extracellular concentration of serotonin is controlled by several transporters, most preferably the serotonin transporter (SERT). Selective serotonin reuptake inhibitors (SSRIs), along with dual- and triple-acting inhibitors, affect SERT and hence 5-HT in depression and related diseases. In this present investigation, firstly, a set of fluorescent analogs of neurotransmitter probes based on ethylamino-functionalized substrates were successfully designed and these fluorescent probes were synthesized by convenient synthetic methods. Secondly, optical properties of these fluorescent probes were investigated in organic medium, in order to test their suitability for screening and imaging the biological cells. Finally, their uptake was examined in the murine osteocytic cell line, MLO-Y4, platelets of blood sample and HEK-293 cells expressing the dopamine transporter (DAT), norepinephrine transporter (NET) or SERT. The fluorescent probes targeting bone-derived cell line expressing 5-HTT provide useful information in understanding the dynamics of 5-HT regulation with respect to SSRI treatment. A novel fluorescent analog of 5-HT probe was developed that may be utilized to study 5-HTT function in the context of 5-HT uptake or regulation in cell culture, tissue explants, or even in vivo.

Keywords

Fluorescent; Cellular; Neurotransmitters; Inhibition; Monoamine; Uptake.

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