Dissertation
STUDIES OF SYNTHETIC PARTICLES AND NERVE ENDINGS ON MASS TRANSPORT AND KINETICS AND INHIBITION OF THE DEGLYCOSYLATED DOPAMINE TRANSPORTER
Doctor of Philosophy (PhD), Washington State University
01/2012
Handle:
https://hdl.handle.net/2376/4085
Abstract
Unstirred layer (USL) is present at the interface of solids with solutions, and particles pass through these layers by diffusion. Rotating disk electrode voltammetry (RDEV) has been used in different biological applications. However, it is unknown whether brain tissue preparation in solution affects the mass transport at the electrode, and whether the USL in real biological samples affects the kinetic measurements. In Chapter Two, liposomes, silica, and SephadexTM were used to model the tissue preparation particles and examined the effect on mass transport at the RDE. Both diffusion and hydrodynamic boundary layers formed between the solution and the electrode surface. In the presence of inert particles, the sensitivity, limiting current, apparent diffusion coefficient, boundary layers thicknesses, and the mixing time decreased with no change on the detection limit (6 ± 2 nM). In addition, the thicknesses of USL's within the neuronal homogenates do not appear to affect the apparent kinetics of biological mass transport. Results also show that RDEV kinetically resolves transmembrane transport with a timing of approximately 30 ms.
It is known that the glycosylation on the dopamine transporter (DAT) is important for cocaine binding and the function of the DAT. However, the connection between carbohydrate moieties and the function of the DAT is unknown. In Chapter Three, effects of carbohydrate component modification on DA transport velocity were examined by using different glycosidases. DA transport activities decreased after different glycosidase treatments in rats' striata. Then, α-mannosidase was used for the rest of the studies due to the rapid access to its substrate. After α-mannosidase treatment, Vmax of the DA transport decreased while the Km was unchanged. Moreover, removal of α-mannose abolished the effect of cocaine on the kinetic state of the DAT. Finally, cocaine, bupropion, mazindol, and β-phenylethylamine but not methamphetamine and tyramine block the effects of α-mannosidase on the DAT. Results lead to a conclusion that a derivative of mazindol may be a possible antagonist for cocaine.
Methamphetamine is a very addictive drug that will damage the brain. Chapter Four provides a summary on how methamphetamine affects different neurotransmitter contents and its transporter kinetics in different brain regions.
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Details
- Title
- STUDIES OF SYNTHETIC PARTICLES AND NERVE ENDINGS ON MASS TRANSPORT AND KINETICS AND INHIBITION OF THE DEGLYCOSYLATED DOPAMINE TRANSPORTER
- Creators
- Veronica Chiu
- Contributors
- James O Schenk (Advisor)Herbert H Hill (Committee Member)Chulhee Kang (Committee Member)Barbara A Sorg (Committee Member)
- Awarding Institution
- Washington State University
- Academic Unit
- Chemistry, Department of
- Theses and Dissertations
- Doctor of Philosophy (PhD), Washington State University
- Number of pages
- 149
- Identifiers
- 99900581656501842
- Language
- English
- Resource Type
- Dissertation