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Permanent link (DOI): https://doi.org/10.7939/R37659N64

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GABAA Receptors and Tonic Inhibition: Towards an Improved Understanding of Agonist Binding and in vivo Expression of the Extrasynaptic a4b3d Subtype Open Access

Descriptions

Other title
Subject/Keyword
GABA receptor
fluorescence resonance energy transfer
GABA
extrasynaptic
agonist binding
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Nilsson, Benjamin G
Supervisor and department
Dunn, Susan (Pharmacology)
Kozuska, Janna (Pharmacology)
Holt, Andy (Pharmacology)
Examining committee member and department
Baker, Glen (Psychiatry)
Casey, Joe (Phyiology)
Tse, Fred (Pharmacology)
Department
Department of Pharmacology
Specialization

Date accepted
2013-06-21T11:41:13Z
Graduation date
2013-11
Degree
Master of Science
Degree level
Master's
Abstract
The GABAA receptor is the major inhibitory neurotransmitter receptor in the central nervous system. Receptors containing the delta subunit generate tonic inhibition due to their extrasynaptic expression, high affinity for gamma-aminobutyric acid (GABA), and slow densensitization kinetics. The present work had two goals: first, compare structural elements involved in agonist binding in the a1b2g2 and a4b3d receptors, which are model synaptic and extrasynaptic receptor subtypes, respectively; second, develop an immunoassay using two-step fluorescence resonance energy transfer to detect the incorporation of three subunits into one receptor complex. The structural studies showed that the loop D region participates in agonist activity at both receptor subtypes, and that the agonist 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol (THIP) may function through a distinct subsite from that of GABA. Inadequate expression of the subunit constructs limited progress on the immunoassay, requiring more work to optimize the expression system before proceeding to proof-of-principle studies using two-step FRET.
Language
English
DOI
doi:10.7939/R37659N64
Rights
Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
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