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

  • Author / Creator
    Nilsson, Benjamin G
  • 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.

  • Subjects / Keywords
  • Graduation date
  • Type of Item
  • Degree
    Master of Science
  • DOI
  • License
    This thesis is made available by the University of Alberta Libraries with permission of the copyright owner solely for non-commercial purposes. This thesis, or any portion thereof, may not otherwise be copied or reproduced without the written consent of the copyright owner, except to the extent permitted by Canadian copyright law.
  • Language
  • Institution
    University of Alberta
  • Degree level
  • Department
    • Department of Pharmacology
  • Supervisor / co-supervisor and their department(s)
    • Dunn, Susan (Pharmacology)
    • Holt, Andy (Pharmacology)
    • Kozuska, Janna (Pharmacology)
  • Examining committee members and their departments
    • Tse, Fred (Pharmacology)
    • Casey, Joe (Phyiology)
    • Baker, Glen (Psychiatry)