A structural and functional investigation of inhibitory synaptic transmission
Abstract/Contents
- Abstract
- Gamma-aminobutyric acid (GABA) is the primary inhibitory neurotransmitter of the mammalian central nervous system. Neurons release GABA to shunt or prevent action potentials from propagating in target cells, and this process is critical to providing a dynamic regulation of overall neural circuitry. Chapter 1: I briefly introduce neurotransmission and focus on inhibitory neurotransmission mediated by GABA signaling. Chapter 2: I describe the development of an anti-vGAT recombinant Fab domain used to purify intact inhibitory GABA SVs from mouse brains and the subsequent structural characterization of these vesicles using electron microscopy. I also describe the use of these vesicles in a functional assay that directly probes the membrane association and membrane fusion of GABA synaptic vesicles. The results in this chapter have been published in the journal Current Research in Physiology in their special edition issue on neurotransmitter transporters. Chapter 3: Following the success of the vGAT Fab domain in purifying GABASVs, I describe the development of a molecular toolbox for the isolation and study of other types of vesicular neurotransmitter transporters and SVs using the recombinant Fab domain approach. The results shared in this chapter are preliminary, and the work is ongoing. Chapter 4: I use the recombinant anti-vGAT Fab domain to isolate detergent-solubilized vGAT from rat brains for structural characterization via CryoEM. The results shared in this chapter are preliminary, and the work is ongoing. Chapter 5: I describe an orthogonal structural characterization of purified GABA synaptic vesicles using high-resolution CryoEM. More specifically, the structure of the native vacuolar ATPase was investigated in the context of GABA SV membranes to determine its direct binding partners. The results shared in this chapter are preliminary, and the work is ongoing.
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource. |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2024; ©2024 |
| Publication date | 2024; 2024 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Gopal, Nisha |
|---|---|
| Degree supervisor | Brünger, Axel T |
| Thesis advisor | Brünger, Axel T |
| Thesis advisor | Feng, Liang, 1976- |
| Thesis advisor | Reimer, Richard J |
| Thesis advisor | SShen, Kang, 1972- |
| Degree committee member | Feng, Liang, 1976- |
| Degree committee member | Reimer, Richard J |
| Degree committee member | SShen, Kang, 1972- |
| Associated with | Stanford University, School of Medicine |
| Associated with | Stanford University, Department of Structural Biology |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Nisha Gopal. |
|---|---|
| Note | Submitted to the Department of Structural Biology. |
| Thesis | Thesis Ph.D. Stanford University 2024. |
| Location | https://purl.stanford.edu/wg270qf2497 |
Access conditions
- Copyright
- © 2024 by Nisha Gopal
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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