Imaging data for inhibitory synaptic vesicles
Abstract/Contents
- Abstract
- Synaptic vesicles specific to inhibitory GABA-releasing neurons are critical for regulating neuronal excitability. To study the specific molecular composition, architecture, and function of inhibitory synaptic vesicles, we have developed a new method to isolate and purify GABA synaptic vesicles from mouse brains. GABA synaptic vesicles were immunoisolated from mouse brain tissue using an engineered fragment antigen-binding region (Fab) against the vesicular GABA transporter (vGAT) and purified. The GABA synaptic vesicles were specifically enriched for vGAT and largely depleted of contaminants from other synaptic vesicle types, such as vesicular glutamate transporter (vGLUT1), and other cellular organelles. This degree of purity was achieved despite the relatively low abundance of vGAT vesicles compared to the total synaptic vesicle pool in mammalian brains. Cryo-electron microscopy images of these isolated GABA synaptic vesicles revealed intact morphology with circular shape and protruding proteinaceous densities. The GABA synaptic vesicles are functional, as assessed by a hybrid (ex vivo/in vitro) vesicle fusion assay, and they undergo synchronized fusion with synthetic plasma membrane mimic vesicles in response to Ca2+-triggering, but, as a negative control, not to Mg2+-triggering. Our immunoisolation method could also be applied to other types of vesicles. This repository contains the imaging data for the single vesicle association and fusion experiments, and all micrographs for the negative stain and cryo-EM experiments.
Description
| Type of resource | Dataset, still image |
|---|---|
| Date created | [ca. May 2022 - August 2023] |
| Date modified | March 10, 2024; March 15, 2024 |
| Publication date | February 23, 2024; March 10, 2024 |
Creators/Contributors
| Author | Gopal, Nisha |
https://orcid.org/0000-0003-1980-2292
(unverified)
|
|---|---|---|
| Author | Brunger, Axel |
https://orcid.org/0000-0001-5121-2036
(unverified)
|
Subjects
| Subject | synaptic vesicle isolation |
|---|---|
| Subject | fragment antigen-binding region (Fab) |
| Subject | vGAT |
| Subject | vesicular GABA transporter |
| Subject | fusion assay |
| Genre | Data |
| Genre | Image |
| Genre | Data sets |
| Genre | Dataset |
Bibliographic information
| Related item | |
|---|---|
| DOI | https://doi.org/10.25740/ht023wd6382 |
| Location | https://purl.stanford.edu/ht023wd6382 |
Access conditions
- Use and reproduction
- User agrees that, where applicable, content will not be used to identify or to otherwise infringe the privacy or confidentiality rights of individuals. Content distributed via the Stanford Digital Repository may be subject to additional license and use restrictions applied by the depositor.
- License
- This work is licensed under a Creative Commons Attribution 4.0 International license (CC BY).
Preferred citation
- Preferred citation
- Gopal, N. and Brunger, A. (2023). Imaging data for inhibitory synaptic vesicles. Stanford Digital Repository. Available at https://purl.stanford.edu/ht023wd6382. https://doi.org/10.25740/ht023wd6382.
Collection
Stanford Research Data
View other items in this collection in SearchWorksContact information
Also listed in
Loading usage metrics...