Genetic Probe for Visualizing Glutamatergic Synapses and Vesicles by 3D Electron Microscopy.
Thomas Steinkellner1,2, Matthew Madany1,3, Matthias G Haberl1,3,4
1Department of Neurosciences, University of California, San Diego, La Jolla, California 92093, United States.
ACS Chemical Neuroscience
|February 1, 2021
Summary
Researchers developed a new genetic probe to visualize glutamatergic synaptic vesicles using microscopy. This tool aids in studying neurotransmission and neuronal connectivity at the nanoscale.
Area of Science:
- Neuroscience
- Cell Biology
- Molecular Biology
Background:
- Neuronal communication depends on neurotransmitters, defining neuron identity.
- Vesicular transporters package neurotransmitters, but tools for neurochemical identification are limited.
Purpose of the Study:
- To develop a genetically encoded probe for labeling glutamatergic synaptic vesicles.
- To enable visualization at both light and electron microscopy (EM) levels.
- To facilitate nanoscale analysis of vesicle distribution and neuronal connectivity.
Main Methods:
- Genetically encoded probe: mini singlet oxygen generator (miniSOG) fused to vesicular glutamate transporter-2.
- 3D imaging: serial block-face scanning EM.
- Deep learning for automatic segmentation of labeled synaptic vesicles.
Main Results:
- Successful labeling and imaging of glutamatergic synaptic vesicles.
- Nanoscale assessment of subcellular distribution of transporter-defined vesicles.
- Demonstration of a new resource for studying neurotransmission machinery.
Conclusions:
- The developed probe and imaging methods provide novel tools for neuroscience research.
- Enables detailed analysis of synaptic vesicle distribution and neuronal connectivity.
- Advances understanding of the molecular machinery underlying neurotransmission.
