학술논문

Genetic Probe for Visualizing Glutamatergic Synapses and Vesicles by 3D Electron Microscopy
Document Type
article
Source
ACS Chemical Neuroscience. 12(4)
Subject
Biochemistry and Cell Biology
Biological Sciences
Mental Health
Neurosciences
Animals
Glutamic Acid
Mice
Microscopy
Electron
Neurons
Synapses
Synaptic Vesicles
Vesicular Glutamate Transport Protein 1
Vesicular Glutamate Transport Protein 2
miniSOG
glutamatergic synapse
glutamate
vesicular glutamate transporter
neurotransmission
3D electron microscopy
deep learning
CDeep3M
genetic EM probe
ventral tegmental area
synaptic vesicles
Medicinal and Biomolecular Chemistry
Biochemistry and cell biology
Analytical chemistry
Medicinal and biomolecular chemistry
Language
Abstract
Communication between neurons relies on the release of diverse neurotransmitters, which represent a key-defining feature of a neuron's chemical and functional identity. Neurotransmitters are packaged into vesicles by specific vesicular transporters. However, tools for labeling and imaging synapses and synaptic vesicles based on their neurochemical identity remain limited. We developed a genetically encoded probe to identify glutamatergic synaptic vesicles at the levels of both light and electron microscopy (EM) by fusing the mini singlet oxygen generator (miniSOG) probe to an intralumenal loop of the vesicular glutamate transporter-2. We then used a 3D imaging method, serial block-face scanning EM, combined with a deep learning approach for automatic segmentation of labeled synaptic vesicles to assess the subcellular distribution of transporter-defined vesicles at nanometer scale. These tools represent a new resource for accessing the subcellular structure and molecular machinery of neurotransmission and for transmitter-defined tracing of neuronal connectivity.