Genetic Probe for Visualizing Glutamatergic Synapses and Vesicles by 3D Electron Microscopy.

Thomas Steinkellner, Matthew Madany, Matthias G Haberl, Vivien Zell, Carolina Li, Junru Hu, Mason Mackey, Ranjan Ramachandra, Stephen Adams, Mark H Ellisman, Thomas S Hnasko, Daniela Boassa

ACS chemical neuroscience 2021 Feb 17

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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.

Citation

Thomas Steinkellner, Matthew Madany, Matthias G Haberl, Vivien Zell, Carolina Li, Junru Hu, Mason Mackey, Ranjan Ramachandra, Stephen Adams, Mark H Ellisman, Thomas S Hnasko, Daniela Boassa. Genetic Probe for Visualizing Glutamatergic Synapses and Vesicles by 3D Electron Microscopy. ACS chemical neuroscience. 2021 Feb 17;12(4):626-639