Cryo-electron tomographic investigation of native hippocampal glutamatergic synapses

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bioRxiv. 2024 Apr 1;. doi: 10.1101/2024.04.01.587595

Cryo-electron tomographic investigation of native hippocampal glutamatergic synapses Cryo-Electron Microscopy

Matsui Aya, Spangler Catherine, Elferich Johannes, Shiozaki Momoko, Jean Nikki, Zhao Xiaowei, Qin Maozhen, Zhong Haining, Yu Zhiheng, Gouaux Eric

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Abstract

Chemical synapses are the major sites of communication between neurons in the nervous system and mediate either excitatory or inhibitory signaling. At excitatory synapses, glutamate is the primary neurotransmitter and upon release from presynaptic vesicles, is detected by postsynaptic glutamate receptors, which include ionotropic AMPA and NMDA receptors. Here we have developed methods to identify glutamatergic synapses in brain tissue slices, label AMPA receptors with small gold nanoparticles (AuNPs), and prepare lamella for cryo-electron tomography studies. The targeted imaging of glutamatergic synapses in the lamella is facilitated by fluorescent pre- and postsynaptic signatures, and the subsequent tomograms allow for identification of key features of chemical synapses, including synaptic vesicles, the synaptic cleft and AuNP-labeled AMPA receptors. These methods pave the way for imaging natively derived brain regions at high resolution, using unstained, unfixed samples preserved under near-native conditions.

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Cryo-electron tomographic investigation of native hippocampal glutamatergic synapses

Abstract

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