Nanoscale architecture of synaptic vesicles and scaffolding complexes revealed by cryo-electron tomography

Author:

Held Richard G.12345ORCID,Liang Jiahao12345ORCID,Brunger Axel T.12345ORCID

Affiliation:

1. Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA 94305

2. Department of Neurology and Neurological Sciences, Stanford University, Stanford, CA 94305

3. Department of Structural Biology, Stanford University, Stanford, CA 94305

4. Department of Photon Science, Stanford University, Stanford, CA 94305

5. HHMI, Stanford University, Stanford, CA 94305

Abstract

The spatial distribution of proteins and their arrangement within the cellular ultrastructure regulates the opening of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors in response to glutamate release at the synapse. Fluorescence microscopy imaging revealed that the postsynaptic density (PSD) and scaffolding proteins in the presynaptic active zone (AZ) align across the synapse to form a trans-synaptic “nanocolumn,” but the relation to synaptic vesicle release sites is uncertain. Here, we employ focused-ion beam (FIB) milling and cryoelectron tomography to image synapses under near-native conditions. Improved image contrast, enabled by FIB milling, allows simultaneous visualization of supramolecular nanoclusters within the AZ and PSD and synaptic vesicles. Surprisingly, membrane-proximal synaptic vesicles, which fuse to release glutamate, are not preferentially aligned with AZ or PSD nanoclusters. These synaptic vesicles are linked to the membrane by peripheral protein densities, often consistent in size and shape with Munc13, as well as globular densities bridging the synaptic vesicle and plasma membrane, consistent with prefusion complexes of SNAREs, synaptotagmins, and complexin. Monte Carlo simulations of synaptic transmission events using biorealistic models guided by our tomograms predict that clustering AMPARs within PSD nanoclusters increases the variability of the postsynaptic response but not its average amplitude. Together, our data support a model in which synaptic strength is tuned at the level of single vesicles by the spatial relationship between scaffolding nanoclusters and single synaptic vesicle fusion sites.

Funder

Foundation for the National Institutes of Health

Howard Hughes Medical Institute

Publisher

Proceedings of the National Academy of Sciences

Cited by 2 articles. 订阅此论文施引文献 订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献

1. Taking a closer look at the synapse;Proceedings of the National Academy of Sciences;2024-08-05

2. Exploring the structural dynamics of the vesicle priming machinery;Biochemical Society Transactions;2024-07-31

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