3D Super-Resolution Imaging of PSD95 Reveals an Abundance of Diffuse Protein Supercomplexes in the Mouse Brain

ACS Chemical Neuroscience, Journal Year: 2024, Volume and Issue: 16(1), P. 40 - 51

Published: Dec. 20, 2024

PSD95 is an abundant scaffolding protein that assembles multiprotein complexes controlling synaptic physiology and behavior. Confocal microscopy has previously shown enriched in the postsynaptic terminals of excitatory synapses two-dimensional (2D) super-resolution further revealed it forms nanoclusters. In this study, we utilized three-dimensional (3D) to examine nanoarchitecture mouse brain, characterizing spatial arrangement over 8 million molecules. While were able identify molecular arrangements have been reported, imaging 3D allowed us classify these with higher accuracy. Furthermore, enabled quantification levels, revealing abundance molecules existed outside as a diffuse population supercomplexes, containing multiple copies PSD95. Further analysis supercomplexes two units identified populations: one had separated by 39 ± 2 nm, second separation 94 27 nm. The finding there exists synapse suggests assemble then integrate into form supramolecular nanocluster architecture.

Language: Английский

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Expression and structural analysis of human neuroligin 2 and neuroligin 3 implicated in autism spectrum disorders

Frontiers in Endocrinology, Journal Year: 2022, Volume and Issue: 13

Published: Nov. 21, 2022

The development of autism spectrum disorders (ASDs) involves both environmental factors such as maternal diabetes and genetic neuroligins (NLGNs). NLGN2 NLGN3 are two members NLGNs with distinct distributions functions in synapse plasticity. relationship between NLGNs, the working mechanisms different currently remain unclear. Here, we first analyzed expression levels a streptozotocin-induced ASD mouse model brain regions to reveal their differences similarities. Then, cryogenic electron microscopy (cryo-EM) structures human were determined. overall similar homologs previous reports. However, structural comparisons revealed relative rotations protomers homodimers NLGN3. Taken together previously reported NLGN2–MDGA1 complex, speculate that assembly adopted by may affect interactions MDGAs. Our results provide insights into potential implicated ASD.

Language: Английский

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Trans-synaptic molecular context of NMDA receptor nanodomains

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Dec. 23, 2023

Tight coordination of the spatial relationships between protein complexes is required for cellular function. In neuronal synapses, many proteins responsible neurotransmission organize into subsynaptic nanoclusters whose trans-cellular alignment modulates synaptic signal propagation. However, these and NMDA receptors (NMDARs), which are learning memory, remain undefined. Here, we mapped relationship key NMDAR subunits to reference in active zone postsynaptic density using multiplexed super-resolution DNA-PAINT microscopy. GluN2A GluN2B formed with diverse configurations that, surprisingly, were not localized near presynaptic vesicle release sites marked by Munc13-1. a subset was configured maintain activation: internally denser, aligned abundant PSD-95, associated closely specific nanodomains. This work reveals new principle regulating signaling suggests that functional architecture depends on assembly multiprotein nanodomains interior construction conditional relationships.

Language: Английский

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CASK and FARP localize two classes of post-synaptic ACh receptors thereby promoting cholinergic transmission

PLoS Genetics, Journal Year: 2022, Volume and Issue: 18(10), P. e1010211 - e1010211

Published: Oct. 24, 2022

Changes in neurotransmitter receptor abundance at post-synaptic elements play a pivotal role regulating synaptic strength. For this reason, there is significant interest identifying and characterizing the scaffolds required for localization different synapses. Here we analyze of two C . elegans scaffolding proteins (LIN-2/CASK FRM-3/FARP) cholinergic neuromuscular junctions. Constitutive knockouts or muscle specific inactivation lin-2 frm-3 dramatically reduced spontaneous evoked currents. These defects resulted from decreased classes ionotropic acetylcholine receptors (ACR-16/CHRNA7 levamisole-activated AChRs). LIN-2’s AChR function mediated by its SH3 PDZ domains, which interact with AChRs FRM-3/FARP, respectively. Thus, our findings show that LIN-2/FRM-3 complexes promote transmission recruiting to elements.

Language: Английский

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The Essential Role of Latrophilin-1 Adhesion GPCR Nanoclusters in Inhibitory Synapses

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Oct. 8, 2023

ABSTRACT Latrophilin-1 (Lphn1, a.k.a. CIRL1 and CL1; gene symbol Adgrl1 ) is an Adhesion GPCR that has been implicated in excitatory synaptic transmission as a candidate receptor for α-latrotoxin. Here we analyzed conditional knockin/knockout mice Lphn1 contain extracellular myc-epitope tag. Surprisingly, found localized cultured neurons to nanoclusters are present both inhibitory synapses. Conditional deletion of failed elicit detectable impairment synapses but produced decrease synapse numbers was most pronounced close the neuronal soma. No changes axonal or dendritic outgrowth branching were observed. Our data indicate among few postsynaptic adhesion molecules by itself not essential contributes connections.

Language: Английский

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