Region-Specific Phosphorylation Determines Neuroligin-3 Localization to Excitatory Versus Inhibitory Synapses

Overview

Journal Biol Psychiatry

Publisher Elsevier

Specialty Psychiatry

Date 2023 Dec 28

PMID 38154503

Authors

Bekir Altas

Liam P Tuffy

Annarita Patrizi

Kalina Dimova

Tolga Soykan

Cheryl Brandenburg

Andrea J Romanowski

Julia R Whitten

Colin D Robertson

Saovleak N Khim

Garrett W Crutcher

Mateusz C Ambrozkiewicz

Oleksandr Yagensky

Dilja Krueger-Burg

Matthieu Hammer

He-Hsuan Hsiao

Pawel R Laskowski

Lydia Dyck

Adam C Puche

Marco Sassoe-Pognetto

John J E Chua

Henning Urlaub

Olaf Jahn

Nils Brose

Alexandros Poulopoulos

Affiliations

Soon will be listed here.

Abstract

Background: Neuroligin-3 is a postsynaptic adhesion molecule involved in synapse development and function. It is implicated in rare, monogenic forms of autism, and its shedding is critical to the tumor microenvironment of gliomas. While other members of the neuroligin family exhibit synapse-type specificity in localization and function through distinct interactions with postsynaptic scaffold proteins, the specificity of neuroligin-3 synaptic localization remains largely unknown.

Methods: We investigated the synaptic localization of neuroligin-3 across regions in mouse and human brain samples after validating antibody specificity in knockout animals. We raised a phospho-specific neuroligin antibody and used phosphoproteomics, cell-based assays, and in utero CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/Cas9) knockout and gene replacement to identify mechanisms that regulate neuroligin-3 localization to distinct synapse types.

Results: Neuroligin-3 exhibits region-dependent synapse specificity, largely localizing to excitatory synapses in cortical regions and inhibitory synapses in subcortical regions of the brain in both mice and humans. We identified specific phosphorylation of cortical neuroligin-3 at a key binding site for recruitment to inhibitory synapses, while subcortical neuroligin-3 remained unphosphorylated. In vitro, phosphomimetic mutation of that site disrupted neuroligin-3 association with the inhibitory postsynaptic scaffolding protein gephyrin. In vivo, phosphomimetic mutants of neuroligin-3 localized to excitatory postsynapses, while phospho-null mutants localized to inhibitory postsynapses.

Conclusions: These data reveal an unexpected region-specific pattern of neuroligin-3 synapse specificity, as well as a phosphorylation-dependent mechanism that regulates its recruitment to either excitatory or inhibitory synapses. These findings add to our understanding of how neuroligin-3 is involved in conditions that may affect the balance of excitation and inhibition.

Citing Articles

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PMID: 38298376 PMC: 10827848. DOI: 10.3389/fncel.2023.1332179.

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