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

Neuroligin autism excitatory synapse inhibitory synapse phosphorylation scaffolding protein

Journal

Biological psychiatry
ISSN: 1873-2402
Titre abrégé: Biol Psychiatry
Pays: United States
ID NLM: 0213264

Informations de publication

Date de publication:
26 Dec 2023
Historique:
received: 29 09 2023
revised: 19 12 2023
accepted: 19 12 2023
medline: 29 12 2023
pubmed: 29 12 2023
entrez: 28 12 2023
Statut: aheadofprint

Résumé

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. 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 knockout and gene replacement to identify mechanisms that regulate Neuroligin-3 localization to distinct synapse types. 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. 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.

Sections du résumé

BACKGROUND 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 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 knockout and gene replacement to identify mechanisms that regulate Neuroligin-3 localization to distinct synapse types.
RESULTS 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 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.

Identifiants

DOI: 10.1016/j.biopsych.2023.12.020 PMID: 38154503
pubmed: 38154503
pii: S0006-3223(23)01799-7
doi: 10.1016/j.biopsych.2023.12.020
pii:
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Informations de copyright

Copyright © 2023. Published by Elsevier Inc.

Auteurs

Bekir Altas (B)

University of Maryland School of Medicine, Department of Pharmacology, Baltimore, MD 21201, USA.

Liam P Tuffy (LP)

Max Planck Institute for Multidisciplinary Sciences, Department of Molecular Neurobiology, 37075 Göttingen, Germany.

Annarita Patrizi (A)

University of Turin, Department of Neuroscience "Rita Levi Montalcini", I-10126 Turin, Italy.

Kalina Dimova (K)

Max Planck Institute for Multidisciplinary Sciences, Department of Molecular Neurobiology, 37075 Göttingen, Germany; Max Planck Institute for Multidisciplinary Sciences, Department of Molecular Neurobiology, Neuroproteomics Group, 37075 Göttingen, Germany.

Tolga Soykan (T)

Max Planck Institute for Multidisciplinary Sciences, Department of Molecular Neurobiology, 37075 Göttingen, Germany.

Cheryl Brandenburg (C)

University of Maryland School of Medicine, Department of Pharmacology, Baltimore, MD 21201, USA.

Andrea J Romanowski (AJ)

University of Maryland School of Medicine, Department of Pharmacology, Baltimore, MD 21201, USA.

Julia R Whitten (JR)

University of Maryland School of Medicine, Department of Pharmacology, Baltimore, MD 21201, USA.

Colin D Robertson (CD)

University of Maryland School of Medicine, Department of Pharmacology, Baltimore, MD 21201, USA.

Saovleak N Khim (SN)

University of Maryland School of Medicine, Department of Pharmacology, Baltimore, MD 21201, USA.

Garrett W Crutcher (GW)

University of Maryland School of Medicine, Department of Pharmacology, Baltimore, MD 21201, USA.

Mateusz C Ambrozkiewicz (MC)

Max Planck Institute for Multidisciplinary Sciences, Department of Molecular Neurobiology, 37075 Göttingen, Germany.

Oleksandr Yagensky (O)

Max Planck Institute for Multidisciplinary Sciences, Laboratory of Neurobiology, Research Group Protein Trafficking in Synaptic Development and Function, 37077 Göttingen, Germany.

Dilja Krueger-Burg (D)

Max Planck Institute for Multidisciplinary Sciences, Department of Molecular Neurobiology, 37075 Göttingen, Germany.

Matthieu Hammer (M)

Max Planck Institute for Multidisciplinary Sciences, Department of Molecular Neurobiology, 37075 Göttingen, Germany.

He-Hsuan Hsiao (HH)

Max Planck Institute for Multidisciplinary Sciences, Bioanalytical Mass Spectrometry, 37077 Göttingen, Germany.

Pawel R Laskowski (PR)

Max Planck Institute for Multidisciplinary Sciences, Department of Molecular Neurobiology, 37075 Göttingen, Germany.

Lydia Dyck (L)

Max Planck Institute for Multidisciplinary Sciences, Department of Molecular Neurobiology, 37075 Göttingen, Germany.

Adam C Puche (AC)

University of Maryland School of Medicine, Department of Neurobiology, Baltimore, MD 21201, USA.

Marco Sassoè-Pognetto (M)

University of Turin, Department of Neuroscience "Rita Levi Montalcini", I-10126 Turin, Italy.

John J E Chua (JJE)

Max Planck Institute for Multidisciplinary Sciences, Laboratory of Neurobiology, Research Group Protein Trafficking in Synaptic Development and Function, 37077 Göttingen, Germany.

Henning Urlaub (H)

Max Planck Institute for Multidisciplinary Sciences, Bioanalytical Mass Spectrometry, 37077 Göttingen, Germany; University Medical Center Göttingen, Institute of Clinical Chemistry, Bioanalytics Group, 37075 Göttingen, Germany.

Olaf Jahn (O)

Max Planck Institute for Multidisciplinary Sciences, Department of Molecular Neurobiology, Neuroproteomics Group, 37075 Göttingen, Germany; University Medical Center Göttingen, Department of Psychiatry and Psychotherapy, Translational Neuroproteomics Group, 37075 Göttingen, Germany.

Nils Brose (N)

Max Planck Institute for Multidisciplinary Sciences, Department of Molecular Neurobiology, 37075 Göttingen, Germany.

Alexandros Poulopoulos (A)

University of Maryland School of Medicine, Department of Pharmacology, Baltimore, MD 21201, USA; Max Planck Institute for Multidisciplinary Sciences, Department of Molecular Neurobiology, 37075 Göttingen, Germany. Electronic address: apoulopoulos@som.umaryland.edu.

Classifications MeSH