A Versatile Synaptotagmin-1 Nanobody Provides Perturbation-Free Live Synaptic Imaging And Low Linkage-Error in Super-Resolution Microscopy.

calcium imaging intrabody nanobody sdAb super-resolution microscopy synaptic sensors

Journal

Small methods
ISSN: 2366-9608
Titre abrégé: Small Methods
Pays: Germany
ID NLM: 101724536

Informations de publication

Date de publication:
10 2023
Historique:
revised: 09 06 2023
received: 21 02 2023
medline: 23 10 2023
pubmed: 8 7 2023
entrez: 8 7 2023
Statut: ppublish

Résumé

Imaging of living synapses has relied for over two decades on the overexpression of synaptic proteins fused to fluorescent reporters. This strategy alters the stoichiometry of synaptic components and ultimately affects synapse physiology. To overcome these limitations, here a nanobody is presented that binds the calcium sensor synaptotagmin-1 (NbSyt1). This nanobody functions as an intrabody (iNbSyt1) in living neurons and is minimally invasive, leaving synaptic transmission almost unaffected, as suggested by the crystal structure of the NbSyt1 bound to Synaptotagmin-1 and by the physiological data. Its single-domain nature enables the generation of protein-based fluorescent reporters, as showcased here by measuring spatially localized presynaptic Ca

Identifiants

pubmed: 37421204
doi: 10.1002/smtd.202300218
doi:

Substances chimiques

Calcium SY7Q814VUP

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

e2300218

Subventions

Organisme : Deutsche Forschungsgemeinschaft
ID : SFB1286 (Z04)
Organisme : Deutsche Forschungsgemeinschaft
ID : FO 1342/1-3
Organisme : Schram Stiftung
ID : T0287/35359/2020
Organisme : MAX IV Laboratory for time on Beamline BioMAX under Proposal
ID : 20190233
Organisme : Swedish Research Council under contract
ID : 2018-07152
Organisme : Swedish Governmental Agency for Innovation Systems
ID : 2018-04969
Organisme : Swedish Governmental Agency for Innovation Systems
ID : 2019-02496
Organisme : Novo Nordisk Foundation
ID : NNF20OC0064789
Organisme : Swedish Research Council
ID : 2022-03681
Organisme : Swedish Cancer Society
ID : 20 1287 PjF

Informations de copyright

© 2023 The Authors. Small Methods published by Wiley-VCH GmbH.

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Auteurs

Karine Queiroz Zetune Villa Real (K)

Institute of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073, Göttingen, Germany.
Center for Biostructural Imaging of Neurodegeneration (BIN), University Medical Center Göttingen, 37075, Göttingen, Germany.

Nikolaos Mougios (N)

Institute of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073, Göttingen, Germany.
Center for Biostructural Imaging of Neurodegeneration (BIN), University Medical Center Göttingen, 37075, Göttingen, Germany.

Ronja Rehm (R)

Institute of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073, Göttingen, Germany.

Shama Sograte-Idrissi (S)

Institute of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073, Göttingen, Germany.
Center for Biostructural Imaging of Neurodegeneration (BIN), University Medical Center Göttingen, 37075, Göttingen, Germany.

László Albert (L)

Institute of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073, Göttingen, Germany.
Center for Biostructural Imaging of Neurodegeneration (BIN), University Medical Center Göttingen, 37075, Göttingen, Germany.

Amir Mohammad Rahimi (AM)

Institute of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073, Göttingen, Germany.
Center for Biostructural Imaging of Neurodegeneration (BIN), University Medical Center Göttingen, 37075, Göttingen, Germany.

Manuel Maidorn (M)

Institute of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073, Göttingen, Germany.
Center for Biostructural Imaging of Neurodegeneration (BIN), University Medical Center Göttingen, 37075, Göttingen, Germany.

Jannik Hentze (J)

Institute of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073, Göttingen, Germany.
Center for Biostructural Imaging of Neurodegeneration (BIN), University Medical Center Göttingen, 37075, Göttingen, Germany.

Markel Martínez-Carranza (M)

Department of Biochemistry and Biophysics, Stockholm University, Stockholm, Stockholm, SE-10691, Sweden.

Hassan Hosseini (H)

Research Group Neuromodulatory Networks, Leibniz Institute for Neurobiology, 39118, Magdeburg, Germany.

Kim-Ann Saal (KA)

Institute of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073, Göttingen, Germany.

Nazar Oleksiievets (N)

III. Institute of Physics - Biophysics, Georg August University, 37077, Göttingen, Germany.

Matthias Prigge (M)

Research Group Neuromodulatory Networks, Leibniz Institute for Neurobiology, 39118, Magdeburg, Germany.
Center for Behavioral Brain Sciences, 39118, Magdeburg, Germany.

Roman Tsukanov (R)

III. Institute of Physics - Biophysics, Georg August University, 37077, Göttingen, Germany.

Pål Stenmark (P)

Department of Biochemistry and Biophysics, Stockholm University, Stockholm, Stockholm, SE-10691, Sweden.

Eugenio F Fornasiero (EF)

Institute of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073, Göttingen, Germany.

Felipe Opazo (F)

Institute of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073, Göttingen, Germany.
Center for Biostructural Imaging of Neurodegeneration (BIN), University Medical Center Göttingen, 37075, Göttingen, Germany.
NanoTag Biotechnologies GmbH, 37079, Göttingen, Germany.

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