A Versatile Synthetic Affinity Probe Reveals Inhibitory Synapse Ultrastructure and Brain Connectivity** - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue (Data Paper) Angewandte Chemie International Edition Année : 2022

A Versatile Synthetic Affinity Probe Reveals Inhibitory Synapse Ultrastructure and Brain Connectivity**

Vladimir Khayenko
Clemens Schulte
Sara Reis
  • Fonction : Auteur
Orly Avraham
  • Fonction : Auteur
Cataldo Schietroma
Rafael Worschech
  • Fonction : Auteur
Noah Nordblom
  • Fonction : Auteur
Sonja Kachler
  • Fonction : Auteur
Carmen Villmann
  • Fonction : Auteur
Katrin Heinze
  • Fonction : Auteur
Andreas Schlosser
  • Fonction : Auteur
Ora Schueler-Furman
Philip Tovote
Hans Maric

Résumé

Abstract Visualization of inhibitory synapses requires protocol tailoring for different sample types and imaging techniques, and usually relies on genetic manipulation or the use of antibodies that underperform in tissue immunofluorescence. Starting from an endogenous ligand of gephyrin, a universal marker of the inhibitory synapse, we developed a short peptidic binder and dimerized it, significantly increasing affinity and selectivity. We further tailored fluorophores to the binder, yielding “Sylite”—a probe with outstanding signal‐to‐background ratio that outperforms antibodies in tissue staining with rapid and efficient penetration, mitigation of staining artifacts, and simplified handling. In super‐resolution microscopy Sylite precisely localizes the inhibitory synapse and enables nanoscale measurements. Sylite profiles inhibitory inputs and synapse sizes of excitatory and inhibitory neurons in the midbrain and combined with complimentary tracing techniques reveals the synaptic connectivity.
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Origine : Publication financée par une institution
Licence : CC BY - Paternité

Dates et versions

hal-04252751 , version 1 (21-10-2023)

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Citer

Vladimir Khayenko, Clemens Schulte, Sara Reis, Orly Avraham, Cataldo Schietroma, et al.. A Versatile Synthetic Affinity Probe Reveals Inhibitory Synapse Ultrastructure and Brain Connectivity**. Angewandte Chemie International Edition, 2022, 61 (30), ⟨10.1002/anie.202202078⟩. ⟨hal-04252751⟩
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