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<p>Synaptic neurotransmission is a critical hallmark of brain activity and one of the first processes affected in neural diseases. Monitoring this process, particularly synaptic vesicle recycling, in living cells has been instrumental in revealing the mechanisms responsible for neurotransmitter release. However, currently available reporters suffer from limitations, such as large probe sizes or limited compatibility for human neurons, hampering the quantitative analysis of synaptic pathophysiology. Here, we describe the NbLumSyt1 toolkit, a panel of nanobody-based affinity probes that target the luminal domain of the synaptic vesicle protein Synaptotagmin 1 (Syt1). These new tools enable quantitative, noninvasive imaging and functional interrogation of Syt1 exo-endocytosis and trafficking in human neurons, with unprecedented precision, versatility and cost efficiency, in technologies ranging from fixed- and live-cell super-resolution imaging to electron microscopy and mass spectrometry. Overall, NbLumSyt1 nanobinders provide a valuable platform for studying synaptic physiology and pathophysiology, benefiting fundamental neuroscience and translational efforts to study and develop treatments for brain-related disorders.</p> Graphical Abstract <p></p>

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A versatile nanobody platform for live and super-resolution imaging of synaptic vesicle dynamics and plasticity in rodent and human neurons

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年5月14日

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  • Rashi Goel,
  • Kristina Jevdokimenko,
  • Ronja Rehm,
  • Jannik Hentze,
  • Paola Agüi-Gonzalez,
  • Momchil Ninov,
  • Erik Maier,
  • Yin Wu,
  • Felix Lange,
  • Agata Witkowska,
  • Svenja Bolz,
  • Francesca Pennacchietti,
  • Martina Damenti,
  • Natalie Kaempf,
  • Vladimir Khayenko,
  • Carles Calatayud,
  • Viveka Nand Malviya,
  • Natali L. Chanaday,
  • Emma Scaletti Hutchinson,
  • Hao Liu,
  • Kirsten Weyand,
  • Valentin Schwarze,
  • Daniela Ivanova,
  • Tristan P. Wallis,
  • Christopher Small,
  • Hans M. Maric,
  • Merja Joensuu,
  • Michael A. Cousin,
  • Frédéric A. Meunier,
  • Patrik Verstreken,
  • Ilaria Testa,
  • Ege T. Kavalali,
  • Volker Haucke,
  • Stefan Jakobs,
  • Henning Urlaub,
  • Nils Brose,
  • Benjamin H. Cooper,
  • Pål Stenmark,
  • Felipe Opazo,
  • Reinhard Jahn,
  • Silvio O. Rizzoli,
  • Eugenio F. Fornasiero

摘要

Synaptic neurotransmission is a critical hallmark of brain activity and one of the first processes affected in neural diseases. Monitoring this process, particularly synaptic vesicle recycling, in living cells has been instrumental in revealing the mechanisms responsible for neurotransmitter release. However, currently available reporters suffer from limitations, such as large probe sizes or limited compatibility for human neurons, hampering the quantitative analysis of synaptic pathophysiology. Here, we describe the NbLumSyt1 toolkit, a panel of nanobody-based affinity probes that target the luminal domain of the synaptic vesicle protein Synaptotagmin 1 (Syt1). These new tools enable quantitative, noninvasive imaging and functional interrogation of Syt1 exo-endocytosis and trafficking in human neurons, with unprecedented precision, versatility and cost efficiency, in technologies ranging from fixed- and live-cell super-resolution imaging to electron microscopy and mass spectrometry. Overall, NbLumSyt1 nanobinders provide a valuable platform for studying synaptic physiology and pathophysiology, benefiting fundamental neuroscience and translational efforts to study and develop treatments for brain-related disorders.

Graphical Abstract