Amyloid-β Peptides Disrupt Interactions between VAMP-2 and SNAP-25 in Neuronal Cells as Determined by FRET/FLIM

Nidhi Sharda, Thomas Pengo, Zengtao Wang, Karunya K. Kandimalla, Ottavio Arancio

Research output: Contribution to journalArticlepeer-review


Synaptic dysfunction prevalent in Alzheimer's disease (AD) brain is closely associated with increased accumulation of amyloid-β (Aβ) peptides in the brain parenchyma. It is widely believed that Aβ peptides trigger synaptic dysfunction by interfering with the synaptic vesicular fusion and the release of neurotransmitters, primarily facilitated by the SNARE protein complexes formed by VAMP-2, SNAP-25, and syntaxin-1. However, Aβ interactions with SNARE proteins to ultimately disrupt synaptic vesicular fusion are not well understood. Objective: Our objective is to elucidate mechanisms by which Aβ peptides perturb SNARE complexes. Methods: Intensity (qualitative) and lifetime (quantitative) based measurements involving Forster (fluorescence) resonance energy transfer (FRET) followed by fluorescence lifetime imaging microscopy (FLIM) were employed to investigate the effect of Aβ peptides on dynamic interactions between VAMP-2, labeled with cerulean (Cer) at the N-terminus (FRET donor), and SNAP-25 labeled with citrine (Cit) on the N-terminus (FRET acceptor). The FRET and FLIM interactions at the exocytosis locations on the pre-synaptic membrane were recorded under spontaneous and high potassium evoked conditions. Moreover, cellular accumulation of fluorescein labeled Aβ (F-Aβ) peptides and their co-localization with Cer-VAMP2 was investigated by confocal microscopy. Results: The F-Aβ40 and F-Aβ42 are internalized by differentiated N2A cells, where they colocalize with Cer-VAMP2. Both Aβ40 and Aβ42 decrease interactions between the N-termini of Cer-VAMP2 and Cit-SNAP25 in N2A cells, as determined by FRET/FLIM. Conclusion: By perturbing the N-terminal interactions between VAMP-2 and SNAP-25, Aβ40 and Aβ42, can directly interfere with the SNARE complex formation, which is critical for the docking and fusion of synaptic vesicles.

Original languageEnglish (US)
Pages (from-to)423-435
Number of pages13
JournalJournal of Alzheimer's Disease
Issue number1
StatePublished - 2020

Bibliographical note

Funding Information:
In conclusion, these findings provide first direct evidence of the perturbations caused by Aβ peptides to the N-termini interactions of SNAP-25 and The authors thank Dr. Robert Zucker (Department of Molecular and Cellular Biology, University of California at Berkeley) for kindly donating plasmid mCer-4aa-VAMP2 (Addgene plasmid #53233) and its corresponding FRET partner mCit-4aa-SNAP25B (Addgene plasmid #53235) as kind gifts. Additionally, the authors thank the University of Minnesota, University Imaging Centers for experimental and technical support. Finally, the authors acknowledge the funding support and assistance from the Minnesota Partnership grant MNP# 15.31.


  • Alzheimer's disease
  • Amyloid-β peptides
  • exocytosis
  • FLIM
  • FRET
  • SNAP-25
  • SNAREs
  • synaptic vesicular fusion
  • VAMP-2

PubMed: MeSH publication types

  • Journal Article

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