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Related Concept Videos

Fusion of Secretory Vesicles with the Plasma Membrane01:26

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Proteins and neurotransmitters in secretory vesicles can be released from a cell upon vesicle docking, priming, and fusion with the plasma membrane. Vesicles are docked and primed in preparation for the quick exocytosis of their contents in response to a stimulus. The fusion process is mainly carried out by a SNAP Receptor or SNARE complex, consisting of synaptobrevin, syntaxin-1, and SNAP-25.
In 1993, Jim Rothman proposed that the antiparallel pairing of vesicular and transmembrane SNAREs, or...
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Related Experiment Video

Updated: Jun 4, 2025

Subcellular Fractionation for the Isolation of Synaptic Components from the Murine Brain
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Isolating Synaptic Vesicles from Neurospheres for Proteomics.

Caroline Brandão-Teles1, Giuliana S Zuccoli1, Marcelo Ganzella2

  • 1Laboratory of Neuroproteomics, Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas, Campinas, Brazil.

Methods in Molecular Biology (Clifton, N.J.)
|December 23, 2024
PubMed
Summary
This summary is machine-generated.

This study details a new method to isolate synaptic vesicles (SVs) from human stem cell-derived neurons. The optimized protocol enables detailed proteomic analysis for understanding neurological disorders.

Keywords:
Neural stem cellNeurospheresProteomicsSynapse vesiclesVesicle isolation

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Area of Science:

  • Neuroscience
  • Stem Cell Biology
  • Proteomics

Background:

  • Synaptic vesicles (SVs) are crucial for neuronal communication.
  • Understanding SV molecular composition is vital for neurological research.
  • Current methods for SV isolation from human induced pluripotent stem cells (hiPSCs) can be improved.

Purpose of the Study:

  • To develop and optimize a protocol for isolating synaptic vesicles (SVs) from hiPSC-derived neurospheres.
  • To enable detailed proteomic analysis of SVs for functional studies.
  • To facilitate research into neurological and neuropsychiatric disorders.

Main Methods:

  • Cultivation of hiPSC-derived neurospheres to obtain mature neurons.
  • Isolation of neurosphere-derived synaptosomes.
  • Enrichment of SVs via differential centrifugation.
  • Proteomic profiling using nano-liquid chromatography coupled with tandem mass spectrometry (nanoLC-MS/MS).

Main Results:

  • Successful isolation and enrichment of synaptic vesicles from hiPSC-derived neurospheres.
  • Generation of a detailed proteome profile of the isolated SVs.
  • Demonstration of a robust protocol for SV analysis.

Conclusions:

  • The optimized protocol provides a valuable tool for studying SV molecular heterogeneity.
  • This method can advance the understanding of neurotransmitter uptake and release mechanisms.
  • The protocol has significant potential applications in neurological and neuropsychiatric disorder research.