1. Neuroscience

Rapid purification and metabolomic profiling of synaptic vesicles from mammalian brain

  1. Lynne Chantranupong
  2. Jessica L Saulnier
  3. Wengang Wang
  4. Drew R Jones
  5. Michael E Pacold
  6. Bernardo L Sabatini  Is a corresponding author
  1. Harvard Medical School, United States
  2. NYU Langone Health, United States
  3. Howard Hughes Medical Institute, Harvard Medical School, United States
https://doi.org/10.7554/eLife.59699
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Cite this article
  1. Lynne Chantranupong
  2. Jessica L Saulnier
  3. Wengang Wang
  4. Drew R Jones
  5. Michael E Pacold
  6. Bernardo L Sabatini
(2020)
Rapid purification and metabolomic profiling of synaptic vesicles from mammalian brain
eLife 9:e59699.
https://doi.org/10.7554/eLife.59699
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Abstract

Neurons communicate by the activity-dependent release of small-molecule neurotransmitters packaged into synaptic vesicles (SVs). Although many molecules have been identified as neurotransmitters, technical limitations have precluded a full metabolomic analysis of synaptic vesicle content. Here, we present a workflow to rapidly isolate SVs and to interrogate their metabolic contents at high-resolution using mass spectrometry. We validated the enrichment of glutamate in SVs of primary cortical neurons using targeted polar metabolomics. Unbiased and extensive global profiling of SVs isolated from these neurons revealed that the only detectable polar metabolites they contain are the established neurotransmitters glutamate and GABA. In addition, we adapted the approach to enable quick capture of SVs directly from brain tissue and determined the neurotransmitter profiles of diverse brain regions in a cell-type specific manner. The speed, robustness, and precision of this method to interrogate SV contents will facilitate novel insights into the chemical basis of neurotransmission.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for all figures.

Article and author information

Author details

  1. Lynne Chantranupong

    Neurobiology, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.

  2. Jessica L Saulnier

    Neurobiology, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.

  3. Wengang Wang

    Neurobiology, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.

  4. Drew R Jones

    Metabolomics Core Resource Laboratory, NYU Langone Health, New York, United States
    Competing interests
    The authors declare that no competing interests exist.

  5. Michael E Pacold

    Radiation Oncology, NYU Langone Health, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3688-2378

  6. Bernardo L Sabatini

    Department of Neurobiology, Howard Hughes Medical Institute, Harvard Medical School, Boston, United States
    For correspondence
    bsabatini@hms.harvard.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0095-9177

Funding

Howard Hughes Medical Institute (Investigator)

  • Bernardo L Sabatini

National Institute of Neurological Disorders and Stroke (R37NS046579)

  • Bernardo L Sabatini

Howard Hughes Medical Institute (Hanna Gray Fellowship)

  • Lynne Chantranupong

Mary Kay Foundation (Cancer Research Grant 017-032)

  • Michael E Pacold

Hearst Foundation (V Foundation V Scholar Grant (V2017-004))

  • Michael E Pacold

National Cancer Institute (K22 Career Transition Award (1K22CA212059))

  • Michael E Pacold

NIH (R01 NS108151-01)

  • Drew R Jones

FNIH (RFA 2018-PACT001)

  • Drew R Jones

NIH (HHS-NIH-NIAD-BAA2018)

  • Drew R Jones

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All experimental manipulations were performed in accordance with protocols (#03551) approved by the Harvard Standing Committee on Animal Care following guidelines described in the US National Institutes of Health Guide for the Care and Use of Laboratory Animals.

Copyright

© 2020, Chantranupong et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Lynne Chantranupong
  2. Jessica L Saulnier
  3. Wengang Wang
  4. Drew R Jones
  5. Michael E Pacold
  6. Bernardo L Sabatini
(2020)
Rapid purification and metabolomic profiling of synaptic vesicles from mammalian brain
eLife 9:e59699.
https://doi.org/10.7554/eLife.59699

Share this article

https://doi.org/10.7554/eLife.59699

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