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Mass Synaptometry: Applying Mass Cytometry to Single Synapse Analysis

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Synaptic Vesicles

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2417))

Abstract

Synaptic degeneration is one of the earliest and phenotypically most significant features associated with numerous neurodegenerative conditions, including Alzheimer’s and Parkinson’s diseases. Synaptic changes are also known to be important in neurocognitive disorders such as schizophrenia and autism spectrum disorders. Several labs, including ours, have demonstrated that conventional (fluorescence-based) flow cytometry of individual synaptosomes is a robust and reproducible method. However, the repertoire of probes needed to assess comprehensively the type of synapse, pathologic proteins (including protein products of risk genes discovered in GWAS), and markers of stress and injury far exceeds what is achievable with conventional flow cytometry. We recently developed a method that applies CyTOF (Cytometry by Time-Of-Flight mass spectrometry) to high-dimensional analysis of individual human synaptosomes, overcoming many of the multiplexing limitations of conventional flow cytometry. We call this new method Mass Synaptometry. Here we describe the preparation of synaptosomes from human and mouse brain, the generation and quality control of the “SynTOF” (Synapse by Time-Of-Flight mass spectrometry) antibody panel, the staining protocol, and CyTOF parameter setup for acquisition, post-acquisition processing, and analysis.

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Acknowledgments

This work was supported by grants from the NIH: P50 NS062684, P50 AG047366, RF1 AG053959, R01 AG056287, R01AG057915, DP2 EB024246, and P50 AG005136 (CDK), and by the Nancy and Buster Alvord Endowment. We thank M. Holden and M. Leipold of the Stanford Human Immune Monitoring Core for their assistance and guidance, A. Beller from the Department of Pathology, University of Washington for administrative support, and E. Fox from Department of Pathology, Stanford University for helpful discussions.

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Authors and Affiliations

  1. Department of Pathology, Stanford University Medical Center, Stanford, CA, USA

    Chandresh R. Gajera, Rosemary Fernandez, Kathleen S. Montine, Sean C. Bendall & Thomas J. Montine

  2. Department of Pathology, University of Washington, Seattle, WA, USA

    Nadia Postupna & C. Dirk Keene

Authors
  1. Chandresh R. Gajera
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  2. Rosemary Fernandez
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  3. Nadia Postupna
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  4. Kathleen S. Montine
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  5. C. Dirk Keene
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  6. Sean C. Bendall
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  7. Thomas J. Montine
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Corresponding author

Correspondence to Chandresh R. Gajera .

Editor information

Editors and Affiliations

  1. Department of Psychiatry and Psychotherapy, Universitätsklinikum Erlangen, Erlangen, Bayern, Germany

    Jana Dahlmanns

  2. Institute for Physiology and Pathophysiology, University of Erlangen-Nuremberg, Erlangen, Germany

    Marc Dahlmanns

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Gajera, C.R. et al. (2022). Mass Synaptometry: Applying Mass Cytometry to Single Synapse Analysis. In: Dahlmanns, J., Dahlmanns, M. (eds) Synaptic Vesicles. Methods in Molecular Biology, vol 2417. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1916-2_6

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  • DOI: https://doi.org/10.1007/978-1-0716-1916-2_6

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1915-5

  • Online ISBN: 978-1-0716-1916-2

  • eBook Packages: Springer Protocols

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