1. Neuroscience

Visualizing synaptic plasticity in vivo by large-scale imaging of endogenous AMPA receptors

  1. Austin R Graves
  2. Richard H Roth
  3. Han L Tan
  4. Qianwen Zhu
  5. Alexei M Bygrave
  6. Elena Lopez-Ortega
  7. Ingie Hong
  8. Alina C Spiegel
  9. Richard C Johnson
  10. Joshua T Vogelstein
  11. Daniel J Tward
  12. Michael I Miller
  13. Richard L Huganir  Is a corresponding author
  1. Johns Hopkins University School of Medicine, United States
  2. Johns Hopkins University, United States
https://doi.org/10.7554/eLife.66809
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Cite this article
  1. Austin R Graves
  2. Richard H Roth
  3. Han L Tan
  4. Qianwen Zhu
  5. Alexei M Bygrave
  6. Elena Lopez-Ortega
  7. Ingie Hong
  8. Alina C Spiegel
  9. Richard C Johnson
  10. Joshua T Vogelstein
  11. Daniel J Tward
  12. Michael I Miller
  13. Richard L Huganir
(2021)
Visualizing synaptic plasticity in vivo by large-scale imaging of endogenous AMPA receptors
eLife 10:e66809.
https://doi.org/10.7554/eLife.66809
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Abstract

Elucidating how synaptic molecules such as AMPA receptors mediate neuronal communication and tracking their dynamic expression during behavior is crucial to understand cognition and disease, but current technological barriers preclude large-scale exploration of molecular dynamics in vivo. We have developed a suite of innovative methodologies that break through these barriers: a new knockin mouse line with fluorescently tagged endogenous AMPA receptors, two-photon imaging of hundreds of thousands of labeled synapses in behaving mice, and computer-vision-based automatic synapse detection. Using these tools, we can longitudinally track how the strength of populations of synapses changes during behavior. We used this approach to generate an unprecedentedly detailed spatiotemporal map of synapses undergoing changes in strength following sensory experience. More generally, these tools can be used as an optical probe capable of measuring functional synapse strength across entire brain areas during any behavioral paradigm, describing complex system-wide changes with molecular precision.

Data availability

We have provided Source data for our Figures and deposited the remaining data to Dryad with doi:10.5061/dryad.ttdz08m0b. All code used to analyze and process data is freely available on GitHub, with links specified in the manuscript.

The following data sets were generated
    1. Huganir RL
    2. Graves A
    (2021) Raw data for Graves et. al 2021 eLife
    Dryad Digital Repository, doi:10.5061/dryad.ttdz08m0b.
    https://doi.org/10.5061/dryad.ttdz08m0b

Article and author information

Author details

  1. Austin R Graves

    Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, United States
    Competing interests
    No competing interests declared.

  2. Richard H Roth

    Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6855-999X

  3. Han L Tan

    Solomon H Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5163-7720

  4. Qianwen Zhu

    Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, United States
    Competing interests
    No competing interests declared.

  5. Alexei M Bygrave

    Solomon H Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2291-923X

  6. Elena Lopez-Ortega

    Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, United States
    Competing interests
    No competing interests declared.

  7. Ingie Hong

    Solomon H Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7246-9233

  8. Alina C Spiegel

    Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, United States
    Competing interests
    No competing interests declared.

  9. Richard C Johnson

    Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, United States
    Competing interests
    No competing interests declared.

  10. Joshua T Vogelstein

    Biomedical Engineering, Johns Hopkins University, Baltimore, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2487-6237

  11. Daniel J Tward

    Center for Imaging Science, Johns Hopkins University, Baltimore, United States
    Competing interests
    No competing interests declared.

  12. Michael I Miller

    Center for Imaging Science, Johns Hopkins University, Baltimore, United States
    Competing interests
    Michael I Miller, Dr. Miller is a joint owner of AnatomyWorks. Dr. Miller's relationship with Anatomy-Works is being handled under full disclosure by the Johns Hopkins University..

  13. Richard L Huganir

    Solomon H Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, United States
    For correspondence
    rhuganir@jhmi.edu
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9783-5183

Funding

National Institutes of Health (R21 AG063193)

  • Austin R Graves
  • Richard L Huganir

Kavli Foundation

  • Austin R Graves
  • Alina C Spiegel
  • Daniel J Tward

National Institutes of Health (R01 MH123212)

  • Austin R Graves
  • Alexei M Bygrave
  • Michael I Miller
  • Richard L Huganir

Schmidt Science Nascent Innovation Grant (1)

  • Austin R Graves
  • Joshua T Vogelstein
  • Richard L Huganir

National Institutes of Health (K99 MH124920)

  • Alexei M Bygrave

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

Ethics

Animal experimentation: These studies were conducted in accordance with US Public Health Service on Human Care and Use of Laboratory Animals (PHS Policy) and all procedures involving animals were approved by the Johns Hopkins Animal Care and Use Committee (ACUC) protocols (MO19M274, MO20M372, MO20M92, MO20M336). All surgeries were performed under isoflourane anesthesia. Every effort was made to reduce or eliminate pain and suffering during all surgical procedures, in vivo imaging sessions, and behavioral experiments.

Copyright

© 2021, Graves 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. Austin R Graves
  2. Richard H Roth
  3. Han L Tan
  4. Qianwen Zhu
  5. Alexei M Bygrave
  6. Elena Lopez-Ortega
  7. Ingie Hong
  8. Alina C Spiegel
  9. Richard C Johnson
  10. Joshua T Vogelstein
  11. Daniel J Tward
  12. Michael I Miller
  13. Richard L Huganir
(2021)
Visualizing synaptic plasticity in vivo by large-scale imaging of endogenous AMPA receptors
eLife 10:e66809.
https://doi.org/10.7554/eLife.66809

Share this article

https://doi.org/10.7554/eLife.66809

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