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Improved methods for marking active neuron populations

Author

Listed:
  • Benjamien Moeyaert

    (Howard Hughes Medical Institute)

  • Graham Holt

    (Howard Hughes Medical Institute
    Duke University)

  • Rajtarun Madangopal

    (National Institute on Drug Abuse, National Institutes of Health)

  • Alberto Perez-Alvarez

    (University Medical Center Hamburg-Eppendorf)

  • Brenna C. Fearey

    (University Medical Center Hamburg-Eppendorf)

  • Nicholas F. Trojanowski

    (Brandeis University)

  • Julia Ledderose

    (Charité - Universitätsmedizin Berlin)

  • Timothy A. Zolnik

    (Humboldt University)

  • Aniruddha Das

    (Cleveland Clinic Foundation)

  • Davina Patel

    (Cleveland Clinic Foundation)

  • Timothy A. Brown

    (Howard Hughes Medical Institute)

  • Robert N. S. Sachdev

    (Humboldt University)

  • Britta J. Eickholt

    (Charité - Universitätsmedizin Berlin
    Humboldt University)

  • Matthew E. Larkum

    (Humboldt University)

  • Gina G. Turrigiano

    (Brandeis University)

  • Hod Dana

    (Howard Hughes Medical Institute
    Cleveland Clinic Foundation)

  • Christine E. Gee

    (University Medical Center Hamburg-Eppendorf)

  • Thomas G. Oertner

    (University Medical Center Hamburg-Eppendorf)

  • Bruce T. Hope

    (National Institute on Drug Abuse, National Institutes of Health)

  • Eric R. Schreiter

    (Howard Hughes Medical Institute)

Abstract

Marking functionally distinct neuronal ensembles with high spatiotemporal resolution is a key challenge in systems neuroscience. We recently introduced CaMPARI, an engineered fluorescent protein whose green-to-red photoconversion depends on simultaneous light exposure and elevated calcium, which enabled marking active neuronal populations with single-cell and subsecond resolution. However, CaMPARI (CaMPARI1) has several drawbacks, including background photoconversion in low calcium, slow kinetics and reduced fluorescence after chemical fixation. In this work, we develop CaMPARI2, an improved sensor with brighter green and red fluorescence, faster calcium unbinding kinetics and decreased photoconversion in low calcium conditions. We demonstrate the improved performance of CaMPARI2 in mammalian neurons and in vivo in larval zebrafish brain and mouse visual cortex. Additionally, we herein develop an immunohistochemical detection method for specific labeling of the photoconverted red form of CaMPARI. The anti-CaMPARI-red antibody provides strong labeling that is selective for photoconverted CaMPARI in activated neurons in rodent brain tissue.

Suggested Citation

  • Benjamien Moeyaert & Graham Holt & Rajtarun Madangopal & Alberto Perez-Alvarez & Brenna C. Fearey & Nicholas F. Trojanowski & Julia Ledderose & Timothy A. Zolnik & Aniruddha Das & Davina Patel & Timot, 2018. "Improved methods for marking active neuron populations," Nature Communications, Nature, vol. 9(1), pages 1-12, December.
    • Handle: RePEc:nat:natcom:v:9:y:2018:i:1:d:10.1038_s41467-018-06935-2
    DOI: 10.1038/s41467-018-06935-2
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    Citations

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    Cited by:

    1. Irene Serra & Julio Esparza & Laura Delgado & Cristina Martín-Monteagudo & Margalida Puigròs & Petar Podlesniy & Ramón Trullás & Marta Navarrete, 2022. "Ca2+-modulated photoactivatable imaging reveals neuron-astrocyte glutamatergic circuitries within the nucleus accumbens," Nature Communications, Nature, vol. 13(1), pages 1-18, December.
    2. Yung-Tian A. Gau & Eric T. Hsu & Richard J. Cha & Rebecca W. Pak & Loren L. Looger & Jin U. Kang & Dwight E. Bergles, 2024. "Multicore fiber optic imaging reveals that astrocyte calcium activity in the mouse cerebral cortex is modulated by internal motivational state," Nature Communications, Nature, vol. 15(1), pages 1-21, December.
    3. Aniruddha Das & Sarah Holden & Julie Borovicka & Jacob Icardi & Abigail O’Niel & Ariel Chaklai & Davina Patel & Rushik Patel & Stefanie Kaech Petrie & Jacob Raber & Hod Dana, 2023. "Large-scale recording of neuronal activity in freely-moving mice at cellular resolution," Nature Communications, Nature, vol. 14(1), pages 1-12, December.
    4. Franziska Bierbuesse & Anaïs C. Bourges & Vincent Gielen & Viola Mönkemöller & Wim Vandenberg & Yi Shen & Johan Hofkens & Pieter Vanden Berghe & Robert E. Campbell & Benjamien Moeyaert & Peter Dedecke, 2022. "Absolute measurement of cellular activities using photochromic single-fluorophore biosensors and intermittent quantification," Nature Communications, Nature, vol. 13(1), pages 1-13, December.
    5. Paul J. Lamothe-Molina & Andreas Franzelin & Lennart Beck & Dong Li & Lea Auksutat & Tim Fieblinger & Laura Laprell & Joachim Alhbeck & Christine E. Gee & Matthias Kneussel & Andreas K. Engel & Claus , 2022. "ΔFosB accumulation in hippocampal granule cells drives cFos pattern separation during spatial learning," Nature Communications, Nature, vol. 13(1), pages 1-16, December.

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