Improved methods for marking active neuron populations

Benjamien Moeyaert, Graham Holt, Rajtarun Madangopal, Alberto Perez-Alvarez, Brenna C. Fearey, Nicholas F. Trojanowski, Julia Ledderose, Timothy A. Zolnik, Aniruddha Das, Davina Patel, Timothy A. Brown, Robert N. S. Sachdev, Britta J. Eickholt, Matthew E. Larkum, Gina G. Turrigiano, Hod Dana, Christine E. Gee, Thomas G. Oertner, Bruce T. Hope and Eric R. Schreiter ()
Additional contact information
Benjamien Moeyaert: Howard Hughes Medical Institute
Graham Holt: Howard Hughes Medical Institute
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
Matthew E. Larkum: Humboldt University
Gina G. Turrigiano: Brandeis University
Hod Dana: Howard Hughes Medical Institute
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

Nature Communications, 2018, vol. 9, issue 1, 1-12

Abstract: 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.

Date: 2018
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DOI: 10.1038/s41467-018-06935-2

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