Re-evaluation of neuronal P2X7 expression using novel mouse models and a P2X7-specific nanobody

  1. Karina Kaczmarek-Hajek
  2. Jiong Zhang
  3. Robin Kopp
  4. Antje Grosche
  5. Björn Rissiek
  6. Anika Saul
  7. Santina Bruzzone
  8. Tobias Engel
  9. Tina Jooss
  10. Anna Krautloher
  11. Stefanie Schuster
  12. Tim Magnus
  13. Christine Stadelmann
  14. Swetlana Sirko
  15. Friedrich Koch-Nolte
  16. Volker Eulenburg
  17. Annette Nicke  Is a corresponding author
  1. Max Planck Institute for Experimental Medicine, Germany
  2. Ludwig-Maximilians-Universität München, Germany
  3. University of Regensburg, Germany
  4. University Hospital Hamburg-Eppendorf, Germany
  5. University of Genova, Italy
  6. Royal College of Surgeons in Ireland, Ireland
  7. Friedrich-Alexander University Erlangen-Nürnberg, Germany
  8. University Medical Center Göttingen, Germany

Abstract

The P2X7 channel is involved in the pathogenesis of various CNS diseases. An increasing number of studies suggest its presence in neurons where its putative functions remain controversial for more than a decade. To resolve this issue and to provide a model for analysis of P2X7 functions, we generated P2X7-BAC transgenic mice that allow visualization of functional EGFP-tagged P2X7 receptors in vivo. Extensive characterization of these mice revealed dominant P2X7-EGFP protein expression in microglia, Bergmann glia, and oligodendrocytes, but not in neurons. These findings were further validated by microglia- and oligodendrocyte-specific P2X7 deletion and a novel P2X7-specific nanobody. In addition to the first quantitative analysis of P2X7 protein expression in the CNS, we show potential consequences of its overexpression in ischemic retina and post-traumatic cerebral cortex grey matter. This novel mouse model overcomes previous limitations in P2X7 research and will help to determine its physiological roles and contribution to diseases.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files.

Article and author information

Author details

  1. Karina Kaczmarek-Hajek

    Department of Molecular Biology of Neuronal Signals, Max Planck Institute for Experimental Medicine, Göttingen, Germany
    Competing interests
    No competing interests declared.
  2. Jiong Zhang

    Department of Molecular Biology of Neuronal Signals, Max Planck Institute for Experimental Medicine, Göttingen, Germany
    Competing interests
    No competing interests declared.
  3. Robin Kopp

    Walther Straub Institute for Pharmacology and Toxicology, Ludwig-Maximilians-Universität München, Munich, Germany
    Competing interests
    No competing interests declared.
  4. Antje Grosche

    Institute for Human Genetics, University of Regensburg, Regensburg, Germany
    Competing interests
    No competing interests declared.
  5. Björn Rissiek

    Department of Neurology, University Hospital Hamburg-Eppendorf, Hamburg, Germany
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5327-5479
  6. Anika Saul

    Department of Molecular Biology of Neuronal Signals, Max Planck Institute for Experimental Medicine, Göttingen, Germany
    Competing interests
    No competing interests declared.
  7. Santina Bruzzone

    Department of Experimental Medicine, University of Genova, Genova, Italy
    Competing interests
    No competing interests declared.
  8. Tobias Engel

    Department of Physiology and Medical Physics, Royal College of Surgeons in Ireland, Dublin, Ireland
    Competing interests
    No competing interests declared.
  9. Tina Jooss

    Walther Straub Institute for Pharmacology and Toxicology, Ludwig-Maximilians-Universität München, Munich, Germany
    Competing interests
    No competing interests declared.
  10. Anna Krautloher

    Walther Straub Institute for Pharmacology and Toxicology, Ludwig-Maximilians-Universität München, Munich, Germany
    Competing interests
    No competing interests declared.
  11. Stefanie Schuster

    Institute of Biochemistry, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany
    Competing interests
    No competing interests declared.
  12. Tim Magnus

    Department of Neurology, University Hospital Hamburg-Eppendorf, Hamburg, Germany
    Competing interests
    No competing interests declared.
  13. Christine Stadelmann

    Institute of Neuropathology, University Medical Center Göttingen, Göttingen, Germany
    Competing interests
    No competing interests declared.
  14. Swetlana Sirko

    Department of Physiological Genomics, Ludwig-Maximilians-Universität München, Munich, Germany
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5950-616X
  15. Friedrich Koch-Nolte

    Department of Immunology, University Hospital Hamburg-Eppendorf, Hamburg, Germany
    Competing interests
    Friedrich Koch-Nolte, FKN receives a share of antibody sales via MediGate GmbH, a wholly owned subsidiary of the University Medical Center Hamburg-Eppendorf. FKN is a co-inventor on patent applications on P2X7-specific nanobodies (WO2010070145, WO2013178783).
  16. Volker Eulenburg

    Institute of Biochemistry, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4878-5746
  17. Annette Nicke

    Department of Molecular Biology of Neuronal Signals, Max Planck Institute for Experimental Medicine, Göttingen, Germany
    For correspondence
    annette.nicke@lrz.uni-muenchen.de
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6798-505X

Funding

Deutsche Forschungsgemeinschaft (Ni 592/4-5)

  • Annette Nicke

Deutscher Akademischer Austauschdienst

  • Santina Bruzzone

Deutsche Forschungsgemeinschaft (No 310/11-1)

  • Friedrich Koch-Nolte

Science Foundation Ireland (13/SIRG/2098)

  • Tobias Engel

Horizon 2020 Framework Programme (766124)

  • Tobias Engel
  • Annette Nicke

Deutsche Forschungsgemeinschaft (SFB 1328)

  • Tim Magnus
  • Friedrich Koch-Nolte
  • Annette Nicke

Science Foundation Ireland (17/CDA/4708)

  • Tobias Engel

Deutsche Forschungsgemeinschaft (Ni 592/4-7)

  • Annette Nicke

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

Ethics

Animal experimentation: Animal handling and experimental procedures were performed in accordance with German and European Union guidelines and were approved by the State of Upper Bavaria (stab wound injury (55.2.1.54-2532-171-11), retinal ischemia (TVV 54/12; 55.2 DMS-2532-2-182), transcardial perfusion (55.2-1-54-2532-59-2016)) and Lower Saxony (generation of BAC transgenic mice, transcardial perfusion (33.9-42502-04-12/0863), behavioral experiments (3392 42502-04-13/1123)). Status epilepticus was induced in accordance with the principles of the European Communities Council Directive (86/609/EEC) and procedures reviewed and approved by the Research Ethics Committee of the Royal College of Surgeons in Ireland (REC 205 and 1322) and performed under license from the Department of Health and Children, Ireland. All efforts were made to minimize suffering and number of animals used.

Reviewing Editor

  1. Kenton Jon Swartz, National Institute of Neurological Disorders and Stroke, National Institutes of Health, United States

Publication history

  1. Received: March 5, 2018
  2. Accepted: July 31, 2018
  3. Accepted Manuscript published: August 3, 2018 (version 1)
  4. Version of Record published: September 14, 2018 (version 2)
  5. Version of Record updated: September 19, 2018 (version 3)

Copyright

© 2018, Kaczmarek-Hajek 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. Karina Kaczmarek-Hajek
  2. Jiong Zhang
  3. Robin Kopp
  4. Antje Grosche
  5. Björn Rissiek
  6. Anika Saul
  7. Santina Bruzzone
  8. Tobias Engel
  9. Tina Jooss
  10. Anna Krautloher
  11. Stefanie Schuster
  12. Tim Magnus
  13. Christine Stadelmann
  14. Swetlana Sirko
  15. Friedrich Koch-Nolte
  16. Volker Eulenburg
  17. Annette Nicke
(2018)
Re-evaluation of neuronal P2X7 expression using novel mouse models and a P2X7-specific nanobody
eLife 7:e36217.
https://doi.org/10.7554/eLife.36217
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