Abstract

Dysfunctional and leaky blood vessels resulting from disruption of the endothelial cell (EC) barrier accompanies numerous diseases. The EC barrier is established through endothelial cell tight and adherens junctions. However, the expression patterning and precise contribution of different junctional proteins to the EC barrier is poorly understood. Here, we focus on organs with continuous endothelium to identify structural and functional in vivo characteristics of the EC barrier. Assembly of multiple single-cell RNAseq datasets into a single integrated database revealed the variability and commonalities of EC barrier patterning. Across tissues, Claudin5 exhibited diminishing expression along the arteriovenous axis, correlating with EC barrier integrity. Functional analysis identified tissue-specific differences in leakage patterning and response to the leakage agonist histamine. Loss of Claudin5 enhanced histamine-induced leakage in an organotypic and vessel type-specific manner in an inducible, EC-specific, knock-out mouse. Mechanistically, Claudin5 loss left junction ultrastructure unaffected but altered its composition, with concomitant loss of zonula occludens-1 and upregulation of VE-Cadherin expression. These findings uncover the organ-specific organisation of the EC barrier and distinct importance of Claudin5 in different vascular beds, providing insights to modify EC barrier stability in a targeted, organ-specific manner.

Data availability

The murine ear skin data has been deposited in GEO under accession number GSE202290. Further details regarding specifics of the analysis will be available upon reasonable request.

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Mark Richards

    Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    For correspondence
    mark.richards@igp.uu.se
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2266-3329
  2. Emmanuel Nwadozi

    Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    Competing interests
    The authors declare that no competing interests exist.
  3. Sagnik Pal

    Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5562-1555
  4. Pernilla Martinsson

    Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    Competing interests
    The authors declare that no competing interests exist.
  5. Mika Kaakinen

    Faculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, Finland
    Competing interests
    The authors declare that no competing interests exist.
  6. Marleen Gloger

    Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3319-7642
  7. Elin Sjöberg

    Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8799-4874
  8. Katarzyna Koltowska

    Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6841-8900
  9. Christer Betsholtz

    Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    Competing interests
    The authors declare that no competing interests exist.
  10. Lauri Eklund

    Faculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, Finland
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3177-7504
  11. Sofia Nordling

    Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    Competing interests
    The authors declare that no competing interests exist.
  12. Lena Claesson-Welsh

    Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    For correspondence
    lena.welsh@igp.uu.se
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4275-2000

Funding

Vetenskapsrådet (2020-01349)

  • Lena Claesson-Welsh

Åke Wiberg Stiftelse (M21-0109)

  • Sofia Nordling

Svenska Sällskapet för Medicinsk Forskning

  • Elin Sjöberg

Svenska Sällskapet för Medicinsk Forskning (201912)

  • Mark Richards

European Molecular Biology Organization (ALTF 923-2016)

  • Mark Richards

Knut och Alice Wallenbergs Stiftelse (2020.0057)

  • Lena Claesson-Welsh

Knut och Alice Wallenbergs Stiftelse (2019.0276)

  • Lena Claesson-Welsh

Fondation Leducq (17 CVD 03)

  • Lena Claesson-Welsh

Cancerfonden (19 0119 Pj)

  • Lena Claesson-Welsh

Cancerfonden (19 0118 Us)

  • Lena Claesson-Welsh

Cancerfonden (20 1086 Pj)

  • Marleen Gloger
  • Katarzyna Koltowska

Knut och Alice Wallenbergs Stiftelse (2017.0144)

  • Katarzyna Koltowska

Ragnar Söderbergs stiftelse (M13/17)

  • Katarzyna Koltowska

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

Reviewing Editor

  1. Gou Young Koh, Institute of Basic Science and Korea Advanced Institute of Science and Technology (KAIST), Korea (South), Republic of

Ethics

Animal experimentation: In vivo animal experiments were carried out in accordance with the ethical permit provided by the Committee on the Ethics of Animal Experiments of the University of Uppsala (permit 6789/18).

Version history

  1. Received: March 10, 2022
  2. Preprint posted: March 16, 2022 (view preprint)
  3. Accepted: July 20, 2022
  4. Accepted Manuscript published: July 21, 2022 (version 1)
  5. Version of Record published: August 3, 2022 (version 2)

Copyright

© 2022, Richards 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. Mark Richards
  2. Emmanuel Nwadozi
  3. Sagnik Pal
  4. Pernilla Martinsson
  5. Mika Kaakinen
  6. Marleen Gloger
  7. Elin Sjöberg
  8. Katarzyna Koltowska
  9. Christer Betsholtz
  10. Lauri Eklund
  11. Sofia Nordling
  12. Lena Claesson-Welsh
(2022)
Claudin5 protects the peripheral endothelial barrier in an organ and vessel type-specific manner
eLife 11:e78517.
https://doi.org/10.7554/eLife.78517

Share this article

https://doi.org/10.7554/eLife.78517

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