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Protocadherin-1 Localization and Cell-Adhesion Function in Airway Epithelial Cells in Asthma
BACKGROUND: The asthma gene PCDH1 encodes Protocadherin-1, a putative adhesion molecule of unknown function expressed in the airway epithelium. Here, we characterize the localization, differential expression, homotypic adhesion specificity and function of PCDH1 in airway epithelial cells in asthma....
Autores principales: | , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5049773/ https://www.ncbi.nlm.nih.gov/pubmed/27701444 http://dx.doi.org/10.1371/journal.pone.0163967 |
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author | Faura Tellez, Grissel Willemse, Brigitte W. M. Brouwer, Uilke Nijboer-Brinksma, Susan Vandepoele, Karl Noordhoek, Jacobien A. Heijink, Irene de Vries, Maaike Smithers, Natalie P. Postma, Dirkje S. Timens, Wim Wiffen, Laura van Roy, Frans Holloway, John W. Lackie, Peter M. Nawijn, Martijn C. Koppelman, Gerard H. |
author_facet | Faura Tellez, Grissel Willemse, Brigitte W. M. Brouwer, Uilke Nijboer-Brinksma, Susan Vandepoele, Karl Noordhoek, Jacobien A. Heijink, Irene de Vries, Maaike Smithers, Natalie P. Postma, Dirkje S. Timens, Wim Wiffen, Laura van Roy, Frans Holloway, John W. Lackie, Peter M. Nawijn, Martijn C. Koppelman, Gerard H. |
author_sort | Faura Tellez, Grissel |
collection | PubMed |
description | BACKGROUND: The asthma gene PCDH1 encodes Protocadherin-1, a putative adhesion molecule of unknown function expressed in the airway epithelium. Here, we characterize the localization, differential expression, homotypic adhesion specificity and function of PCDH1 in airway epithelial cells in asthma. METHODS: We performed confocal fluorescence microscopy to determine subcellular localization of PCDH1 in 16HBE cells and primary bronchial epithelial cells (PBECs) grown at air-liquid interface. Next, to compare PCDH1 expression and localization in asthma and controls we performed qRT-PCR and fluorescence microscopy in PBECs and immunohistochemistry on airway wall biopsies. We examined homotypic adhesion specificity of HEK293T clones overexpressing fluorescently tagged-PCDH1 isoforms. Finally, to evaluate the role for PCDH1 in epithelial barrier formation and repair, we performed siRNA knockdown-studies and measured epithelial resistance. RESULTS: PCDH1 localized to the cell membrane at cell-cell contact sites, baso-lateral to adherens junctions, with increasing expression during epithelial differentiation. No differences in gene expression or localization of PCDH1 isoforms expressing the extracellular domain were observed in either PBECs or airway wall biopsies between asthma patients and controls. Overexpression of PCDH1 mediated homotypic interaction, whereas downregulation of PCDH1 reduced epithelial barrier formation, and impaired repair after wounding. CONCLUSIONS: In conclusion, PCDH1 is localized to the cell membrane of bronchial epithelial cells baso-lateral to the adherens junction. Expression of PCDH1 is not reduced nor delocalized in asthma even though PCDH1 contributes to homotypic adhesion, epithelial barrier formation and repair. |
format | Online Article Text |
id | pubmed-5049773 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-50497732016-10-27 Protocadherin-1 Localization and Cell-Adhesion Function in Airway Epithelial Cells in Asthma Faura Tellez, Grissel Willemse, Brigitte W. M. Brouwer, Uilke Nijboer-Brinksma, Susan Vandepoele, Karl Noordhoek, Jacobien A. Heijink, Irene de Vries, Maaike Smithers, Natalie P. Postma, Dirkje S. Timens, Wim Wiffen, Laura van Roy, Frans Holloway, John W. Lackie, Peter M. Nawijn, Martijn C. Koppelman, Gerard H. PLoS One Research Article BACKGROUND: The asthma gene PCDH1 encodes Protocadherin-1, a putative adhesion molecule of unknown function expressed in the airway epithelium. Here, we characterize the localization, differential expression, homotypic adhesion specificity and function of PCDH1 in airway epithelial cells in asthma. METHODS: We performed confocal fluorescence microscopy to determine subcellular localization of PCDH1 in 16HBE cells and primary bronchial epithelial cells (PBECs) grown at air-liquid interface. Next, to compare PCDH1 expression and localization in asthma and controls we performed qRT-PCR and fluorescence microscopy in PBECs and immunohistochemistry on airway wall biopsies. We examined homotypic adhesion specificity of HEK293T clones overexpressing fluorescently tagged-PCDH1 isoforms. Finally, to evaluate the role for PCDH1 in epithelial barrier formation and repair, we performed siRNA knockdown-studies and measured epithelial resistance. RESULTS: PCDH1 localized to the cell membrane at cell-cell contact sites, baso-lateral to adherens junctions, with increasing expression during epithelial differentiation. No differences in gene expression or localization of PCDH1 isoforms expressing the extracellular domain were observed in either PBECs or airway wall biopsies between asthma patients and controls. Overexpression of PCDH1 mediated homotypic interaction, whereas downregulation of PCDH1 reduced epithelial barrier formation, and impaired repair after wounding. CONCLUSIONS: In conclusion, PCDH1 is localized to the cell membrane of bronchial epithelial cells baso-lateral to the adherens junction. Expression of PCDH1 is not reduced nor delocalized in asthma even though PCDH1 contributes to homotypic adhesion, epithelial barrier formation and repair. Public Library of Science 2016-10-04 /pmc/articles/PMC5049773/ /pubmed/27701444 http://dx.doi.org/10.1371/journal.pone.0163967 Text en © 2016 Faura Tellez et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Faura Tellez, Grissel Willemse, Brigitte W. M. Brouwer, Uilke Nijboer-Brinksma, Susan Vandepoele, Karl Noordhoek, Jacobien A. Heijink, Irene de Vries, Maaike Smithers, Natalie P. Postma, Dirkje S. Timens, Wim Wiffen, Laura van Roy, Frans Holloway, John W. Lackie, Peter M. Nawijn, Martijn C. Koppelman, Gerard H. Protocadherin-1 Localization and Cell-Adhesion Function in Airway Epithelial Cells in Asthma |
title | Protocadherin-1 Localization and Cell-Adhesion Function in Airway Epithelial Cells in Asthma |
title_full | Protocadherin-1 Localization and Cell-Adhesion Function in Airway Epithelial Cells in Asthma |
title_fullStr | Protocadherin-1 Localization and Cell-Adhesion Function in Airway Epithelial Cells in Asthma |
title_full_unstemmed | Protocadherin-1 Localization and Cell-Adhesion Function in Airway Epithelial Cells in Asthma |
title_short | Protocadherin-1 Localization and Cell-Adhesion Function in Airway Epithelial Cells in Asthma |
title_sort | protocadherin-1 localization and cell-adhesion function in airway epithelial cells in asthma |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5049773/ https://www.ncbi.nlm.nih.gov/pubmed/27701444 http://dx.doi.org/10.1371/journal.pone.0163967 |
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