PCDH1
| Autor: | Irene Hejink, John W. Holloway, Brigitte W. M. Willemse, de Maaike Vries, Martijn C. Nawijn, Grissel Faura Tellez, Dirkje S. Postma, Jacobien A. Noordhoek, Natalie Smithers, Wim Timens, Peter M. Lackie, Gerard H. Koppelman, Laura Wiffen, Uilke Brouwer |
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| Přispěvatelé: | Groningen Research Institute for Asthma and COPD (GRIAC) |
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
medicine.diagnostic_test
business.industry Transfection Immunofluorescence Occludin Epithelium respiratory tract diseases Cell biology Cell membrane Adherens junction Epithelial cell medicine.anatomical_structure Asthma - mechanism medicine Respiratory epithelium Experimental approaches Cell adhesion business |
| Zdroj: | European Respiratory Journal, 46. EUROPEAN RESPIRATORY SOC JOURNALS LTD |
| ISSN: | 0903-1936 |
| Popis: | Protocadherin-1 (PCDH1) genetic variants have been associated with bronchial hyperresponsiveness and asthma suggesting a role in asthma pathogenesis. This asthma gene encodes a putative adhesion molecule of unknown function. We investigated the localization, differential expression in airway epithelial cells in asthma and role of PCDH1 as an adhesion molecule. 16HBE cells were transfected with constructs encoding fluorescently tagged PCDH1 isoforms. Endogenous PCDH1 was detected in 16HBE cells and during differentiation of primary bronchial epithelial cells (PBECs). PBECs were dual-stained with PCDH1 and E-cadherin or Occludin. Immunohistochemistry on airway wall biopsies from asthma and controls subjects as well as qRT-PCR and immunofluorescence on PBECs were performed. Finally, we knocked down PCDH1 in 16HBE cells and measured epithelial resistance using electric cell substrate impedance sensing. Fluorescent microscopy of 16HBE cellsrevealed membrane localization of PCDH1 isoforms. Confocal microscopy demonstrated PCDH1 at cell-cell contact sites, basolaterally to E-cadherin and basal to Occludin, with increasing expression towards the apical surface during epithelial differentiation. Airway wall biopsies and PBECs from asthmatics and controls showed no difference in either PCDH1 expression or localization. Downregulation of PCDH1 decreased epithelial barrier formation, and impaired repair after wounding. In conclusion, PCDH1 isoforms localized to the cell membrane. PCDH1 is located mainly apical but basal to the adherens junctions in differentiated airway epithelial cells. Expression of extracellular PCDH1 is not reduced nor delocalized in asthma. PCDH1 maintains epithelial barrier formation and repair. |
| Databáze: | OpenAIRE |
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