Tags

Type your tag names separated by a space and hit enter

Observing planar cell polarity in multiciliated mouse airway epithelial cells.
Methods Cell Biol. 2015; 127:37-54.MC

Abstract

The concerted movement of cilia propels inhaled contaminants out of the lungs, safeguarding the respiratory system from toxins, pathogens, pollutants, and allergens. Motile cilia on the multiciliated cells (MCCs) of the airway epithelium are physically oriented along the tissue axis for directional motility, which depends on the planar cell polarity (PCP) signaling pathway. The MCCs of the mouse respiratory epithelium have emerged as an important model for the study of motile ciliogenesis and the PCP signaling mechanism. Unlike other motile ciliated or planar polarized tissues, airway epithelial cells are relatively easily accessible and primary cultures faithfully model many of the essential features of the in vivo tissue. There is growing interest in understanding how cells acquire and polarize motile cilia due to the impact of mucociliary clearance on respiratory health. Here, we present methods for observing and quantifying the planar polarized orientation of motile cilia both in vivo and in primary culture airway epithelial cells. We describe how to acquire and evaluate electron and light microscopy images of ciliary ultrastructural features that reveal planar polarized orientation. Furthermore, we describe the immunofluorescence localization of PCP pathway components as a simple readout for airway epithelial planar polarization and ciliary orientation. These methods can be adapted to observe ciliary orientation in other multi- and monociliated cells and to detect PCP pathway activity in any tissue or cell type.

Authors+Show Affiliations

Department of Pathology, Stanford University School of Medicine, Stanford, CA, USA.Department of Biology, Stanford University, Stanford, CA, USA.Department of Biology, Stanford University, Stanford, CA, USA; Department of Genetics, Stanford University School of Medicine, Stanford, CA, USA.Department of Pathology, Stanford University School of Medicine, Stanford, CA, USA.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

25837385

Citation

Vladar, Eszter K., et al. "Observing Planar Cell Polarity in Multiciliated Mouse Airway Epithelial Cells." Methods in Cell Biology, vol. 127, 2015, pp. 37-54.
Vladar EK, Lee YL, Stearns T, et al. Observing planar cell polarity in multiciliated mouse airway epithelial cells. Methods Cell Biol. 2015;127:37-54.
Vladar, E. K., Lee, Y. L., Stearns, T., & Axelrod, J. D. (2015). Observing planar cell polarity in multiciliated mouse airway epithelial cells. Methods in Cell Biology, 127, 37-54. https://doi.org/10.1016/bs.mcb.2015.01.016
Vladar EK, et al. Observing Planar Cell Polarity in Multiciliated Mouse Airway Epithelial Cells. Methods Cell Biol. 2015;127:37-54. PubMed PMID: 25837385.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Observing planar cell polarity in multiciliated mouse airway epithelial cells. AU - Vladar,Eszter K, AU - Lee,Yin Loon, AU - Stearns,Tim, AU - Axelrod,Jeffrey D, Y1 - 2015/03/07/ PY - 2015/4/4/entrez PY - 2015/4/4/pubmed PY - 2016/1/27/medline KW - Ciliogenesis KW - De novo basal body formation KW - Inton retaining transcripts KW - Marsilea KW - Post-transcriptional regulation KW - Rapid development KW - Spermatogenesis SP - 37 EP - 54 JF - Methods in cell biology JO - Methods Cell Biol. VL - 127 N2 - The concerted movement of cilia propels inhaled contaminants out of the lungs, safeguarding the respiratory system from toxins, pathogens, pollutants, and allergens. Motile cilia on the multiciliated cells (MCCs) of the airway epithelium are physically oriented along the tissue axis for directional motility, which depends on the planar cell polarity (PCP) signaling pathway. The MCCs of the mouse respiratory epithelium have emerged as an important model for the study of motile ciliogenesis and the PCP signaling mechanism. Unlike other motile ciliated or planar polarized tissues, airway epithelial cells are relatively easily accessible and primary cultures faithfully model many of the essential features of the in vivo tissue. There is growing interest in understanding how cells acquire and polarize motile cilia due to the impact of mucociliary clearance on respiratory health. Here, we present methods for observing and quantifying the planar polarized orientation of motile cilia both in vivo and in primary culture airway epithelial cells. We describe how to acquire and evaluate electron and light microscopy images of ciliary ultrastructural features that reveal planar polarized orientation. Furthermore, we describe the immunofluorescence localization of PCP pathway components as a simple readout for airway epithelial planar polarization and ciliary orientation. These methods can be adapted to observe ciliary orientation in other multi- and monociliated cells and to detect PCP pathway activity in any tissue or cell type. SN - 0091-679X UR - https://www.unboundmedicine.com/medline/citation/25837385/Observing_planar_cell_polarity_in_multiciliated_mouse_airway_epithelial_cells_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0091-679X(15)00036-9 DB - PRIME DP - Unbound Medicine ER -