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A novel method for live imaging of human airway cilia using wheat germ agglutinin

Multiciliated epithelial cells in the airway are essential for mucociliary clearance. Their function relies on coordinated, metachronal and directional ciliary beating, appropriate mucus secretion and airway surface hydration. However, current conventional methods for observing human airway ciliary...

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Autores principales: Nakamura, Ryosuke, Katsuno, Tatsuya, Kishimoto, Yo, Kaba, Shinji, Yoshimatsu, Masayoshi, Kitamura, Morimasa, Suehiro, Atsushi, Hiwatashi, Nao, Yamashita, Masaru, Tateya, Ichiro, Omori, Koichi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7468155/
https://www.ncbi.nlm.nih.gov/pubmed/32879324
http://dx.doi.org/10.1038/s41598-020-71049-z
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author Nakamura, Ryosuke
Katsuno, Tatsuya
Kishimoto, Yo
Kaba, Shinji
Yoshimatsu, Masayoshi
Kitamura, Morimasa
Suehiro, Atsushi
Hiwatashi, Nao
Yamashita, Masaru
Tateya, Ichiro
Omori, Koichi
author_facet Nakamura, Ryosuke
Katsuno, Tatsuya
Kishimoto, Yo
Kaba, Shinji
Yoshimatsu, Masayoshi
Kitamura, Morimasa
Suehiro, Atsushi
Hiwatashi, Nao
Yamashita, Masaru
Tateya, Ichiro
Omori, Koichi
author_sort Nakamura, Ryosuke
collection PubMed
description Multiciliated epithelial cells in the airway are essential for mucociliary clearance. Their function relies on coordinated, metachronal and directional ciliary beating, appropriate mucus secretion and airway surface hydration. However, current conventional methods for observing human airway ciliary movement require ciliated cells to be detached from airway tissues. Determining the directionality of cilia is difficult. We developed a novel method to stain airway epithelial cilia to observe their movement without releasing ciliated cells. Human tracheae were obtained from patients (n = 13) who underwent laryngectomies to treat malignancies or swallowing disorders. The tracheae were treated with fluorescently labeled wheat germ agglutinin, which interacts with the acidic mucopolysaccharides present on the cilia. Epithelial surfaces were observed using an epi-fluorescence microscope equipped with a water-immersion objective lens and a high-speed camera. Ciliary movement was observable at 125 fps (13/13 samples). Ciliated cells in close proximity mostly exhibited well-coordinated ciliary beats with similar directionalities. These findings indicated that wheat germ agglutinin renders ciliary beats visible, which is valuable for observing human airway ciliary movements in situ.
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spelling pubmed-74681552020-09-03 A novel method for live imaging of human airway cilia using wheat germ agglutinin Nakamura, Ryosuke Katsuno, Tatsuya Kishimoto, Yo Kaba, Shinji Yoshimatsu, Masayoshi Kitamura, Morimasa Suehiro, Atsushi Hiwatashi, Nao Yamashita, Masaru Tateya, Ichiro Omori, Koichi Sci Rep Article Multiciliated epithelial cells in the airway are essential for mucociliary clearance. Their function relies on coordinated, metachronal and directional ciliary beating, appropriate mucus secretion and airway surface hydration. However, current conventional methods for observing human airway ciliary movement require ciliated cells to be detached from airway tissues. Determining the directionality of cilia is difficult. We developed a novel method to stain airway epithelial cilia to observe their movement without releasing ciliated cells. Human tracheae were obtained from patients (n = 13) who underwent laryngectomies to treat malignancies or swallowing disorders. The tracheae were treated with fluorescently labeled wheat germ agglutinin, which interacts with the acidic mucopolysaccharides present on the cilia. Epithelial surfaces were observed using an epi-fluorescence microscope equipped with a water-immersion objective lens and a high-speed camera. Ciliary movement was observable at 125 fps (13/13 samples). Ciliated cells in close proximity mostly exhibited well-coordinated ciliary beats with similar directionalities. These findings indicated that wheat germ agglutinin renders ciliary beats visible, which is valuable for observing human airway ciliary movements in situ. Nature Publishing Group UK 2020-09-02 /pmc/articles/PMC7468155/ /pubmed/32879324 http://dx.doi.org/10.1038/s41598-020-71049-z Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Nakamura, Ryosuke
Katsuno, Tatsuya
Kishimoto, Yo
Kaba, Shinji
Yoshimatsu, Masayoshi
Kitamura, Morimasa
Suehiro, Atsushi
Hiwatashi, Nao
Yamashita, Masaru
Tateya, Ichiro
Omori, Koichi
A novel method for live imaging of human airway cilia using wheat germ agglutinin
title A novel method for live imaging of human airway cilia using wheat germ agglutinin
title_full A novel method for live imaging of human airway cilia using wheat germ agglutinin
title_fullStr A novel method for live imaging of human airway cilia using wheat germ agglutinin
title_full_unstemmed A novel method for live imaging of human airway cilia using wheat germ agglutinin
title_short A novel method for live imaging of human airway cilia using wheat germ agglutinin
title_sort novel method for live imaging of human airway cilia using wheat germ agglutinin
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7468155/
https://www.ncbi.nlm.nih.gov/pubmed/32879324
http://dx.doi.org/10.1038/s41598-020-71049-z
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