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Scanning electron microscopy of human islet cilia

Human islet primary cilia are vital glucose-regulating organelles whose structure remains uncharacterized. Scanning electron microscopy (SEM) is a useful technique for studying the surface morphology of membrane projections like cilia, but conventional sample preparation does not reveal the submembr...

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Autores principales: Polino, Alexander J., Sviben, Sanja, Melena, Isabella, Piston, David W., Hughes, Jing W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10235940/
https://www.ncbi.nlm.nih.gov/pubmed/37205712
http://dx.doi.org/10.1073/pnas.2302624120
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author Polino, Alexander J.
Sviben, Sanja
Melena, Isabella
Piston, David W.
Hughes, Jing W.
author_facet Polino, Alexander J.
Sviben, Sanja
Melena, Isabella
Piston, David W.
Hughes, Jing W.
author_sort Polino, Alexander J.
collection PubMed
description Human islet primary cilia are vital glucose-regulating organelles whose structure remains uncharacterized. Scanning electron microscopy (SEM) is a useful technique for studying the surface morphology of membrane projections like cilia, but conventional sample preparation does not reveal the submembrane axonemal structure, which holds key implications for ciliary function. To overcome this challenge, we combined SEM with membrane-extraction techniques to examine primary cilia in native human islets. Our data show well-preserved cilia subdomains which demonstrate both expected and unexpected ultrastructural motifs. Morphometric features were quantified when possible, including axonemal length and diameter, microtubule conformations, and chirality. We further describe a ciliary ring, a structure that may be a specialization in human islets. Key findings are correlated with fluorescence microscopy and interpreted in the context of cilia function as a cellular sensor and communications locus in pancreatic islets.
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spelling pubmed-102359402023-06-03 Scanning electron microscopy of human islet cilia Polino, Alexander J. Sviben, Sanja Melena, Isabella Piston, David W. Hughes, Jing W. Proc Natl Acad Sci U S A Biological Sciences Human islet primary cilia are vital glucose-regulating organelles whose structure remains uncharacterized. Scanning electron microscopy (SEM) is a useful technique for studying the surface morphology of membrane projections like cilia, but conventional sample preparation does not reveal the submembrane axonemal structure, which holds key implications for ciliary function. To overcome this challenge, we combined SEM with membrane-extraction techniques to examine primary cilia in native human islets. Our data show well-preserved cilia subdomains which demonstrate both expected and unexpected ultrastructural motifs. Morphometric features were quantified when possible, including axonemal length and diameter, microtubule conformations, and chirality. We further describe a ciliary ring, a structure that may be a specialization in human islets. Key findings are correlated with fluorescence microscopy and interpreted in the context of cilia function as a cellular sensor and communications locus in pancreatic islets. National Academy of Sciences 2023-05-19 2023-05-30 /pmc/articles/PMC10235940/ /pubmed/37205712 http://dx.doi.org/10.1073/pnas.2302624120 Text en Copyright © 2023 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle Biological Sciences
Polino, Alexander J.
Sviben, Sanja
Melena, Isabella
Piston, David W.
Hughes, Jing W.
Scanning electron microscopy of human islet cilia
title Scanning electron microscopy of human islet cilia
title_full Scanning electron microscopy of human islet cilia
title_fullStr Scanning electron microscopy of human islet cilia
title_full_unstemmed Scanning electron microscopy of human islet cilia
title_short Scanning electron microscopy of human islet cilia
title_sort scanning electron microscopy of human islet cilia
topic Biological Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10235940/
https://www.ncbi.nlm.nih.gov/pubmed/37205712
http://dx.doi.org/10.1073/pnas.2302624120
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