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Post-correlation on-lamella cryo-CLEM reveals the membrane architecture of lamellar bodies

Lamellar bodies (LBs) are surfactant-rich organelles in alveolar cells. LBs disassemble into a lipid-protein network that reduces surface tension and facilitates gas exchange in the alveolar cavity. Current knowledge of LB architecture is predominantly based on electron microscopy studies using disr...

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Autores principales: Klein, Steffen, Wimmer, Benedikt H., Winter, Sophie L., Kolovou, Androniki, Laketa, Vibor, Chlanda, Petr
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7846596/
https://www.ncbi.nlm.nih.gov/pubmed/33514845
http://dx.doi.org/10.1038/s42003-020-01567-z
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author Klein, Steffen
Wimmer, Benedikt H.
Winter, Sophie L.
Kolovou, Androniki
Laketa, Vibor
Chlanda, Petr
author_facet Klein, Steffen
Wimmer, Benedikt H.
Winter, Sophie L.
Kolovou, Androniki
Laketa, Vibor
Chlanda, Petr
author_sort Klein, Steffen
collection PubMed
description Lamellar bodies (LBs) are surfactant-rich organelles in alveolar cells. LBs disassemble into a lipid-protein network that reduces surface tension and facilitates gas exchange in the alveolar cavity. Current knowledge of LB architecture is predominantly based on electron microscopy studies using disruptive sample preparation methods. We established and validated a post-correlation on-lamella cryo-correlative light and electron microscopy approach for cryo-FIB milled cells to structurally characterize and validate the identity of LBs in their unperturbed state. Using deconvolution and 3D image registration, we were able to identify fluorescently labeled membrane structures analyzed by cryo-electron tomography. In situ cryo-electron tomography of A549 cells as well as primary Human Small Airway Epithelial Cells revealed that LBs are composed of membrane sheets frequently attached to the limiting membrane through “T”-junctions. We report a so far undescribed outer membrane dome protein complex (OMDP) on the limiting membrane of LBs. Our data suggest that LB biogenesis is driven by parallel membrane sheet import and by the curvature of the limiting membrane to maximize lipid storage capacity.
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spelling pubmed-78465962021-02-08 Post-correlation on-lamella cryo-CLEM reveals the membrane architecture of lamellar bodies Klein, Steffen Wimmer, Benedikt H. Winter, Sophie L. Kolovou, Androniki Laketa, Vibor Chlanda, Petr Commun Biol Article Lamellar bodies (LBs) are surfactant-rich organelles in alveolar cells. LBs disassemble into a lipid-protein network that reduces surface tension and facilitates gas exchange in the alveolar cavity. Current knowledge of LB architecture is predominantly based on electron microscopy studies using disruptive sample preparation methods. We established and validated a post-correlation on-lamella cryo-correlative light and electron microscopy approach for cryo-FIB milled cells to structurally characterize and validate the identity of LBs in their unperturbed state. Using deconvolution and 3D image registration, we were able to identify fluorescently labeled membrane structures analyzed by cryo-electron tomography. In situ cryo-electron tomography of A549 cells as well as primary Human Small Airway Epithelial Cells revealed that LBs are composed of membrane sheets frequently attached to the limiting membrane through “T”-junctions. We report a so far undescribed outer membrane dome protein complex (OMDP) on the limiting membrane of LBs. Our data suggest that LB biogenesis is driven by parallel membrane sheet import and by the curvature of the limiting membrane to maximize lipid storage capacity. Nature Publishing Group UK 2021-01-29 /pmc/articles/PMC7846596/ /pubmed/33514845 http://dx.doi.org/10.1038/s42003-020-01567-z Text en © The Author(s) 2021 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
Klein, Steffen
Wimmer, Benedikt H.
Winter, Sophie L.
Kolovou, Androniki
Laketa, Vibor
Chlanda, Petr
Post-correlation on-lamella cryo-CLEM reveals the membrane architecture of lamellar bodies
title Post-correlation on-lamella cryo-CLEM reveals the membrane architecture of lamellar bodies
title_full Post-correlation on-lamella cryo-CLEM reveals the membrane architecture of lamellar bodies
title_fullStr Post-correlation on-lamella cryo-CLEM reveals the membrane architecture of lamellar bodies
title_full_unstemmed Post-correlation on-lamella cryo-CLEM reveals the membrane architecture of lamellar bodies
title_short Post-correlation on-lamella cryo-CLEM reveals the membrane architecture of lamellar bodies
title_sort post-correlation on-lamella cryo-clem reveals the membrane architecture of lamellar bodies
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7846596/
https://www.ncbi.nlm.nih.gov/pubmed/33514845
http://dx.doi.org/10.1038/s42003-020-01567-z
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