New imaging tools reveal live cellular collagen secretion, fibril dynamics and network organisation

Abstract Although light microscopy has been used to examine the early trafficking of collagen within the cell, much of our understanding of the detailed organisation of cell deposited collagen is from static electron microscopy studies. To understand the dynamics of live cell collagen deposition and...

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Main Authors: Olivia Kent, Eleanor R. Casey, Max Brown, Steven Bell, Matthew C. Ehrman, Michael J. Flagler, Arto Määttä, Adam M. Benham, Timothy J. Hawkins
Format: Article
Language:English
Published: Nature Portfolio 2025-04-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-025-96280-4
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author Olivia Kent
Eleanor R. Casey
Max Brown
Steven Bell
Matthew C. Ehrman
Michael J. Flagler
Arto Määttä
Adam M. Benham
Timothy J. Hawkins
author_facet Olivia Kent
Eleanor R. Casey
Max Brown
Steven Bell
Matthew C. Ehrman
Michael J. Flagler
Arto Määttä
Adam M. Benham
Timothy J. Hawkins
author_sort Olivia Kent
collection DOAJ
description Abstract Although light microscopy has been used to examine the early trafficking of collagen within the cell, much of our understanding of the detailed organisation of cell deposited collagen is from static electron microscopy studies. To understand the dynamics of live cell collagen deposition and fibril organisation, we generated a bright photostable mNGCol1α2 fusion protein and employed a range of microscopy techniques to follow its intracellular transport and elucidate extracellular fibril formation. Our findings reveal the dynamics of fibril growth and the dynamic nature of collagen network interactions at the cellular level. Notably we observed molecular events that build network organisation, including fibril bundling, bifurcation, directionality along existing fibrils, and looping/intertwining behaviours. Strikingly, mNGCol1α2 fluorescence intensity maxima can mark a fibril before another growing collagen fibril intersects at this location. Real-time, high-resolution imaging of collagen has enabled fibrillogenesis and organisational dynamics to be visualised together in an actively secreting cellular system. We also show that the N-terminal protease site is not an absolute requirement for collagen fibril incorporation. This approach paves the way for assessing the dynamic organisation and assembly of collagen into the extracellular matrix in skin models and other tissues during health, ageing and disease.
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spelling doaj-art-0b2ceea0ea874147845970661659fb8b2025-08-20T03:13:57ZengNature PortfolioScientific Reports2045-23222025-04-0115111510.1038/s41598-025-96280-4New imaging tools reveal live cellular collagen secretion, fibril dynamics and network organisationOlivia Kent0Eleanor R. Casey1Max Brown2Steven Bell3Matthew C. Ehrman4Michael J. Flagler5Arto Määttä6Adam M. Benham7Timothy J. Hawkins8Department of Biosciences, Durham UniversityDepartment of Biosciences, Durham UniversityThe Procter & Gamble Company, Newcastle Innovation CentreDepartment of Biosciences, Durham UniversityProcter & Gamble International Operations SA SG BranchThe Procter & Gamble CompanyDepartment of Biosciences, Durham UniversityDepartment of Biosciences, Durham UniversityDepartment of Biosciences, Durham UniversityAbstract Although light microscopy has been used to examine the early trafficking of collagen within the cell, much of our understanding of the detailed organisation of cell deposited collagen is from static electron microscopy studies. To understand the dynamics of live cell collagen deposition and fibril organisation, we generated a bright photostable mNGCol1α2 fusion protein and employed a range of microscopy techniques to follow its intracellular transport and elucidate extracellular fibril formation. Our findings reveal the dynamics of fibril growth and the dynamic nature of collagen network interactions at the cellular level. Notably we observed molecular events that build network organisation, including fibril bundling, bifurcation, directionality along existing fibrils, and looping/intertwining behaviours. Strikingly, mNGCol1α2 fluorescence intensity maxima can mark a fibril before another growing collagen fibril intersects at this location. Real-time, high-resolution imaging of collagen has enabled fibrillogenesis and organisational dynamics to be visualised together in an actively secreting cellular system. We also show that the N-terminal protease site is not an absolute requirement for collagen fibril incorporation. This approach paves the way for assessing the dynamic organisation and assembly of collagen into the extracellular matrix in skin models and other tissues during health, ageing and disease.https://doi.org/10.1038/s41598-025-96280-4
spellingShingle Olivia Kent
Eleanor R. Casey
Max Brown
Steven Bell
Matthew C. Ehrman
Michael J. Flagler
Arto Määttä
Adam M. Benham
Timothy J. Hawkins
New imaging tools reveal live cellular collagen secretion, fibril dynamics and network organisation
Scientific Reports
title New imaging tools reveal live cellular collagen secretion, fibril dynamics and network organisation
title_full New imaging tools reveal live cellular collagen secretion, fibril dynamics and network organisation
title_fullStr New imaging tools reveal live cellular collagen secretion, fibril dynamics and network organisation
title_full_unstemmed New imaging tools reveal live cellular collagen secretion, fibril dynamics and network organisation
title_short New imaging tools reveal live cellular collagen secretion, fibril dynamics and network organisation
title_sort new imaging tools reveal live cellular collagen secretion fibril dynamics and network organisation
url https://doi.org/10.1038/s41598-025-96280-4
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