Cargando…

Autofluorescence imaging permits label-free cell type assignment and reveals the dynamic formation of airway secretory cell associated antigen passages (SAPs)

The specific functional properties of a tissue are distributed amongst its component cell types. The various cells act coherently, as an ensemble, in order to execute a physiologic response. Modern approaches for identifying and dissecting novel physiologic mechanisms would benefit from an ability t...

Descripción completa

Detalles Bibliográficos
Autores principales: Shah, Viral S, Hou, Jue, Vinarsky, Vladimir, Xu, Jiajie, Surve, Manalee V, Lin, Charles P, Rajagopal, Jayaraj
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10154029/
https://www.ncbi.nlm.nih.gov/pubmed/36994985
http://dx.doi.org/10.7554/eLife.84375
_version_ 1785036046324465664
author Shah, Viral S
Hou, Jue
Vinarsky, Vladimir
Xu, Jiajie
Surve, Manalee V
Lin, Charles P
Rajagopal, Jayaraj
author_facet Shah, Viral S
Hou, Jue
Vinarsky, Vladimir
Xu, Jiajie
Surve, Manalee V
Lin, Charles P
Rajagopal, Jayaraj
author_sort Shah, Viral S
collection PubMed
description The specific functional properties of a tissue are distributed amongst its component cell types. The various cells act coherently, as an ensemble, in order to execute a physiologic response. Modern approaches for identifying and dissecting novel physiologic mechanisms would benefit from an ability to identify specific cell types in live tissues that could then be imaged in real time. Current techniques require the use of fluorescent genetic reporters that are not only cumbersome, but which only allow the study of three or four cell types at a time. We report a non-invasive imaging modality that capitalizes on the endogenous autofluorescence signatures of the metabolic cofactors NAD(P)H and FAD. By marrying morphological characteristics with autofluorescence signatures, all seven of the airway epithelial cell types can be distinguished simultaneously in mouse tracheal explants in real time. Furthermore, we find that this methodology for direct cell type-specific identification avoids pitfalls associated with the use of ostensibly cell type-specific markers that are, in fact, altered by clinically relevant physiologic stimuli. Finally, we utilize this methodology to interrogate real-time physiology and identify dynamic secretory cell associated antigen passages (SAPs) that form in response to cholinergic stimulus. The identical process has been well documented in the intestine where the dynamic formation of SAPs and goblet cell associated antigen passages (GAPs) enable luminal antigen sampling. Airway secretory cells with SAPs are frequently juxtaposed to antigen presenting cells, suggesting that airway SAPs, like their intestinal counterparts, not only sample antigen but convey their cargo for immune cell processing.
format Online
Article
Text
id pubmed-10154029
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher eLife Sciences Publications, Ltd
record_format MEDLINE/PubMed
spelling pubmed-101540292023-05-03 Autofluorescence imaging permits label-free cell type assignment and reveals the dynamic formation of airway secretory cell associated antigen passages (SAPs) Shah, Viral S Hou, Jue Vinarsky, Vladimir Xu, Jiajie Surve, Manalee V Lin, Charles P Rajagopal, Jayaraj eLife Cell Biology The specific functional properties of a tissue are distributed amongst its component cell types. The various cells act coherently, as an ensemble, in order to execute a physiologic response. Modern approaches for identifying and dissecting novel physiologic mechanisms would benefit from an ability to identify specific cell types in live tissues that could then be imaged in real time. Current techniques require the use of fluorescent genetic reporters that are not only cumbersome, but which only allow the study of three or four cell types at a time. We report a non-invasive imaging modality that capitalizes on the endogenous autofluorescence signatures of the metabolic cofactors NAD(P)H and FAD. By marrying morphological characteristics with autofluorescence signatures, all seven of the airway epithelial cell types can be distinguished simultaneously in mouse tracheal explants in real time. Furthermore, we find that this methodology for direct cell type-specific identification avoids pitfalls associated with the use of ostensibly cell type-specific markers that are, in fact, altered by clinically relevant physiologic stimuli. Finally, we utilize this methodology to interrogate real-time physiology and identify dynamic secretory cell associated antigen passages (SAPs) that form in response to cholinergic stimulus. The identical process has been well documented in the intestine where the dynamic formation of SAPs and goblet cell associated antigen passages (GAPs) enable luminal antigen sampling. Airway secretory cells with SAPs are frequently juxtaposed to antigen presenting cells, suggesting that airway SAPs, like their intestinal counterparts, not only sample antigen but convey their cargo for immune cell processing. eLife Sciences Publications, Ltd 2023-03-30 /pmc/articles/PMC10154029/ /pubmed/36994985 http://dx.doi.org/10.7554/eLife.84375 Text en © 2023, Shah, Hou et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Cell Biology
Shah, Viral S
Hou, Jue
Vinarsky, Vladimir
Xu, Jiajie
Surve, Manalee V
Lin, Charles P
Rajagopal, Jayaraj
Autofluorescence imaging permits label-free cell type assignment and reveals the dynamic formation of airway secretory cell associated antigen passages (SAPs)
title Autofluorescence imaging permits label-free cell type assignment and reveals the dynamic formation of airway secretory cell associated antigen passages (SAPs)
title_full Autofluorescence imaging permits label-free cell type assignment and reveals the dynamic formation of airway secretory cell associated antigen passages (SAPs)
title_fullStr Autofluorescence imaging permits label-free cell type assignment and reveals the dynamic formation of airway secretory cell associated antigen passages (SAPs)
title_full_unstemmed Autofluorescence imaging permits label-free cell type assignment and reveals the dynamic formation of airway secretory cell associated antigen passages (SAPs)
title_short Autofluorescence imaging permits label-free cell type assignment and reveals the dynamic formation of airway secretory cell associated antigen passages (SAPs)
title_sort autofluorescence imaging permits label-free cell type assignment and reveals the dynamic formation of airway secretory cell associated antigen passages (saps)
topic Cell Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10154029/
https://www.ncbi.nlm.nih.gov/pubmed/36994985
http://dx.doi.org/10.7554/eLife.84375
work_keys_str_mv AT shahvirals autofluorescenceimagingpermitslabelfreecelltypeassignmentandrevealsthedynamicformationofairwaysecretorycellassociatedantigenpassagessaps
AT houjue autofluorescenceimagingpermitslabelfreecelltypeassignmentandrevealsthedynamicformationofairwaysecretorycellassociatedantigenpassagessaps
AT vinarskyvladimir autofluorescenceimagingpermitslabelfreecelltypeassignmentandrevealsthedynamicformationofairwaysecretorycellassociatedantigenpassagessaps
AT xujiajie autofluorescenceimagingpermitslabelfreecelltypeassignmentandrevealsthedynamicformationofairwaysecretorycellassociatedantigenpassagessaps
AT survemanaleev autofluorescenceimagingpermitslabelfreecelltypeassignmentandrevealsthedynamicformationofairwaysecretorycellassociatedantigenpassagessaps
AT lincharlesp autofluorescenceimagingpermitslabelfreecelltypeassignmentandrevealsthedynamicformationofairwaysecretorycellassociatedantigenpassagessaps
AT rajagopaljayaraj autofluorescenceimagingpermitslabelfreecelltypeassignmentandrevealsthedynamicformationofairwaysecretorycellassociatedantigenpassagessaps