BASHY Dye Platform Enables the Fluorescence Bioimaging of Myelin Debris Phagocytosis by Microglia during Demyelination

Multiple sclerosis (MS) is a demyelinating disease of the central nervous system that is characterized by the presence of demyelinated regions with accumulated myelin lipid debris. Importantly, to allow effective remyelination, such debris must be cleared by microglia. Therefore, the study of microg...

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Autores principales:	Maria V. Pinto, Fábio M. F. Santos, Catarina Barros, Ana Rita Ribeiro, Uwe Pischel, Pedro M. P. Gois, Adelaide Fernandes
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	multiple sclerosis demyelination microglia phagocytosis myelin debris BASHY in vivo imaging Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/26e2372176704b5aa39e29b1b4aaad9e
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spelling	oai:doaj.org-article:26e2372176704b5aa39e29b1b4aaad9e2021-11-25T17:12:10ZBASHY Dye Platform Enables the Fluorescence Bioimaging of Myelin Debris Phagocytosis by Microglia during Demyelination10.3390/cells101131632073-4409https://doaj.org/article/26e2372176704b5aa39e29b1b4aaad9e2021-11-01T00:00:00Zhttps://www.mdpi.com/2073-4409/10/11/3163https://doaj.org/toc/2073-4409Multiple sclerosis (MS) is a demyelinating disease of the central nervous system that is characterized by the presence of demyelinated regions with accumulated myelin lipid debris. Importantly, to allow effective remyelination, such debris must be cleared by microglia. Therefore, the study of microglial activity with sensitive tools is of great interest to better monitor the MS clinical course. Using a boronic acid-based (BASHY) fluorophore, specific for nonpolar lipid aggregates, we aimed to address BASHY’s ability to label nonpolar myelin debris and image myelin clearance in the context of demyelination. Demyelinated ex vivo organotypic cultures (OCSCs) and primary microglia cells were immunostained to evaluate BASHY’s co-localization with myelin debris and also to evaluate BASHY’s specificity for phagocytosing cells. Additionally, mice induced with experimental autoimmune encephalomyelitis (EAE) were injected with BASHY and posteriorly analyzed to evaluate BASHY<sup>+</sup> microglia within demyelinated lesions. Indeed, in our in vitro and ex vivo studies, we showed a significant increase in BASHY labeling in demyelinated OCSCs, mostly co-localized with Iba1-expressing amoeboid/phagocytic microglia. Most importantly, BASHY’s presence was also found within demyelinated areas of EAE mice, essentially co-localizing with lesion-associated Iba1<sup>+</sup> cells, evidencing BASHY’s potential for the in vivo bioimaging of myelin clearance and myelin-carrying microglia in regions of active demyelination.Maria V. PintoFábio M. F. SantosCatarina BarrosAna Rita RibeiroUwe PischelPedro M. P. GoisAdelaide FernandesMDPI AGarticlemultiple sclerosisdemyelinationmicroglia phagocytosismyelin debrisBASHYin vivo imagingBiology (General)QH301-705.5ENCells, Vol 10, Iss 3163, p 3163 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	multiple sclerosis demyelination microglia phagocytosis myelin debris BASHY in vivo imaging Biology (General) QH301-705.5
spellingShingle	multiple sclerosis demyelination microglia phagocytosis myelin debris BASHY in vivo imaging Biology (General) QH301-705.5 Maria V. Pinto Fábio M. F. Santos Catarina Barros Ana Rita Ribeiro Uwe Pischel Pedro M. P. Gois Adelaide Fernandes BASHY Dye Platform Enables the Fluorescence Bioimaging of Myelin Debris Phagocytosis by Microglia during Demyelination
description	Multiple sclerosis (MS) is a demyelinating disease of the central nervous system that is characterized by the presence of demyelinated regions with accumulated myelin lipid debris. Importantly, to allow effective remyelination, such debris must be cleared by microglia. Therefore, the study of microglial activity with sensitive tools is of great interest to better monitor the MS clinical course. Using a boronic acid-based (BASHY) fluorophore, specific for nonpolar lipid aggregates, we aimed to address BASHY’s ability to label nonpolar myelin debris and image myelin clearance in the context of demyelination. Demyelinated ex vivo organotypic cultures (OCSCs) and primary microglia cells were immunostained to evaluate BASHY’s co-localization with myelin debris and also to evaluate BASHY’s specificity for phagocytosing cells. Additionally, mice induced with experimental autoimmune encephalomyelitis (EAE) were injected with BASHY and posteriorly analyzed to evaluate BASHY<sup>+</sup> microglia within demyelinated lesions. Indeed, in our in vitro and ex vivo studies, we showed a significant increase in BASHY labeling in demyelinated OCSCs, mostly co-localized with Iba1-expressing amoeboid/phagocytic microglia. Most importantly, BASHY’s presence was also found within demyelinated areas of EAE mice, essentially co-localizing with lesion-associated Iba1<sup>+</sup> cells, evidencing BASHY’s potential for the in vivo bioimaging of myelin clearance and myelin-carrying microglia in regions of active demyelination.
format	article
author	Maria V. Pinto Fábio M. F. Santos Catarina Barros Ana Rita Ribeiro Uwe Pischel Pedro M. P. Gois Adelaide Fernandes
author_facet	Maria V. Pinto Fábio M. F. Santos Catarina Barros Ana Rita Ribeiro Uwe Pischel Pedro M. P. Gois Adelaide Fernandes
author_sort	Maria V. Pinto
title	BASHY Dye Platform Enables the Fluorescence Bioimaging of Myelin Debris Phagocytosis by Microglia during Demyelination
title_short	BASHY Dye Platform Enables the Fluorescence Bioimaging of Myelin Debris Phagocytosis by Microglia during Demyelination
title_full	BASHY Dye Platform Enables the Fluorescence Bioimaging of Myelin Debris Phagocytosis by Microglia during Demyelination
title_fullStr	BASHY Dye Platform Enables the Fluorescence Bioimaging of Myelin Debris Phagocytosis by Microglia during Demyelination
title_full_unstemmed	BASHY Dye Platform Enables the Fluorescence Bioimaging of Myelin Debris Phagocytosis by Microglia during Demyelination
title_sort	bashy dye platform enables the fluorescence bioimaging of myelin debris phagocytosis by microglia during demyelination
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/26e2372176704b5aa39e29b1b4aaad9e
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_version_	1718412636892168192

BASHY Dye Platform Enables the Fluorescence Bioimaging of Myelin Debris Phagocytosis by Microglia during Demyelination

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