A novel live-cell imaging assay reveals regulation of endosome maturation

Cell-cell communication is an essential process in life, with endosomes acting as key organelles for regulating uptake and secretion of signaling molecules. Endocytosed material is accepted by the sorting endosome where it either is sorted for recycling or remains in the endosome as it matures to be...

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Autores principales:	Maria Podinovskaia, Cristina Prescianotto-Baschong, Dominik P Buser, Anne Spang
Formato:	article
Lenguaje:	EN
Publicado:	eLife Sciences Publications Ltd 2021
Materias:	endocytosis endosome maturation Rab GTPases live cell imaging assay organelle acidification membrane transport Medicine R Science Q Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/4b601ec39ade4cea9b69453c24f22015
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id	oai:doaj.org-article:4b601ec39ade4cea9b69453c24f22015
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spelling	oai:doaj.org-article:4b601ec39ade4cea9b69453c24f220152021-12-01T08:10:49ZA novel live-cell imaging assay reveals regulation of endosome maturation10.7554/eLife.709822050-084Xe70982https://doaj.org/article/4b601ec39ade4cea9b69453c24f220152021-11-01T00:00:00Zhttps://elifesciences.org/articles/70982https://doaj.org/toc/2050-084XCell-cell communication is an essential process in life, with endosomes acting as key organelles for regulating uptake and secretion of signaling molecules. Endocytosed material is accepted by the sorting endosome where it either is sorted for recycling or remains in the endosome as it matures to be degraded in the lysosome. Investigation of the endosome maturation process has been hampered by the small size and rapid movement of endosomes in most cellular systems. Here, we report an easy versatile live-cell imaging assay to monitor endosome maturation kinetics, which can be applied to a variety of mammalian cell types. Acute ionophore treatment led to enlarged early endosomal compartments that matured into late endosomes and fused with lysosomes to form endolysosomes. Rab5-to-Rab7 conversion and PI(3)P formation and turn over were recapitulated with this assay and could be observed with a standard widefield microscope. We used this approach to show that Snx1 and Rab11-positive recycling endosome recruitment occurred throughout endosome maturation and was uncoupled from Rab conversion. In contrast, efficient endosomal acidification was dependent on Rab conversion. The assay provides a powerful tool to further unravel various aspects of endosome maturation.Maria PodinovskaiaCristina Prescianotto-BaschongDominik P BuserAnne SpangeLife Sciences Publications Ltdarticleendocytosisendosome maturationRab GTPaseslive cell imaging assayorganelle acidificationmembrane transportMedicineRScienceQBiology (General)QH301-705.5ENeLife, Vol 10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	endocytosis endosome maturation Rab GTPases live cell imaging assay organelle acidification membrane transport Medicine R Science Q Biology (General) QH301-705.5
spellingShingle	endocytosis endosome maturation Rab GTPases live cell imaging assay organelle acidification membrane transport Medicine R Science Q Biology (General) QH301-705.5 Maria Podinovskaia Cristina Prescianotto-Baschong Dominik P Buser Anne Spang A novel live-cell imaging assay reveals regulation of endosome maturation
description	Cell-cell communication is an essential process in life, with endosomes acting as key organelles for regulating uptake and secretion of signaling molecules. Endocytosed material is accepted by the sorting endosome where it either is sorted for recycling or remains in the endosome as it matures to be degraded in the lysosome. Investigation of the endosome maturation process has been hampered by the small size and rapid movement of endosomes in most cellular systems. Here, we report an easy versatile live-cell imaging assay to monitor endosome maturation kinetics, which can be applied to a variety of mammalian cell types. Acute ionophore treatment led to enlarged early endosomal compartments that matured into late endosomes and fused with lysosomes to form endolysosomes. Rab5-to-Rab7 conversion and PI(3)P formation and turn over were recapitulated with this assay and could be observed with a standard widefield microscope. We used this approach to show that Snx1 and Rab11-positive recycling endosome recruitment occurred throughout endosome maturation and was uncoupled from Rab conversion. In contrast, efficient endosomal acidification was dependent on Rab conversion. The assay provides a powerful tool to further unravel various aspects of endosome maturation.
format	article
author	Maria Podinovskaia Cristina Prescianotto-Baschong Dominik P Buser Anne Spang
author_facet	Maria Podinovskaia Cristina Prescianotto-Baschong Dominik P Buser Anne Spang
author_sort	Maria Podinovskaia
title	A novel live-cell imaging assay reveals regulation of endosome maturation
title_short	A novel live-cell imaging assay reveals regulation of endosome maturation
title_full	A novel live-cell imaging assay reveals regulation of endosome maturation
title_fullStr	A novel live-cell imaging assay reveals regulation of endosome maturation
title_full_unstemmed	A novel live-cell imaging assay reveals regulation of endosome maturation
title_sort	novel live-cell imaging assay reveals regulation of endosome maturation
publisher	eLife Sciences Publications Ltd
publishDate	2021
url	https://doaj.org/article/4b601ec39ade4cea9b69453c24f22015
work_keys_str_mv	AT mariapodinovskaia anovellivecellimagingassayrevealsregulationofendosomematuration AT cristinaprescianottobaschong anovellivecellimagingassayrevealsregulationofendosomematuration AT dominikpbuser anovellivecellimagingassayrevealsregulationofendosomematuration AT annespang anovellivecellimagingassayrevealsregulationofendosomematuration AT mariapodinovskaia novellivecellimagingassayrevealsregulationofendosomematuration AT cristinaprescianottobaschong novellivecellimagingassayrevealsregulationofendosomematuration AT dominikpbuser novellivecellimagingassayrevealsregulationofendosomematuration AT annespang novellivecellimagingassayrevealsregulationofendosomematuration
_version_	1718405410505883648

A novel live-cell imaging assay reveals regulation of endosome maturation

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