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A novel live cell imaging assay reveals regulation of endosome maturation

Podinovskaia, Maria and Prescianotto-Baschong, Cristina and Buser, Dominik P. and Spang, Anne. (2021) A novel live cell imaging assay reveals regulation of endosome maturation.

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Abstract

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-dependent endosomal recycling 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.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Growth & Development > Biochemistry (Spang)
UniBasel Contributors:Spang, Anne
Item Type:Working Paper
Publisher:Cold Spring Harbor Laboratory
Number of Pages:53
Note:Publication type according to Uni Basel Research Database: Discussion paper / Internet publication
Language:English
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edoc DOI:
Last Modified:02 Jul 2021 06:41
Deposited On:02 Jul 2021 06:41

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