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Membrane tension spatially organizes lysosomal exocytosis

View ORCID ProfileHugo Lachuer, View ORCID ProfileLaurent Le, View ORCID ProfileSandrine Lévêque-Fort, View ORCID ProfileBruno Goud, View ORCID ProfileKristine Schauer
doi: https://doi.org/10.1101/2022.04.22.489160
Hugo Lachuer
1Cell Biology and Cancer Unit, Institut Curie, PSL Research University, Sorbonne Université, CNRS UMR144, 75005 Paris, France
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  • ORCID record for Hugo Lachuer
Laurent Le
2Université Paris-Saclay, CNRS, Institut des Sciences Moléculaires d’Orsay, 91405, Orsay, France
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Sandrine Lévêque-Fort
2Université Paris-Saclay, CNRS, Institut des Sciences Moléculaires d’Orsay, 91405, Orsay, France
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Bruno Goud
1Cell Biology and Cancer Unit, Institut Curie, PSL Research University, Sorbonne Université, CNRS UMR144, 75005 Paris, France
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Kristine Schauer
1Cell Biology and Cancer Unit, Institut Curie, PSL Research University, Sorbonne Université, CNRS UMR144, 75005 Paris, France
3Tumor Cell Dynamics Unit, Inserm U1279, Gustave Roussy Institute, Université Paris-Saclay, 94800 Villejuif, France
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  • For correspondence: kristine.schauer@gustaveroussy.fr
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Abstract

Lysosomal exocytosis is involved in many key cellular processes but its spatio-temporal regulation is poorly known. Using total internal reflection fluorescence microscopy (TIRFM) and spatial statistics, we observed that lysosomal exocytosis is not random at the adhesive part of the plasma membrane of RPE1 cells but clustered at different scales. Although the rate of exocytosis is regulated by the actin cytoskeleton, neither interfering with actin or microtubule dynamics by drug treatments alters its spatial organization. Exocytosis events partially co-appear at focal adhesions (FAs) and their clustering is reduced upon removal of FAs. Changes in membrane tension following a hypo-osmotic shock or treatment with methyl-β-cyclodextrin was found to increase clustering. To investigate the link between FAs and membrane tension, cells were cultured on adhesive ring-shaped micropatterns, which allows to control the spatial organization of FAs. By using a combination of TIRFM and fluorescence lifetime imaging microscopy (FLIM) microscopy, we revealed the existence of a radial gradient in membrane tension. By changing the diameter of micropatterned substrates, we further showed that this gradient as well as the extent of exocytosis clustering can be controlled. Together, our data indicate that the spatial clustering of lysosomal exocytosis relies on membrane tension patterning controlled by the spatial organization of FAs.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted April 22, 2022.
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Membrane tension spatially organizes lysosomal exocytosis
Hugo Lachuer, Laurent Le, Sandrine Lévêque-Fort, Bruno Goud, Kristine Schauer
bioRxiv 2022.04.22.489160; doi: https://doi.org/10.1101/2022.04.22.489160
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Membrane tension spatially organizes lysosomal exocytosis
Hugo Lachuer, Laurent Le, Sandrine Lévêque-Fort, Bruno Goud, Kristine Schauer
bioRxiv 2022.04.22.489160; doi: https://doi.org/10.1101/2022.04.22.489160

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