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Membrane capacitance recordings resolve dynamics and complexity of receptor-mediated endocytosis in Wnt signalling

Bandmann, Vera and Mirsanaye, Ann Schirin and Schäfer, Johanna and Thiel, Gerhard and Holstein, Thomas and Mikosch-Wersching, Melanie (2019):
Membrane capacitance recordings resolve dynamics and complexity of receptor-mediated endocytosis in Wnt signalling.
In: Scientific Reports, 9 (1), Springer Nature, ISSN 2045-2322,
DOI: 10.1038/s41598-019-49082-4,
[Article]

Abstract

Receptor-mediated endocytosis is an essential process in signalling pathways for activation of intracellular signalling cascades. One example is the Wnt signalling pathway that seems to depend on endocytosis of the ligand-receptor complex for initiation of Wnt signal transduction. To date, the roles of diferent endocytic pathways in Wnt signalling, molecular players and the kinetics of the process remain unclear. Here, we monitored endocytosis in Wnt3a and Wnt5a-mediated signalling with membrane capacitance recordings of HEK293 cells. Our measurements revealed a swift and substantial increase in the number of endocytic vesicles. Extracellular Wnt ligands specifcally triggered endocytotic activity, which started immediately upon ligand binding and ceased within a period of ten minutes. By using specifc inhibitors, we were able to separate Wnt-induced endocytosis into two independent pathways. We demonstrate that canonical Wnt3a is taken up mainly by clathrin-independent endocytosis whereas noncanonical Wnt5a is exclusively regulated via clathrin-mediated endocytosis. Our fndings show that membrane capacitance recordings allow the resolution of complex cellular processes in plasma membrane signalling pathways in great detail.

Item Type: Article
Erschienen: 2019
Creators: Bandmann, Vera and Mirsanaye, Ann Schirin and Schäfer, Johanna and Thiel, Gerhard and Holstein, Thomas and Mikosch-Wersching, Melanie
Title: Membrane capacitance recordings resolve dynamics and complexity of receptor-mediated endocytosis in Wnt signalling
Language: English
Abstract:

Receptor-mediated endocytosis is an essential process in signalling pathways for activation of intracellular signalling cascades. One example is the Wnt signalling pathway that seems to depend on endocytosis of the ligand-receptor complex for initiation of Wnt signal transduction. To date, the roles of diferent endocytic pathways in Wnt signalling, molecular players and the kinetics of the process remain unclear. Here, we monitored endocytosis in Wnt3a and Wnt5a-mediated signalling with membrane capacitance recordings of HEK293 cells. Our measurements revealed a swift and substantial increase in the number of endocytic vesicles. Extracellular Wnt ligands specifcally triggered endocytotic activity, which started immediately upon ligand binding and ceased within a period of ten minutes. By using specifc inhibitors, we were able to separate Wnt-induced endocytosis into two independent pathways. We demonstrate that canonical Wnt3a is taken up mainly by clathrin-independent endocytosis whereas noncanonical Wnt5a is exclusively regulated via clathrin-mediated endocytosis. Our fndings show that membrane capacitance recordings allow the resolution of complex cellular processes in plasma membrane signalling pathways in great detail.

Journal or Publication Title: Scientific Reports
Journal volume: 9
Number: 1
Publisher: Springer Nature
Divisions: 10 Department of Biology
10 Department of Biology > Plant Membrane Biophysics
Date Deposited: 03 Nov 2019 20:57
DOI: 10.1038/s41598-019-49082-4
Official URL: https://doi.org/10.1038/s41598-019-49082-4
URN: urn:nbn:de:tuda-tuprints-92218
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