Membrane Capacitance Recordings Resolve Dynamics and Complexity of Receptor-mediated Endocytosis in Wnt Signalling

Overview

Journal Sci Rep

Specialty Science

Date 2019 Sep 12

PMID 31506500

Citations 9

Authors

Vera Bandmann

Ann Schirin Mirsanaye

Johanna Schafer

Gerhard Thiel

Thomas Holstein

Melanie Mikosch-Wersching

Affiliations

Soon will be listed here.

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 different 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 specifically triggered endocytotic activity, which started immediately upon ligand binding and ceased within a period of ten minutes. By using specific 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 findings show that membrane capacitance recordings allow the resolution of complex cellular processes in plasma membrane signalling pathways in great detail.

Citing Articles

Improved Sensitivity in a Modified Berkeley Red Sensor of Transmembrane Potential.

Navarro M, Gerstner N, Lipman S, Dolgonos G, Miller E ACS Chem Biol. 2024; 19(10):2214-2219.

PMID: 39358835 PMC: 11648967. DOI: 10.1021/acschembio.4c00442.

Optical constraints on two-photon voltage imaging.

Phil Brooks 3rd F, Davis H, Wong-Campos J, Cohen A Neurophotonics. 2024; 11(3):035007.

PMID: 39139631 PMC: 11321468. DOI: 10.1117/1.NPh.11.3.035007.

Transient suppression of Wnt signaling in poor-quality buffalo oocytes improves their developmental competence.

Ahuja K, Batra V, Kumar R, Datta T Front Vet Sci. 2024; 10:1324647.

PMID: 38274663 PMC: 10808588. DOI: 10.3389/fvets.2023.1324647.

Mechanisms of aquaporin-4 vesicular trafficking in mammalian cells.

Markou A, Kitchen P, Aldabbagh A, Repici M, Salman M, Bill R J Neurochem. 2023; 168(2):100-114.

PMID: 38102893 PMC: 10953025. DOI: 10.1111/jnc.16029.

Optical constraints on two-photon voltage imaging.

Phil Brooks 3rd F, Davis H, Wong-Campos J, Cohen A bioRxiv. 2023; .

PMID: 38014011 PMC: 10680948. DOI: 10.1101/2023.11.18.567441.

References

Koo B, Spit M, Jordens I, Low T, Stange D, van de Wetering M . Tumour suppressor RNF43 is a stem-cell E3 ligase that induces endocytosis of Wnt receptors. Nature. 2012; 488(7413):665-9. DOI: 10.1038/nature11308. View

Bandmann V, Homann U . Clathrin-independent endocytosis contributes to uptake of glucose into BY-2 protoplasts. Plant J. 2012; 70(4):578-84. DOI: 10.1111/j.1365-313X.2011.04892.x. View

Kikuchi A, Yamamoto H, Sato A . Selective activation mechanisms of Wnt signaling pathways. Trends Cell Biol. 2009; 19(3):119-29. DOI: 10.1016/j.tcb.2009.01.003. View

Hagemann A, Kurz J, Kauffeld S, Chen Q, Reeves P, Weber S . In vivo analysis of formation and endocytosis of the Wnt/β-catenin signaling complex in zebrafish embryos. J Cell Sci. 2014; 127(Pt 18):3970-82. PMC: 4163645. DOI: 10.1242/jcs.148767. View

Mahapatra S, Fan F, Lou X . Tissue-specific dynamin-1 deletion at the calyx of Held decreases short-term depression through a mechanism distinct from vesicle resupply. Proc Natl Acad Sci U S A. 2016; 113(22):E3150-8. PMC: 4896691. DOI: 10.1073/pnas.1520937113. View

Basagiannis D, Zografou S, Galanopoulou K, Christoforidis S . Dynasore impairs VEGFR2 signalling in an endocytosis-independent manner. Sci Rep. 2017; 7:45035. PMC: 5361198. DOI: 10.1038/srep45035. View

Neher E, Marty A . Discrete changes of cell membrane capacitance observed under conditions of enhanced secretion in bovine adrenal chromaffin cells. Proc Natl Acad Sci U S A. 1982; 79(21):6712-6. PMC: 347199. DOI: 10.1073/pnas.79.21.6712. View

Ding V, Ling L, Natarajan S, Yap M, Cool S, Choo A . FGF-2 modulates Wnt signaling in undifferentiated hESC and iPS cells through activated PI3-K/GSK3beta signaling. J Cell Physiol. 2010; 225(2):417-28. DOI: 10.1002/jcp.22214. View

Kirchhausen T, Owen D, Harrison S . Molecular structure, function, and dynamics of clathrin-mediated membrane traffic. Cold Spring Harb Perspect Biol. 2014; 6(5):a016725. PMC: 3996469. DOI: 10.1101/cshperspect.a016725. View

10.

Ohkawara B, Glinka A, Niehrs C . Rspo3 binds syndecan 4 and induces Wnt/PCP signaling via clathrin-mediated endocytosis to promote morphogenesis. Dev Cell. 2011; 20(3):303-14. DOI: 10.1016/j.devcel.2011.01.006. View

11.

Nusse R, Clevers H . Wnt/β-Catenin Signaling, Disease, and Emerging Therapeutic Modalities. Cell. 2017; 169(6):985-999. DOI: 10.1016/j.cell.2017.05.016. View

12.

Gagliardi M, Hernandez A, McGough I, Vincent J . Inhibitors of endocytosis prevent Wnt/Wingless signalling by reducing the level of basal β-catenin/Armadillo. J Cell Sci. 2014; 127(Pt 22):4918-26. PMC: 4231306. DOI: 10.1242/jcs.155424. View

13.

Kim G, Her J, Han J . Ryk cooperates with Frizzled 7 to promote Wnt11-mediated endocytosis and is essential for Xenopus laevis convergent extension movements. J Cell Biol. 2008; 182(6):1073-82. PMC: 2542470. DOI: 10.1083/jcb.200710188. View

14.

Song J, Wang Z, Ewing R . Integrated analysis of the Wnt responsive proteome in human cells reveals diverse and cell-type specific networks. Mol Biosyst. 2013; 10(1):45-53. PMC: 3909504. DOI: 10.1039/c3mb70417c. View

15.

Taelman V, Dobrowolski R, Plouhinec J, Fuentealba L, Vorwald P, Gumper I . Wnt signaling requires sequestration of glycogen synthase kinase 3 inside multivesicular endosomes. Cell. 2010; 143(7):1136-48. PMC: 3022472. DOI: 10.1016/j.cell.2010.11.034. View

16.

Okamoto Y, Ninomiya H, Miwa S, Masaki T . Cholesterol oxidation switches the internalization pathway of endothelin receptor type A from caveolae to clathrin-coated pits in Chinese hamster ovary cells. J Biol Chem. 2000; 275(9):6439-46. DOI: 10.1074/jbc.275.9.6439. View

17.

Vercauteren D, Vandenbroucke R, Jones A, Rejman J, Demeester J, De Smedt S . The use of inhibitors to study endocytic pathways of gene carriers: optimization and pitfalls. Mol Ther. 2009; 18(3):561-9. PMC: 2839427. DOI: 10.1038/mt.2009.281. View

18.

Lindau M, Neher E . Patch-clamp techniques for time-resolved capacitance measurements in single cells. Pflugers Arch. 1988; 411(2):137-46. DOI: 10.1007/BF00582306. View

19.

Dos Santos T, Varela J, Lynch I, Salvati A, Dawson K . Effects of transport inhibitors on the cellular uptake of carboxylated polystyrene nanoparticles in different cell lines. PLoS One. 2011; 6(9):e24438. PMC: 3176276. DOI: 10.1371/journal.pone.0024438. View

20.

Rituper B, Flasker A, Gucek A, Chowdhury H, Zorec R . Cholesterol and regulated exocytosis: a requirement for unitary exocytotic events. Cell Calcium. 2012; 52(3-4):250-8. DOI: 10.1016/j.ceca.2012.05.009. View