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Quantitative single molecule analysis of podoplanin clustering in fibroblastic reticular cells uncovers CD44 function

Lim, Shu En; Joseph, Megan D; de Winde, Charlotte M; Acton, Sophie E; Simoncelli, Sabrina; (2023) Quantitative single molecule analysis of podoplanin clustering in fibroblastic reticular cells uncovers CD44 function. Open Biology , 13 (5) , Article 220377. 10.1098/rsob.220377. Green open access

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Abstract

Upon initial immune challenge, dendritic cells (DCs) migrate to lymph nodes and interact with fibroblastic reticular cells (FRCs) via C-type lectin-like receptor 2 (CLEC-2). CLEC-2 binds to the membrane glycoprotein podoplanin (PDPN) on FRCs, inhibiting actomyosin contractility through the FRC network and permitting lymph node expansion. The hyaluronic acid receptor CD44 is known to be required for FRCs to respond to DCs but the mechanism of action is not fully elucidated. Here, we use DNA-PAINT, a quantitative single molecule super-resolution technique, to visualize and quantify how PDPN clustering is regulated in the plasma membrane of FRCs. Our results indicate that CLEC-2 interaction leads to the formation of large PDPN clusters (i.e. more than 12 proteins per cluster) in a CD44-dependent manner. These results suggest that CD44 expression is required to stabilize large pools of PDPN at the membrane of FRCs upon CLEC-2 interaction, revealing the molecular mechanism through which CD44 facilitates cellular crosstalk between FRCs and DCs.

Type: Article
Title: Quantitative single molecule analysis of podoplanin clustering in fibroblastic reticular cells uncovers CD44 function
Location: England
Open access status: An open access version is available from UCL Discovery
DOI: 10.1098/rsob.220377
Publisher version: https://doi.org/10.1098/rsob.220377
Language: English
Additional information: © 2023 The Authors. Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited.
Keywords: CD44, CLEC-2, DNA-PAINT, podoplanin, qPAINT, super-resolution
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences > London Centre for Nanotechnology
URI: https://discovery.ucl.ac.uk/id/eprint/10169742
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