Evaluation of the force and spatial dynamics of macrophage podosomes by multi-particle tracking

Methods : a Companion to Methods in Enzymology
Amsha ProagRenaud Poincloux

Abstract

Podosomes are submicron adhesive and mechanosensitive structures formed by macrophages, dendritic cells and osteoclasts that are capable of protruding into the extracellular environment. Built of an F-actin core surrounded by an adhesion ring, podosomes assemble in a network interconnected by acto-myosin cables. They have been shown to display spatiotemporal instability as well as protrusion force oscillations. To analyse the entire population of these unstable structures, we have designed an automated multi-particle tracking adapted to both topographical and fluorescence data. Here we describe in detail this approach and report the measurements of individual and collective characteristics of podosome ensembles, providing an integrated picture of their activity from the complementary angles of organisation, dynamics, mobility and mechanics. We believe that this will lead to a comprehensive view of podosome collective behaviour and deepen our knowledge about the significance of mechanosensing mediated by protrusive structures.

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Citations

Sep 1, 2015·Methods : a Companion to Methods in Enzymology·Michael J SiedlikCeleste M Nelson
Feb 21, 2016·Methods : a Companion to Methods in Enzymology·Nicolas BorghiChristophe Lavelle
Feb 29, 2020·Current Opinion in Hematology·Florian AlonsoElisabeth Génot
Mar 11, 2020·Journal of Cell Science·Robert HerzogStefan Linder
Dec 15, 2019·Journal of Cell Science·Koen van den DriesRenaud Poincloux
May 19, 2021·Cellular Signalling·Yasaswi Gayatri Mishra, Bramanandam Manavathi

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