Measuring forces at the leading edge: a force assay for cell motility

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dc.citation.firstpage204en_US
dc.citation.issueNumber1en_US
dc.citation.journalTitleIntegrative Biologyen_US
dc.citation.lastpage214en_US
dc.citation.volumeNumber5en_US
dc.contributor.authorFarrell, Brendaen_US
dc.contributor.authorQian, Fengen_US
dc.contributor.authorKolomeisky, Anatolyen_US
dc.contributor.authorAnvari, Bahmanen_US
dc.contributor.authorBrownell, William E.en_US
dc.date.accessioned2017-07-31T18:12:32Zen_US
dc.date.available2017-07-31T18:12:32Zen_US
dc.date.issued2013en_US
dc.description.abstractCancer cells become mobile by remodelling their cytoskeleton to form migratory structures. This transformation is dominated by actin assembly and disassembly (polymerisation and depolymerisation) in the cytoplasm. Synthesis of filamentous actin produces a force at the leading edge that pushes the plasma membrane forward. We describe an assay to measure the restoring force of the membrane in response to forces generated within the cytoplasm adjacent to the membrane. A laser trap is used to form a long membrane nanotube from a living cell and to measure the axial membrane force at the end of the tube. When the tube, resembling a filopodium, is formed and in a relaxed state the axial membrane force exhibits a positive stationary value. This value reflects the influence of the cytoskeleton that acts to pull the tube back to the cell. A dynamic sawtooth force that rides upon the stationary value is also observed. This force is sensitive to a toxin that affects actin assembly and disassembly, but not affected by agents that influence microtubules and myosin light chain kinase. We deduce from the magnitude and characteristics of dynamic force measurements that it originates from depolymerisation and polymerisation of F-actin. The on- and off-rates, the number of working filaments, and the force per filament (2.5 pN) are determined. We suggest the force-dependent transitions are thermodynamically uncoupled as both the on- and off-rates decrease exponentially with a compressive load. We propose kinetic schemes that require attachment of actin filaments to the membrane during depolymerisation. This demonstrates that actin kinetics can be monitored in a living cell by measuring force at the membrane, and used to probe the mobility of cells including cancer cells.en_US
dc.identifier.citationFarrell, Brenda, Qian, Feng, Kolomeisky, Anatoly, et al.. "Measuring forces at the leading edge: a force assay for cell motility." <i>Integrative Biology,</i> 5, no. 1 (2013) Royal Society of Chemistry: 204-214. http://dx.doi.org/10.1039/C2IB20097J.en_US
dc.identifier.digitalMeasuring_forces_at_the_leading_edgeen_US
dc.identifier.doihttp://dx.doi.org/10.1039/C2IB20097Jen_US
dc.identifier.urihttps://hdl.handle.net/1911/95628en_US
dc.language.isoengen_US
dc.publisherRoyal Society of Chemistryen_US
dc.rightsThis is an author's peer-reviewed final manuscript, as accepted by the publisher. The published article is copyrighted by the Royal Society of Chemistry.en_US
dc.titleMeasuring forces at the leading edge: a force assay for cell motilityen_US
dc.typeJournal articleen_US
dc.type.dcmiTexten_US
dc.type.publicationpost-printen_US
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