title: Filopodia act as phagocytic tentacles and pull with discrete steps and a load-dependent velocity.
abstract: Filopodia are thin, spike-like cell surface protrusions containing bundles of parallel actin filaments. So far, filopodial dynamics has mainly been studied in the context of cell motility on coverslip-adherent filopodia by using fluorescence and differential interference contrast (DIC) microscopy. In this study, we used an optical trap and interferometric particle tracking with nanometer precision to measure the three-dimensional dynamics of macrophage filopodia, which were not attached to flat surfaces. We found that filopodia act as cellular tentacles: a few seconds after binding to a particle, filopodia retract and pull the bound particle toward the cell. We observed F-actin-dependent stepwise retraction of filopodia with a mean step size of 36 nm, suggesting molecular motor activity during filopodial pulling. Remarkably, this intracellular stepping motion, which was measured at counteracting forces of up to 19 pN, was transmitted to the extracellular tracked particle via the filopodial F-actin bundle and the cell membrane. The pulling velocity depended strongly on the counteracting force and ranged between 600 nm/s at forces <1 pN and approximately 40 nm/s at forces >15 pN. This result provides an explanation of the significant differences in filopodial retraction velocities previously reported in the literature. The measured filopodial retraction force-velocity relationship is in agreement with a model for force-dependent multiple motor kinetics.
authors.affiliation: European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany. holger.kress@yale.edu
authors.forename: Holger
authors.initials: H
authors.lastname: Kress
authors.forename: Ernst H K
authors.initials: EH
authors.lastname: Stelzer
authors.forename: Daniela
authors.initials: D
authors.lastname: Holzer
authors.forename: Folma
authors.initials: F
authors.lastname: Buss
authors.forename: Gareth
authors.initials: G
authors.lastname: Griffiths
authors.forename: Alexander
authors.initials: A
authors.lastname: Rohrbach
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000502:physiology; D002460:Cell Line; D008264:Macrophages / Q000502:physiology; D051379:Mice; D008863:Microspheres; D008954:Models, Biological; D010587:Phagocytosis / Q000502:physiology*; D011554:Pseudopodia / Q000502:physiology*