title: F-actin waves, actin cortex disassembly and focal exocytosis driven by actin-phosphoinositide positive feedback.
abstract: Actin polymerization is controlled by the phosphoinositide composition of the plasma membrane. However, the molecular mechanisms underlying the spatiotemporal regulation of actin network organization over extended length scales are still unclear. To observe phosphoinositide-dependent cytoskeletal dynamics we combined the model system of frustrated phagocytosis, total internal reflection microscopy and manipulation of the buffer tonicity. We found that macrophages interacting with IgG-coated glass substrates formed circular F-actin waves on their ventral surface enclosing a region of plasma membrane devoid of cortical actin. Plasma membrane free of actin cortex was strongly depleted of PI(4,5)P2 , but enriched in PI(3,4)P2 and displayed a fivefold increase in exocytosis. Wave formation could be promoted by application of a hypotonic shock. The actin waves were characteristic of a bistable wavefront at the boundary between the regions of membrane containing and lacking cortical actin. Phosphoinositide modifiers and RhoGTPase activities dramatically redistributed with respect to the wavefronts, which often exhibited spatial oscillations. Perturbation of either lipid or actin cytoskeleton-related pathways led to rapid loss of both the polarized lipid distribution and the wavefront. As waves travelled over the plasma membrane, wavefront actin was seen to rapidly polymerize and depolymerize at pre-existing clusters of FcγRIIA, coincident with rapid changes in lipid composition. Thus the potential of receptors to support rapid F-actin polymerization appears to depend acutely on the local concentrations of multiple lipid species. We propose that interdependence through positive feedback from the cytoskeleton to lipid modifiers leads to coordinated local cortex remodeling, focal exocytosis, and organizes extended actin networks.
authors.affiliation: Mechanobiology Institute, National University of Singapore, 5A Engineering Drive 1, Singapore, 117411, Singapore.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Masters
authors.affiliation: Mechanobiology Institute, National University of Singapore, 5A Engineering Drive 1, Singapore, 117411, Singapore.|Department of Biological Sciences, Columbia University, New York, New York, 10027.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Sheetz
authors.affiliation: Mechanobiology Institute, National University of Singapore, 5A Engineering Drive 1, Singapore, 117411, Singapore.
authors.forename: Nils C
authors.initials: NC
authors.lastname: Gauthier
mesh_terms: D000199:Actins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D005089:Exocytosis; D010587:Phagocytosis; D010716:Phosphatidylinositols / Q000378:metabolism*
chemical_list: D000199:Actins; D010716:Phosphatidylinositols
keywords: F-actin waves; bistability; exocytosis; phagocytosis; phosphoinositide