title: Actin polymerisation regulates integrin-mediated adhesion as well as rigidity of neutrophils.
abstract: Activation of adherent neutrophils causes them to convert from selectin-mediated rolling to integrin-mediated immobilisation and migration. Migration is known to depend on formation and redistribution of filamentous (F) actin, but although immobilisation in seconds parallels early cortical actin polymerisation, no link has been proven. We tested the effect of the actin-polymerising agent jasplakinolide (10 microM) on adhesive and mechanical properties of neutrophils. Pretreated cells were able to adhere and roll on immobilised platelets in a flow-based adhesion assay, but whereas untreated rolling cells became immobilised in seconds when chemotactic formyl peptide (fMLP, 0.1 microM) was superfused over them, the cells treated with jasplakinolide continued rolling. Pretreatment with jasplakinolide also blocked de novo expression of integrin CD11b and shape change which otherwise occurred in minutes after treatment with fMLP. Jasplakinolide directly caused actin polymerisation within neutrophils, evidenced by a marked increase in rigidity (resistance to aspiration into a 5 microm micropipette) and increase in association of actin with the Triton-insoluble cytoskeleton. These results indicate that rearrangement of the actin cytoskeleton regulates integrin-mediated adhesion of activated neutrophils, as well as their migration and mechanical properties.
authors.affiliation: Department of Physiology, The Medical School, The University of Birmingham, United Kingdom.
authors.forename: S
authors.initials: S
authors.lastname: Sheikh
authors.forename: W B
authors.initials: WB
authors.lastname: Gratzer
authors.forename: J C
authors.initials: JC
authors.lastname: Pinder
authors.forename: G B
authors.initials: GB
authors.lastname: Nash
mesh_terms: D000199:Actins / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology*; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D003571:Cytochalasin B / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects / Q000502:physiology; D047630:Depsipeptides; D006801:Humans; D016023:Integrins / Q000502:physiology*; D016177:Macrophage-1 Antigen / Q000096:biosynthesis / Q000187:drug effects; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D010456:Peptides, Cyclic / Q000494:pharmacology; D011108:Polymers / Q000378:metabolism
chemical_list: D000199:Actins; D047630:Depsipeptides; D016023:Integrins; D016177:Macrophage-1 Antigen; D010456:Peptides, Cyclic; D011108:Polymers; C057531:jasplakinolide; D003571:Cytochalasin B; D009240:N-Formylmethionine Leucyl-Phenylalanine