Show simple item record

dc.contributor.authorAnderson, Stephen I.
dc.contributor.authorBehrendt, Barbara
dc.contributor.authorMachesky, Laura M.
dc.contributor.authorInsall, Robert H.
dc.contributor.authorNash, Gerard B.
dc.date.accessioned2008-03-18T11:21:55Z
dc.date.available2008-03-18T11:21:55Z
dc.date.issued2003
dc.identifier.citationCell Motility and the Cytoskeleton, 54(2): 135-146
dc.identifier.issn0886-1544
dc.identifier.pmid12529859
dc.identifier.doi10.1002/cm.10091
dc.identifier.urihttp://hdl.handle.net/2436/20981
dc.description.abstractNeutrophils migrate rapidly by co-ordinating regulation of their beta2-integrin adhesion with turnover of filamentous F-actin. The seven-protein Arp2/3 complex regulates actin polymerisation upon activation by proteins of the WASP-family. To investigate links between actin polymerisation, adhesion, and migration, we used a novel osmotic-shock method to load neutrophils with peptides: (1). WASP-WA and Scar-WA (which incorporate the actin- and Arp2/3-binding regions of WASP and Scar1), to compete with endogenous WASP-family members; (2). proline rich motifs (PRM) from the ActA protein of L. monocytogenes or from vinculin, which bind vasodilator-stimulated phosphoprotein (VASP), a regulator of cytoskeleton assembly. In a flow system, rolling-adherent neutrophils were stimulated with formyl tri-peptide. This caused rapid immobilisation, followed by migration with increasing velocity, supported by activated beta2-integrin CD11b/CD18. Loading ActA PRM (but not vinculin PRM) caused concentration-dependent reduction in migration velocity. At the highest concentration, unstimulated neutrophils had elevated F-actin and were rigid, but could not change their F-actin content or shape upon stimulation. Scar-WA also caused marked reduction in migration rate, but WASP-WA had a lesser effect. Scar-WA did not modify activation-dependent formation of F-actin or change in shape. However, a reduction in rate of downregulation of integrin adhesion appeared to contribute to impaired migration. These studies show that interference in cytoskeletal reorganisation that follows activation in neutrophils, can impair regulation of integrin function as well as motility. They also suggest a role of the Arp2/3 complex and WASP-family in co-ordinating actin polymerisation and integrin function in migrating neutrophils.
dc.language.isoen
dc.publisherWiley Interscience
dc.relation.urlhttp://www3.interscience.wiley.com/journal/102524780/abstract
dc.subjectLeukocytes
dc.subjectLeukocyte migration
dc.subjectCell Adhesion
dc.subjectActin polymerisation
dc.subjectPeptide loading
dc.subjectWASP
dc.subjectArp2/3
dc.subject.meshActin-Related Protein 2
dc.subject.meshActin-Related Protein 3
dc.subject.meshActins
dc.subject.meshAntigens, CD11b
dc.subject.meshAntigens, CD18
dc.subject.meshBacterial Proteins
dc.subject.meshCell Adhesion
dc.subject.meshCell Adhesion Molecules
dc.subject.meshCell Movement
dc.subject.meshCell Size
dc.subject.meshCytoskeletal Proteins
dc.subject.meshCytoskeleton
dc.subject.meshHumans
dc.subject.meshMembrane Proteins
dc.subject.meshMicrofilament Proteins
dc.subject.meshN-Formylmethionine Leucyl-Phenylalanine
dc.subject.meshNeutrophils
dc.subject.meshOsmotic Pressure
dc.subject.meshPeptide Fragments
dc.subject.meshPhosphoproteins
dc.subject.meshPolymers
dc.subject.meshProteins
dc.subject.meshVinculin
dc.subject.meshWiskott-Aldrich Syndrome Protein
dc.subject.meshWiskott-Aldrich Syndrome Protein Family
dc.titleLinked regulation of motility and integrin function in activated migrating neutrophils revealed by interference in remodelling of the cytoskeleton.
dc.typeJournal article
dc.identifier.journalCell Motility and the Cytoskeleton
html.description.abstractNeutrophils migrate rapidly by co-ordinating regulation of their beta2-integrin adhesion with turnover of filamentous F-actin. The seven-protein Arp2/3 complex regulates actin polymerisation upon activation by proteins of the WASP-family. To investigate links between actin polymerisation, adhesion, and migration, we used a novel osmotic-shock method to load neutrophils with peptides: (1). WASP-WA and Scar-WA (which incorporate the actin- and Arp2/3-binding regions of WASP and Scar1), to compete with endogenous WASP-family members; (2). proline rich motifs (PRM) from the ActA protein of L. monocytogenes or from vinculin, which bind vasodilator-stimulated phosphoprotein (VASP), a regulator of cytoskeleton assembly. In a flow system, rolling-adherent neutrophils were stimulated with formyl tri-peptide. This caused rapid immobilisation, followed by migration with increasing velocity, supported by activated beta2-integrin CD11b/CD18. Loading ActA PRM (but not vinculin PRM) caused concentration-dependent reduction in migration velocity. At the highest concentration, unstimulated neutrophils had elevated F-actin and were rigid, but could not change their F-actin content or shape upon stimulation. Scar-WA also caused marked reduction in migration rate, but WASP-WA had a lesser effect. Scar-WA did not modify activation-dependent formation of F-actin or change in shape. However, a reduction in rate of downregulation of integrin adhesion appeared to contribute to impaired migration. These studies show that interference in cytoskeletal reorganisation that follows activation in neutrophils, can impair regulation of integrin function as well as motility. They also suggest a role of the Arp2/3 complex and WASP-family in co-ordinating actin polymerisation and integrin function in migrating neutrophils.


This item appears in the following Collection(s)

Show simple item record