title: The PAR complex controls the spatiotemporal dynamics of F-actin and the MTOC in directionally migrating leukocytes.
abstract: Inflammatory cells acquire a polarized phenotype to migrate towards sites of infection or injury. A conserved polarity complex comprising PAR-3, PAR-6 and atypical protein kinase C (aPKC) relays extracellular polarizing cues to control cytoskeletal and signaling networks affecting morphological and functional polarization. However, there is no evidence that myeloid cells use PAR signaling to migrate vectorially in three-dimensional (3D) environments in vivo. Using genetically encoded bioprobes and high-resolution live imaging, we reveal the existence of F-actin oscillations in the trailing edge and constant repositioning of the microtubule organizing center (MTOC) to direct leukocyte migration in wounded medaka fish larvae (Oryzias latipes). Genetic manipulation in live myeloid cells demonstrates that the catalytic activity of aPKC and the regulated interaction with PAR-3 and PAR-6 are required for consistent F-actin oscillations, MTOC perinuclear mobility, aPKC repositioning and wound-directed migration upstream of Rho kinase (also known as ROCK or ROK) activation. We propose that the PAR complex coordinately controls cytoskeletal changes affecting both the generation of traction force and the directionality of leukocyte migration to sites of injury.
authors.affiliation: Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Scientific Institute, 20132 Milan, Italy.
authors.forename: Carolina Lage
authors.initials: CL
authors.lastname: Crespo
authors.affiliation: IFOM Foundation, Institute FIRC of Molecular Oncology, 20139 Milan, Italy.
authors.forename: Claudio
authors.initials: C
authors.lastname: Vernieri
authors.affiliation: Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, 20147 VI, USA.
authors.forename: Philipp J
authors.initials: PJ
authors.lastname: Keller
authors.affiliation: IFOM Foundation, Institute FIRC of Molecular Oncology, 20139 Milan, Italy.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Garrè
authors.affiliation: Department of Medicine, Raymond and Beverly Sackler Foundation Cardiovascular Laboratory, Yale University, New Haven, 06511 CT, USA.
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Bender
authors.affiliation: Center for Organismal Studies Heidelberg, University of Heidelberg, 69120 Heidelberg, Germany.
authors.forename: Joachim
authors.initials: J
authors.lastname: Wittbrodt
authors.affiliation: Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Scientific Institute, 20132 Milan, Italy Vita-Salute San Raffaele University School of Medicine, 20132 Milan, Italy pardi.ruggero@hsr.it.
authors.forename: Ruggero
authors.initials: R
authors.lastname: Pardi
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D030801:Animals, Genetically Modified; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002465:Cell Movement; D016764:Cell Polarity / Q000235:genetics; D002478:Cells, Cultured; D007962:Leukocytes / Q000502:physiology*; D022101:Microtubule-Organizing Center / Q000502:physiology*; D046912:Multiprotein Complexes / Q000235:genetics; D009154:Mutation / Q000235:genetics; D009990:Oryzias; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*; D054460:rho-Associated Kinases / Q000378:metabolism
chemical_list: D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D046912:Multiprotein Complexes; C534334:Pard6gb protein, zebrafish; D029961:Zebrafish Proteins; C486818:pard3ab protein, zebrafish; D054460:rho-Associated Kinases; C115015:PKC-3 protein; D011493:Protein Kinase C
keywords: 3D Migration; Cell polarity; Cytoskeleton; Inflammation; Leukocyte; PAR complex