title: Live cell imaging of membrane/cytoskeleton interactions and membrane topology.
abstract: We elucidate the interaction between actin and specific membrane components, using real time live cell imaging, by delivering probes that enable access to components, that cannot be accessed genetically. We initially investigated the close interplay between Phosphatidylinositol 4,5-bisphosphate (PIP2) and the F-actin network. We show that, during the early stage of cell adhesion, PIP2 forms domains within the filopodia membrane. We studied these domains alongside cell spreading and observed that these very closely follow the actin tread-milling. We show that this mechanism is associated with an active transport of PIP2 rich organelles from the cell perinuclear area to the edge, along actin fibers. Finally, mapping other phospholipids and membrane components we observed that the PIP2 domains formation is correlated with sphingosine and cholesterol rafts.
authors.affiliation: 1] Department of Chemistry, University College London, London, United Kingdom [2] The MRC/UCL Centre for Molecular and Medical Virology, University College London, London, United Kingdom.
authors.forename: Luca
authors.initials: L
authors.lastname: Chierico
authors.affiliation: 1] Department of Chemistry, University College London, London, United Kingdom [2] The MRC/UCL Centre for Molecular and Medical Virology, University College London, London, United Kingdom.
authors.forename: Adrian S
authors.initials: AS
authors.lastname: Joseph
authors.affiliation: Biocompatibles UK Ltd., Farnham Business Park, Weydon Lane, Farnham, United Kingdom.
authors.forename: Andrew L
authors.initials: AL
authors.lastname: Lewis
authors.affiliation: 1] Department of Chemistry, University College London, London, United Kingdom [2] The MRC/UCL Centre for Molecular and Medical Virology, University College London, London, United Kingdom.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Battaglia
mesh_terms: D016475:3T3 Cells; D008841:Actin Cytoskeleton / Q000378:metabolism*; D051376:Actin-Related Protein 2-3 Complex / Q000037:antagonists & inhibitors; D000199:Actins / Q000378:metabolism*; D000818:Animals; D001693:Biological Transport, Active; D002448:Cell Adhesion / Q000502:physiology*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism* / Q000502:physiology; D002465:Cell Movement / Q000502:physiology; D002784:Cholesterol / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D013110:Sphingosine / Q000378:metabolism; D013499:Surface Properties
chemical_list: D051376:Actin-Related Protein 2-3 Complex; D000199:Actins; D019269:Phosphatidylinositol 4,5-Diphosphate; D002784:Cholesterol; D013110:Sphingosine