1 Cell Biology and Physiology, Department of Biology, Faculty of Science, Københavns Universitet2 Kveiborg Group, BRIC, Faculty of Health and Medical Sciences, Københavns Universitet3 Münster University4 New York University5 Albert Einstein College of Medicine6 Cell Biology and Physiology, Department of Biology, Faculty of Science, Københavns Universitet7 Kveiborg Group, BRIC, Faculty of Health and Medical Sciences, Københavns Universitet
Early studies of migrating fibroblasts showed that primary cilia orient in front of the nucleus and point toward the leading edge. Recent work has shown that primary cilia coordinate a series of signaling pathways critical to fibroblast cell migration during development and in wound healing. In particular, platelet-derived growth factor receptor alpha (PDGFRα) is compartmentalized to the primary cilium to activate signaling pathways that regulate reorganization of the cytoskeleton required for lamellipodium formation and directional migration in the presence of a specific ligand gradient. We summarize selected methods in analyzing ciliary function in directional cell migration, including immunofluorescence microscopy, scratch assay, and chemotaxis assay by micropipette addition of PDGFRα ligands to cultures of fibroblasts. These methods should be useful not only in studying cell migration but also more generally in delineating response pathways in cells with primary cilia.