Analysis of primary cilia in directional cell migration in fibroblasts

Søren Tvorup Christensen; Iben Veland; Albrecht Schwab; Michael Cammer; Peter Satir

doi:10.1016/B978-0-12-397944-5.00003-1

Analysis of primary cilia in directional cell migration in fibroblasts

Søren Tvorup Christensen, Iben Veland, Albrecht Schwab, Michael Cammer, Peter Satir

Cell Biology and Physiology

14 Citations (Scopus)

Abstract

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.

Original language	English
Book series	Methods in Enzymology
Volume	525
Pages (from-to)	45-58
Number of pages	14
ISSN	0076-6879
DOIs	https://doi.org/10.1016/B978-0-12-397944-5.00003-1
Publication status	Published - 2013

Access to Document

10.1016/B978-0-12-397944-5.00003-1

Analysis of Primary Cilia in Directional Cell Migration in FibroblastsFinal published version, 1.18 MB

Cite this

@article{0a388bc6208a47f18d915f57aad196f5,

title = "Analysis of primary cilia in directional cell migration in fibroblasts",

abstract = "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.",

author = "Christensen, {S{\o}ren Tvorup} and Iben Veland and Albrecht Schwab and Michael Cammer and Peter Satir",

note = "Chapter Three in Cilia, Part B",

year = "2013",

doi = "10.1016/B978-0-12-397944-5.00003-1",

language = "English",

volume = "525",

pages = "45--58",

journal = "Methods in Enzymology",

issn = "0076-6879",

publisher = "Academic Press",

}

TY - JOUR

T1 - Analysis of primary cilia in directional cell migration in fibroblasts

AU - Christensen, Søren Tvorup

AU - Veland, Iben

AU - Schwab, Albrecht

AU - Cammer, Michael

AU - Satir, Peter

N1 - Chapter Three in Cilia, Part B

PY - 2013

Y1 - 2013

N2 - 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.

AB - 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.

U2 - 10.1016/B978-0-12-397944-5.00003-1

DO - 10.1016/B978-0-12-397944-5.00003-1

M3 - Journal article

C2 - 23522464

SN - 0076-6879

VL - 525

SP - 45

EP - 58

JO - Methods in Enzymology

JF - Methods in Enzymology

ER -