Neutrophil microtubules suppress polarity and enhance directional migration.
How do microtubules, which maintain and direct polarity of many eukaryotic cells, regulate polarity of blood neutrophils? In sharp contrast to most cells, disrupting a neutrophil's microtubule network with nocodazole causes it to polarize and migrate [Niggli, V. (2003) J. Cell Sci. 116, 813-822]. Nocodazole induces the same responses in differentiated HL-60 cells, a model neutrophil cell line, and reduces their chemotactic prowess by causing them to pursue abnormally circuitous paths in migrating toward a stationary point source of an attractant, f-Met-Leu-Phe (fMLP). The chemotactic defect stems from dramatic nocodazole-induced imbalance between the divergent, opposed fMLP-induced "backness" and "frontness" signals responsible for neutrophil polarity. Nocodazole (i) stimulates backness by increasing Rho- and actomyosin-dependent contractility, as reported by Niggli, and also (ii) impairs fMLP-dependent frontness: pseudopods are flatter, contain less F-actin, and show decreased membrane translocation of PH-Akt-GFP, a fluorescent marker for 3'-phosphoinositide lipids. Inhibiting backness with a pharmacologic inhibitor of a Rho-dependent kinase substantially reverses nocodazole's effects on chemotaxis, straightness of migration paths, morphology, and PH-Akt-GFP translocation. Thus, microtubules normally balance backness vs. frontness signals, preventing backness from reducing the strength of pseudopods and from impairing directional migration.
Xu
Jingsong
J
Department of Cellular and Molecular Pharmacology and Cardiovascular Research Institute, University of California, San Francisco, CA 94143-0450, USA.
Wang
Fei
F
Van Keymeulen
Alexandra
A
Rentel
Maike
M
Bourne
Henry R
HR
D000970:Antineoplastic Agents / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement; D002633:Chemotaxis; D018076:DNA, Complementary / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018922:HL-60 Cells; D006801:Humans; D008870:Microtubules / Q000378:metabolism*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000378:metabolism*; D015739:Nocodazole / Q000494:pharmacology; D021381:Protein Transport; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection
D000970:Antineoplastic Agents; D018076:DNA, Complementary; D049452:Green Fluorescent Proteins; D009240:N-Formylmethionine Leucyl-Phenylalanine; D015739:Nocodazole