The Journal of Cell Biology, Vol 98, 1265-1271, Copyright © 1984 by The Rockefeller University Press

ARTICLES

Chemotactic peptide modulation of actin assembly and locomotion in neutrophils

TH Howard and WH Meyer

To determine the relationship between the state of actin polymerization in neutrophils and the formyl-methionyl-leucyl-phenylalanine (fMLP)- induced changes in the locomotive behavior of neutrophils, the mean rate of locomotion (mROL), the percent G-actin, and the relative F- actin content of neutrophils were determined. The mROL was quantified by analysis of the locomotion of individual cells; the percentage of total actin as G-actin was measured by DNase I inhibition; and the F- actin was determined by fluorescence-activated cell sorter (FACS) analysis of nitrobenzoxadiazol (NBD)-phallacidin-stained neutrophils. Neutrophils stimulated with fMLP exhibit a change in their mROL that is biphasic and dose dependent. The mROL of neutrophils exposed to 10(-8) M fMLP, the KD, is 11.9 +/- 2.0 micron/min (baseline control 6.2 +/- 1.0 micron/min). At 10(-6) M fMLP, the mROL returns to baseline levels. Stimulation of neutrophils with fMLP also induces action polymerization. Evidence for actin polymerization includes a 26.5% reduction in G-actin and a twofold increase in the amount of NBD- phallacidin staining of cells as determined by FACS analysis. The NBD- phallacidin staining is not due to phagocytosis, is inhibited by phalloidin, requires cell permeabilization, and is saturable at NBD- phallacidin concentrations greater than 10(-7)M. The fMLP-induced increase in NBD-phallacidin staining occurs rapidly (less than 2 min), is temperature dependent, and is not due to cell aggregation. Since NBD- phallacidin binds specifically to F-actin, the increase in fluorescent staining of cells likely reflects an increase in the F-actin content of fMLP-stimulated cells. FACS analysis of NBD-phallacidin-stained cells shows that the relative F-actin content of neutrophils stimulated with 10(-11)-10(-8)M fMLP increases twofold and remains increased at concentrations greater than 10(-8)M fMLP. Therefore, the fMLP-induced increase in F-actin content of neutrophils as determined by FACS analysis of NBD-phallacidin-stained cells coincides with a decrease in G-actin and correlates with increased mROL of neutrophils under some (10(-11)-10(-8)M fMLP) but not all (greater than 10(-8)M fMLP) conditions of stimulation. Quantification of the F-actin content of nonmuscle cells by FACS analysis of NBD-phallacidin-stained cells may allow rapid assessment of the state of actin polymerization and correlation of that state with the motile behavior of nonmuscle cells.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

• Basu, S., Broxmeyer, H. E. (2009). CCR5 Ligands Modulate CXCL12-Induced Chemotaxis, Adhesion, and Akt Phosphorylation of Human Cord Blood CD34+ Cells. J. Immunol. 183: 7478-7488 [Abstract] [Full Text]  
• Eckels, P. C., Banerjee, A., Moore, E. E., McLaughlin, N. J. D., Gries, L. M., Kelher, M. R., England, K. M., Gamboni-Robertson, F., Khan, S. Y., Silliman, C. C. (2009). Amantadine inhibits platelet-activating factor induced clathrin-mediated endocytosis in human neutrophils. Am. J. Physiol. Cell Physiol. 297: C886-C897 [Abstract] [Full Text]  
• Reddy, R. C., Narala, V. R., Keshamouni, V. G., Milam, J. E., Newstead, M. W., Standiford, T. J. (2008). Sepsis-induced inhibition of neutrophil chemotaxis is mediated by activation of peroxisome proliferator-activated receptor-{gamma}. Blood 112: 4250-4258 [Abstract] [Full Text]  
• Atkin, L. (2008). Rosiglitazone-induced SERCA2b inhibition: implications for monocyte cytoskeletal remodelling and diabetic microangiopathy. Bioscience Horizons 1: 1-8 [Abstract] [Full Text]  
• Draberova, L., Shaik, G. M., Volna, P., Heneberg, P., Tumova, M., Lebduska, P., Korb, J., Draber, P. (2007). Regulation of Ca2+ Signaling in Mast Cells by Tyrosine-Phosphorylated and Unphosphorylated Non-T Cell Activation Linker. J. Immunol. 179: 5169-5180 [Abstract] [Full Text]  
• Bourguignon, L. Y. W., Peyrollier, K., Gilad, E., Brightman, A. (2007). Hyaluronan-CD44 Interaction with Neural Wiskott-Aldrich Syndrome Protein (N-WASP) Promotes Actin Polymerization and ErbB2 Activation Leading to beta-Catenin Nuclear Translocation, Transcriptional Up-regulation, and Cell Migration in Ovarian Tumor Cells. J. Biol. Chem. 282: 1265-1280 [Abstract] [Full Text]  
• Kobayashi, Y., Ueki, S., Mahemuti, G., Chiba, T., Oyamada, H., Saito, N., Kanda, A., Kayaba, H., Chihara, J. (2005). Physiological Levels of 15-Deoxy-{Delta}12,14-Prostaglandin J2 Prime Eotaxin-Induced Chemotaxis on Human Eosinophils through Peroxisome Proliferator-Activated Receptor-{gamma} Ligation. J. Immunol. 175: 5744-5750 [Abstract] [Full Text]  
• Basu, S., Broxmeyer, H. E. (2005). Transforming growth factor-{beta}1 modulates responses of CD34+ cord blood cells to stromal cell-derived factor-1/CXCL12. Blood 106: 485-493 [Abstract] [Full Text]  
• Gutzmer, R., Diestel, C., Mommert, S., Kother, B., Stark, H., Wittmann, M., Werfel, T. (2005). Histamine H4 Receptor Stimulation Suppresses IL-12p70 Production and Mediates Chemotaxis in Human Monocyte-Derived Dendritic Cells. J. Immunol. 174: 5224-5232 [Abstract] [Full Text]  
• Dhawan, J., Helfman, D. M. (2004). Modulation of acto-myosin contractility in skeletal muscle myoblasts uncouples growth arrest from differentiation. J. Cell Sci. 117: 3735-3748 [Abstract] [Full Text]  
• Kimura, T., Boehmler, A. M., Seitz, G., Kuci, S., Wiesner, T., Brinkmann, V., Kanz, L., Mohle, R. (2004). The sphingosine 1-phosphate receptor agonist FTY720 supports CXCR4-dependent migration and bone marrow homing of human CD34+ progenitor cells. Blood 103: 4478-4486 [Abstract] [Full Text]  
• Kutsuna, H., Suzuki, K., Kamata, N., Kato, T., Hato, F., Mizuno, K., Kobayashi, H., Ishii, M., Kitagawa, S. (2004). Actin reorganization and morphological changes in human neutrophils stimulated by TNF, GM-CSF, and G-CSF: the role of MAP kinases. Am. J. Physiol. Cell Physiol. 286: C55-C64 [Abstract] [Full Text]  
• Gutzmer, R., Langer, K., Mommert, S., Wittmann, M., Kapp, A., Werfel, T. (2003). Human Dendritic Cells Express the IL-18R and Are Chemoattracted to IL-18. J. Immunol. 171: 6363-6371 [Abstract] [Full Text]  
• Draberova, L., Dudkova, L., Boubelik, M., Tolarova, H., Smid, F., Draber, P. (2003). Exogenous Administration of Gangliosides Inhibits Fc{epsilon}RI-Mediated Mast Cell Degranulation by Decreasing the Activity of Phospholipase C{gamma}. J. Immunol. 171: 3585-3593 [Abstract] [Full Text]  
• Miller, Y. I., Worrall, D. S., Funk, C. D., Feramisco, J. R., Witztum, J. L. (2003). Actin Polymerization in Macrophages in Response to Oxidized LDL and Apoptotic Cells: Role of 12/15-Lipoxygenase and Phosphoinositide 3-Kinase. Mol. Biol. Cell 14: 4196-4206 [Abstract] [Full Text]  
• Rabin, R. L., Alston, M. A., Sircus, J. C., Knollmann-Ritschel, B., Moratz, C., Ngo, D., Farber, J. M. (2003). CXCR3 Is Induced Early on the Pathway of CD4+ T Cell Differentiation and Bridges Central and Peripheral Functions. J. Immunol. 171: 2812-2824 [Abstract] [Full Text]  
• Schmitt, N., Chene, L., Boutolleau, D., Nugeyre, M.-T., Guillemard, E., Versmisse, P., Jacquemot, C., Barre-Sinoussi, F., Israel, N. (2003). Positive Regulation of CXCR4 Expression and Signaling by Interleukin-7 in CD4+ Mature Thymocytes Correlates with Their Capacity To Favor Human Immunodeficiency X4 Virus Replication. J. Virol. 77: 5784-5793 [Abstract] [Full Text]  
• Fuhler, G. M., Drayer, A. L., Vellenga, E. (2003). Decreased phosphorylation of protein kinase B and extracellular signal-regulated kinase in neutrophils from patients with myelodysplasia. Blood 101: 1172-1180 [Abstract] [Full Text]  
• Copeland, J. W., Treisman, R. (2002). The Diaphanous-related Formin mDia1 Controls Serum Response Factor Activity through its Effects on Actin Polymerization. Mol. Biol. Cell 13: 4088-4099 [Abstract] [Full Text]  
• Rescher, U., Danielczyk, A., Markoff, A., Gerke, V. (2002). Functional Activation of the Formyl Peptide Receptor by a New Endogenous Ligand in Human Lung A549 Cells. J. Immunol. 169: 1500-1504 [Abstract] [Full Text]  
• Geneste, O., Copeland, J. W., Treisman, R. (2002). LIM kinase and Diaphanous cooperate to regulate serum response factor and actin dynamics. JCB 157: 831-838 [Abstract] [Full Text]  
• Hua, J., Sakamoto, K., Nagaoka, I. (2002). Inhibitory actions of glucosamine, a therapeutic agent for osteoarthritis, on the functions of neutrophils. J. Leukoc. Biol. 71: 632-640 [Abstract] [Full Text]  
• Ginzberg, H. H., Cherapanov, V., Dong, Q., Cantin, A., McCulloch, C. A. G., Shannon, P. T., Downey, G. P. (2001). Neutrophil-mediated epithelial injury during transmigration: role of elastase. Am. J. Physiol. Gastrointest. Liver Physiol. 281: G705-G717 [Abstract] [Full Text]  
• Bautz, F., Denzlinger, C., Kanz, L., Mohle, R. (2001). Chemotaxis and transendothelial migration of CD34+ hematopoietic progenitor cells induced by the inflammatory mediator leukotriene D4 are mediated by the 7-transmembrane receptor CysLT1. Blood 97: 3433-3440 [Abstract] [Full Text]  
• Li, Y., Zhang, Q., Aaron, R., Hilliard, L., Howard, T. H. (2000). LSP1 modulates the locomotion of monocyte-differentiated U937 cells. Blood 96: 1100-1105 [Abstract] [Full Text]  
• Tezel, G., Wax, M. B. (2000). The Mechanisms of hsp27 Antibody-Mediated Apoptosis in Retinal Neuronal Cells. J. Neurosci. 20: 3552-3562 [Abstract] [Full Text]  
• Rengan, R., Ochs, H. D., Sweet, L. I., Keil, M. L., Gunning, W. T., Lachant, N. A., Boxer, L. A., Omann, G. M. (2000). Actin cytoskeletal function is spared, but apoptosis is increased, in WAS patient hematopoietic cells. Blood 95: 1283-1292 [Abstract] [Full Text]  
• Anderson, S., Hotchin, N., Nash, G. (2000). Role of the cytoskeleton in rapid activation of CD11b/CD18 function and its subsequent downregulation in neutrophils. J. Cell Sci. 113: 2737-2745 [Abstract]  
• Sotsios, Y., Whittaker, G. C., Westwick, J., Ward, S. G. (1999). The CXC Chemokine Stromal Cell-Derived Factor Activates a Gi-Coupled Phosphoinositide 3-Kinase in T Lymphocytes. J. Immunol. 163: 5954-5963 [Abstract] [Full Text]  
• Feltis, B. A., Kim, A. S., Kinneberg, K. M., Lyerly, D. L., Wilkins, T. D., Erlandsen, S. L., Wells, C. L. (1999). Clostridium difficile Toxins May Augment Bacterial Penetration of Intestinal Epithelium. Arch Surg 134: 1235-1242 [Abstract] [Full Text]  
• Kim, C. H., Qu, C.-K., Hangoc, G., Cooper, S., Anzai, N., Feng, G.-S., Broxmeyer, H. E. (1999). Abnormal Chemokine-Induced Responses of Immature and Mature Hematopoietic Cells from Motheaten Mice Implicate the Protein Tyrosine Phosphatase Shp-1 in Chemokine Responses. JEM 190: 681-690 [Abstract] [Full Text]  
• Masuyama, J.-i., Yoshio, T., Suzuki, K., Kitagawa, S., Iwamoto, M., Kamimura, T., Hirata, D., Takeda, A., Kano, S., Minota, S. (1999). Characterization of the 4C8 Antigen Involved in Transendothelial Migration of CD26hi T Cells after Tight Adhesion to Human Umbilical Vein Endothelial Cell Monolayers. JEM 189: 979-990 [Abstract] [Full Text]  
• Wells, C. L., Jechorek, R. P., Kinneberg, K. M., Debol, S. M., Erlandsen, S. L. (1999). The Isoflavone Genistein Inhibits Internalization of Enteric Bacteria by Cultured Caco-2 and HT-29 Enterocytes. J. Nutr. 129: 634-640 [Abstract] [Full Text]  
• Leavey, P. J., Sellins, K. S., Thurman, G., Elzi, D., Hiester, A., Silliman, C. C., Zerbe, G., Cohen, J. J., Ambruso, D. R. (1998). In Vivo Treatment With Granulocyte Colony-Stimulating Factor Results in Divergent Effects on Neutrophil Functions Measured In Vitro. Blood 92: 4366-4374 [Abstract] [Full Text]  
• Lippert, U., Artuc, M., Grutzkau, A., Moller, A., Kenderessy-Szabo, A., Schadendorf, D., Norgauer, J., Hartmann, K., Schweitzer-Stenner, R., Zuberbier, T., Henz, B. M., Kruger-Krasagakes, S. (1998). Expression and Functional Activity of the IL-8 Receptor Type CXCR1 and CXCR2 on Human Mast Cells. J. Immunol. 161: 2600-2608 [Abstract] [Full Text]  
• Wells, C. L., van de Westerlo, E. M. A., Jechorek, R. P., Haines, H. M., Erlandsen, S. L. (1998). Cytochalasin-Induced Actin Disruption of Polarized Enterocytes Can Augment Internalization of Bacteria. Infect. Immun. 66: 2410-2419 [Abstract] [Full Text]  
• Banan, A., McCormack, S. A., Johnson, L. R. (1998). Polyamines are required for microtubule formation during gastric mucosal healing. Am. J. Physiol. Gastrointest. Liver Physiol. 274: G879-G885 [Abstract] [Full Text]  
• Bleul, C. C., Schultze, J. L., Springer, T. A. (1998). B Lymphocyte Chemotaxis Regulated in Association with Microanatomic Localization, Differentiation State, and B Cell Receptor Engagement. JEM 187: 753-762 [Abstract] [Full Text]  
• Petering, H., Hochstetter, R., Kimmig, D., Smolarski, R., Kapp, A., Elsner, J. (1998). Cutting Edge: Detection of MCP-4 in Dermal Fibroblasts and Its Activation of the Respiratory Burst in Human Eosinophils. J. Immunol. 160: 555-558 [Abstract] [Full Text]  
• Arcaro, A. (1998). The Small GTP-binding Protein Rac Promotes the Dissociation of Gelsolin from Actin Filaments in Neutrophils. J. Biol. Chem. 273: 805-813 [Abstract] [Full Text]  
• Downey, G. P., Butler, J. R., Tapper, H., Fialkow, L., Saltiel, A. R., Rubin, B. B., Grinstein, S. (1998). Importance of MEK in Neutrophil Microbicidal Responsiveness. J. Immunol. 160: 434-443 [Abstract] [Full Text]  
• Kim, C. H., Broxmeyer, H. E. (1998). In Vitro Behavior of Hematopoietic Progenitor Cells Under the Influence of Chemoattractants: Stromal Cell-Derived Factor-1, Steel Factor, and the Bone Marrow Environment. Blood 91: 100-110 [Abstract] [Full Text]  
• Clunn, G. F., Refson, J. S., Lymn, J. S., Hughes, A. D. (1997). Platelet-Derived Growth Factor ß-Receptors Can Both Promote and Inhibit Chemotaxis in Human Vascular Smooth Muscle Cells. Arterioscler. Thromb. Vasc. Bio. 17: 2622-2629 [Abstract] [Full Text]  
• Machesky, L. M., Hall, A. (1997). Role of Actin Polymerization and Adhesion to Extracellular Matrix in Rac- and Rho-induced Cytoskeletal Reorganization. JCB 138: 913-926 [Abstract] [Full Text]  
• Syrovets, T., Tippler, B., Rieks, M., Simmet, T. (1997). Plasmin Is a Potent and Specific Chemoattractant for Human Peripheral Monocytes Acting Via a Cyclic Guanosine Monophosphate-Dependent Pathway. Blood 89: 4574-4583 [Abstract] [Full Text]  
• Derman, M. P., Toker, A., Hartwig, J. H., Spokes, K., Falck, J. R., Chen, C.-S., Cantley, L. C., Cantley, L. G. (1997). The Lipid Products of Phosphoinositide 3-Kinase Increase Cell Motility through Protein Kinase C. J. Biol. Chem. 272: 6465-6470 [Abstract] [Full Text]  
• Zhou, D., Luini, W., Bernasconi, S., Diomede, L., Salmona, M., Mantovani, A., Sozzani, S. (1995). Phosphatidic Acid and Lysophosphatidic Acid Induce Haptotactic Migration of Human Monocytes. J. Biol. Chem. 270: 25549-25556 [Abstract] [Full Text]  
• Bershadsky, A., Gluck, U, Denisenko, O., Sklyarova, T., Spector, I, Ben-Ze'ev, A (1995). The state of actin assembly regulates actin and vinculin expression by a feedback loop. J. Cell Sci. 108: 1183-1193 [Abstract]  
• Shariff, A, Luna, E. (1992). Diacylglycerol-stimulated formation of actin nucleation sites at plasma membranes. Science 256: 245-247 [Abstract]  
• Ashkenazi, M., Dennison, D.K. (1989). A New Method for Isolation of Salivary Neutrophils and Determination of Their Functional Activity. JDR 68: 1256-1261 [Abstract]  
• Miller, Y. I., Chang, M.-K., Funk, C. D., Feramisco, J. R., Witztum, J. L. (2001). 12/15-Lipoxygenase Translocation Enhances Site-specific Actin Polymerization in Macrophages Phagocytosing Apoptotic Cells. J. Biol. Chem. 276: 19431-19439 [Abstract] [Full Text]  
• Shive, M. S., Salloum, M. L., Anderson, J. M. (2000). Shear stress-induced apoptosis of adherent neutrophils: A mechanism for persistence of cardiovascular device infections. Proc. Natl. Acad. Sci. USA 97: 6710-6715 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents