The Journal of Cell Biology, Vol 107, 2117-2123, Copyright © 1988 by The Rockefeller University Press

ARTICLES

The dynamics of exocytosis in human neutrophils

O Nusse and M Lindau
Department of Physics, Freie Universitat Berlin, Germany.

We have investigated the dynamics of exocytosis in single human neutrophils. The increase of membrane area associated with granule fusion was followed by time-resolved patch-clamp capacitance measurements. Intracellular application of 20 microM guanosine-5'-O(3- thiotriphosphate) (GTP gamma S) in the presence of 2.5 mM ATP stimulated exocytosis and led to an increase of membrane capacitance from 3.0 to integral of 8.4 pF corresponding to a 540 micron 2 increase of membrane area. This capacitance change is very close to the value expected from morphological data if all primary and secondary granules fuse with the plasma membrane. High resolution measurements revealed stepwise capacitance changes corresponding to the fusion of individual granules. GTP gamma S-stimulated exocytosis did not require pretreatment with cytochalasin B and the amplitude was independent of the intracellular-free calcium concentration between 10 nM and integral of 2.5 microM. In the absence of GTP gamma S elevation of intracellular- free calcium concentration to the micromolar range led to the fusion of only a limited number of granules. Degranulation stimulated with GTP gamma S started after a lag phase of 2-7 min and was usually complete within 5-20 min. The time course was affected by the intracellular ATP and calcium concentration. Exocytosis was markedly accelerated by pretreatment with cytochalasin B. Our results demonstrate that the final steps leading to primary and secondary granule fusion are controlled by a guanine nucleotide-binding protein and do not require an elevation of intracellular calcium. Calcium and other factors are, however, involved in the regulation having pronounced effects on the dynamics of exocytosis.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

• 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]  
• Dewitt, S., Hallett, M. (2007). Leukocyte membrane "expansion": a central mechanism for leukocyte extravasation. J. Leukoc. Biol. 81: 1160-1164 [Abstract] [Full Text]  
• Liu, D., Xu, L., Yang, F., Li, D., Gong, F., Xu, T. (2005). Rapid biogenesis and sensitization of secretory lysosomes in NK cells mediated by target-cell recognition. Proc. Natl. Acad. Sci. USA 102: 123-127 [Abstract] [Full Text]  
• Abdel-Latif, D., Steward, M., Macdonald, D. L., Francis, G. A., Dinauer, M. C., Lacy, P. (2004). Rac2 is critical for neutrophil primary granule exocytosis. Blood 104: 832-839 [Abstract] [Full Text]  
• Hartmann, J., Scepek, S., Hafez, I., Lindau, M. (2003). Differential Regulation of Exocytotic Fusion and Granule-Granule Fusion in Eosinophils by Ca2+ and GTP Analogs. J. Biol. Chem. 278: 44929-44934 [Abstract] [Full Text]  
• Decoursey, T. E. (2003). Voltage-Gated Proton Channels and Other Proton Transfer Pathways. Physiol. Rev. 83: 475-579 [Abstract] [Full Text]  
• Burgoyne, R. D., Morgan, A. (2003). Secretory Granule Exocytosis. Physiol. Rev. 83: 581-632 [Abstract] [Full Text]  
• Theander, S., Lew, D. P., Nusse, O. (2002). Granule-specific ATP requirements for Ca2+-induced exocytosis in human neutrophils. Evidence for substantial ATP-independent release. J. Cell Sci. 115: 2975-2983 [Abstract] [Full Text]  
• Kilic, G., Angleson, J. K, Cochilla, A. J, Nussinovitch, I., Betz, W. J (2001). Sustained stimulation of exocytosis triggers continuous membrane retrieval in rat pituitary somatotrophs. J. Physiol. 532: 771-783 [Abstract] [Full Text]  
• Serrander, L., Skarman, P., Rasmussen, B., Witke, W., Lew, D. P., Krause, K.-H., Stendahl, O., Nu{beta}e, O. (2000). Selective Inhibition of IgG-Mediated Phagocytosis in Gelsolin-Deficient Murine Neutrophils. J. Immunol. 165: 2451-2457 [Abstract] [Full Text]  
• Schmoranzer, J., Goulian, M., Axelrod, D., Simon, S. M. (2000). Imaging Constitutive Exocytosis with Total Internal Reflection Fluorescence Microscopy. JCB 149: 23-32 [Abstract] [Full Text]  
• Becker, K., Hart, N. (1999). Reorganization of filamentous actin and myosin-II in zebrafish eggs correlates temporally and spatially with cortical granule exocytosis. J. Cell Sci. 112: 97-110 [Abstract]  
• Nusse, O., Serrander, L., Foyouzi-Youssefi, R., Monod, A., Lew, D. P., Krause, K.-H. (1997). Store-operated Ca2+ Influx and Stimulation of Exocytosis in HL-60 Granulocytes. J. Biol. Chem. 272: 28360-28367 [Abstract] [Full Text]  
• Borregaard, N., Cowland, J. B. (1997). Granules of the Human Neutrophilic Polymorphonuclear Leukocyte. Blood 89: 3503-3521 [Full Text]  
• Borovikov, Y., Norman, J., Price, L., Weeds, A, Koffer, A (1995). Secretion from permeabilised mast cells is enhanced by addition of gelsolin: contrasting effects of endogenous gelsolin. J. Cell Sci. 108: 657-666 [Abstract]  
• Lindau, M, Nusse, O, Bennett, J, Cromwell, O (1993). The membrane fusion events in degranulating guinea pig eosinophils. J. Cell Sci. 104: 203-210 [Abstract]  
• Di, A., Krupa, B., Nelson, D. J. (2001). Calcium-G Protein Interactions in the Regulation of Macrophage Secretion. J. Biol. Chem. 276: 37124-37132 [Abstract] [Full Text]  

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