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© The Rockefeller University Press, 0021-9525/2000/5/697/ $5.00
The Journal of Cell Biology, Volume 149, Number 3, May 1, 2000 697-706

Original Article

Focal Exocytosis of VAMP3-containing Vesicles at Sites of Phagosome Formation

Lydia Bajnoa, Xiao-Rong Penga, Alan D. Schreiberb, Hsiao-Ping Moorec, William S. Trimblea, and Sergio Grinsteina
a Cell Biology Programme, Research Institute, The Hospital for Sick Children and Department of Biochemistry, University of Toronto, Toronto, M5G 1X8 Ontario, Canada
b Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104
c Department of Cell Biology, University of California, Berkeley, California 94720

Correspondence to: Sergio Grinstein, Cell Biology Program, Hospital for Sick Children, 555 University Avenue, Toronto, M5G 1X8 Canada. Tel:(416) 813-5727 Fax:(416) 813-5028 E-mail:sga{at}sickkids.on.ca.

Phagocytosis involves the receptor-mediated extension of plasmalemmal protrusions, called pseudopods, which fuse at their tip to engulf a particle. Actin polymerizes under the nascent phagosome and may propel the protrusion of pseudopods. Alternatively, membrane extension could result from the localized insertion of intracellular membranes into the plasmalemma next to the particle. Here we show focal accumulation of VAMP3-containing vesicles, likely derived from recycling endosomes, in the vicinity of the nascent phagosome. Using green fluorescent protein (GFP) as both a fluorescent indicator and an exofacial epitope tag, we show that polarized fusion of VAMP3 vesicles precedes phagosome sealing. It is therefore likely that targeted delivery of endomembranes contributes to the elongation of pseudopods. In addition to mediating pseudopod formation, receptor-triggered focal secretion of endosomes may contribute to polarized membrane extension in processes such as lamellipodial elongation or chemotaxis.

Key Words: cellubrevin, phagocytosis, macrophage, recycling endosomes, GFP


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