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¹ Laboratory of Cellular Biophysics, Rockefeller University, New York, NY 10021
² Section of Microbial Pathogenesis and Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520

Address correspondence to Sanford M. Simon, Laboratory of Cellular Biophysics, Rockefeller University, Box 304, 1230 York Ave., New York, NY 10021. Tel.: (212) 327-8130. Fax: (212) 327-7543. E-mail: simon{at}rockefeller.edu

Similar to its role in secretory cells, calcium triggers exocytosis in nonsecretory cells. This calcium-dependent exocytosis is essential for repair of membrane ruptures. Using total internal reflection fluorescence microscopy, we observed that many organelles implicated in this process, including ER, post-Golgi vesicles, late endosomes, early endosomes, and lysosomes, were within 100 nm of the plasma membrane (in the evanescent field). However, an increase in cytosolic calcium led to exocytosis of only the lysosomes. The lysosomes that fused were predominantly predocked at the plasma membrane, indicating that calcium is primarily responsible for fusion and not recruitment of lysosomes to the cell surface.

Key Words: TIR-FM; CD63; dextran; VAMP7; Synaptotagmin VII

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