Summary
This cell-free assay for exocytosis is particularly useful when spatial information about exocytotic sites and biochemical access to the plasma membrane within less than a minute is required. It is based on the study of plasma membrane lawns from secretory cells exhibiting secretory granules filled with neuropeptide Y-green fluorescent protein (NPY-GFP). The sample is prepared by subjecting NPY-GFP-expressing cells to a brief ultrasound pulse, leaving behind a basal, flat plasma membrane with fluorescent attached secretory organelles. These sheets can then be incubated in defined solutions with the benefit that complete solution changes can be achieved in less than 1 min. Individual secretory granules are monitored in the docked state and during exocytosis by video microscopy.
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Acknowledgments
I would like to thank Dr. Philip Holroyd for critical comments on the manuscript and providing images of membrane sheets.
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Lang, T. (2008). Imaging Ca2+-Triggered Exocytosis of Single Secretory Granules on Plasma Membrane Lawns from Neuroendocrine Cells. In: Ivanov, A.I. (eds) Exocytosis and Endocytosis. Methods in Molecular Biology, vol 440. Humana Press. https://doi.org/10.1007/978-1-59745-178-9_4
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DOI: https://doi.org/10.1007/978-1-59745-178-9_4
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