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    Measurement of the Dynamics of Exocytosis and Vesicle Retrieval at Cell Populations Using a Quartz Crystal Microbalance

    Year: 2001

    Journal: Analytical Chemistry 2001, 73, 5805-5811, 20100827

    Authors: Cans A-S, Höök F., Shupliakov O., Ewing A.G., Eriksson P.S., Brodin L., Orwar O.

    Last authors: Owe Orwar

    Organizations: Department of Chemistry, Göteborg University, S-41296 Göteborg, Sweden Department of Applied Physics, Chalmers University of Technology and Göteborg University, S-41296 Göteborg, Sweden Department of Chemistry, Penn State University, University Park, Pennsylvania, USA Institute of Clinical Neuroscience, Sahlgrenska University Hospital, Göteborg University, S-41345 Göteborg, Sweden Department of Neuroscience, The Nobel Institute for Neurophysiology, Karolinska Institute, Stockholm, Sweden Department of Physical Chemistry and Microtechnology Center, Chalmers University of Technology, S-41296 Göteborg, Sweden

    Country: Sweden

    The quartz crystal microbalance-dissipation technique (QCM-D) is used in two different measurement strategies to monitor the mass change and rigidity of populations of excitable cells during exocytosis and subsequent retrieval of dense-core vesicles. Two cell lines, NG 108-15 and PC 12, were grown to confluence on piezoelectric quartz crystals and were examined separately to demonstrate differences in release and retrieval with cells of different morphology, size, and number of dense-core vesicles. Stimulating the cells to exocytosis with media containing an elevated potassium concentration resulted in an in-crease in the frequency response corresponding to loss of mass from the cells owing to release of vesicles. In Ca2+ free media, the response was completely abolished. The amplitude and peak area in the frequency response corresponding to mass change with stimulated release was larger for PC 12 cells than for NG 108-15 cells, whereas the initial rate constants for the frequency responses were similar. The data suggest (1) that a greater number and larger size of vesicles in PC 12 cells results in a greater amount of release from these cells vs NG 108-15 cells, (2) the recycling of vesicles utilizes similar fusion/retrieval mechanisms in both cell types, (3) that the control of excess retrieval might be related to the number and size of released vesicles, and (4) that measured retrieval has a rapid onset, masking exocytosis and implying a rapid retrieval mechanism in the early stages of release. These results demonstrate that measurements of complex dynamic processes relating to dense-core vesicle release and retrieval can be simultaneously accomplished using the QCM-D technique.