Physical techniques for the study of exocytosis in isolated cells

• Published in: Biochimie Vol. 80; no. 5; pp. 371 - 377
• Main Authors: Henry, J.P., Darchen, F., Cribier, S.
• Format: Journal Article
• Language: English
• Published: France Elsevier Masson SAS 01-05-1998
• Subjects: amperometry; Animals; Biophysics - methods; Cell Fusion; Cell Membrane - physiology; Electrophysiology - methods; evanescent wave microscopy; exocytosis; Exocytosis - physiology; Fluorescence; membrane fusion; Microscopy, Fluorescence - methods; Optics and Photonics; patch-clamp; Patch-Clamp Techniques; amperometry; GFP, green fluorescent protein; AFM, atomic force microscope; evanescent wave microscopy; CALL, chromophore-assisted laser inactivation; patch-clamp; 5-HT, serotonin; exocytosis; membrane fusion; HA protein, influenza hemagglutinin
• ISSN: 0300-9084; 1638-6183
• DOI: 10.1016/S0300-9084(00)80005-X

Membrane traffic is an important aspect of cell biology which implies shuttle vesicles and multiple binding/fusion events. In spite of rapid progress at the biochemical level, the mechanism of fusion is still not understood. A detailed physical description of the phenomenon is possible at the level of the plasma membrane where secretory vesicles fuse with the cell membrane, a process known as exocytosis. This process is specially active in neurons (release of neurotransmitter) and in endocrine cells (release of hormones), where exocytosis is tightly regulated. Among the biophysical techniques developed, cell membrane capacitance measurements by the technique of patch-clamp and amperometry of the oxidizable secretory products have resulted in interesting information. These techniques have described the initial fusion pore, its fluctuations, the efflux of material through the pore and its irreversible expansion. Optical techniques, using bioluminescent and fluorescent probes are also in progress. For instance, the dye FM 1–43 binds to but is not translocated through biological membranes and it has been used to measure membrane surface, as done by capacitance measurement. Evanescent wave fluorescence microscopy has been recently introduced to analyse the behaviour of secretory granules in the vicinity of the plasma membrane.