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Fig. 5. Secretagogues induced fast reorganisations in F-actin cytoskeleton in chromaffin cells. Cells were stimulated by addition of acetylcholine (10 µM) to the basal bathing medium and the changes in the F-actin cytoskeleton studied by acquiring transmitted light images of the entire cell at 1 Hz. (A) Frames separated by 10-second intervals after secretagogue addition. The formation of clear discontinuities in the subcortical structures is visible. (B) Measurements of optical density in a ROI located over one of the detected disruptions for 2-second intervals, showed that the formation of these subcortical patches within a few seconds was fast enough to encompass secretion measured by the increase in the cell perimeter (D). (C) In contrast with these rapid changes occurring in the cell periphery, reorganisation of intracellular structures forming polygons increased the open space in their interior. This process takes tens of seconds to fully develop as shown in the images taken at 10-second intervals. Bar, 10 µm (A); 1 µm (B,C).
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