Changes in actin network during calcium-induced exocytosis in permeabilized GH3 cells: Calcium directly regulates F-actin disassembly

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Abstract

Using digitonin-permeabilized GH3 cells, we investigated both the release of prolactin (PRL) and changes in the cytoskeleton. We determined that permeabilized GH3 cells released PRL in a dose-dependent manner upon addition of micromolar Ca2+. Phalloidin, a filamentous actin (F-actin) stabilizing agent, inhibited both Ca2+-dependent and -independent PRL release, whereas cytochalasin B, a destabilizing agent, had almost no effect on the release. Observation with a confocal laser scanning microscope revealed that F-actin existed mainly in the cortical region in the quiescent state. Increased cytosolic Ca2+ induced a change in F-actin distribution: F-actin in the cortical region decreased, whereas F-actin inside the cells increased. This change in F-actin distribution was not observed when phalloidin was added. Addition of cytochalasin B induced patchy F-actin spots, but the pattern of the changes of F-actin distribution did not change. The time course of change in F-actin distribution showed that the F-actin network in the cortical region was reduced within 1 min, and Ca2+-dependent release of PRL continued for up to 20 min. These results suggest that the F-actin network near the membrane acts as a barrier to exocytosis and that Ca2+ directly controls the cytoskeletal changes.

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Yoneda, M., Nishizaki, T., Tasaka, K., Kurachi, H., Miyake, A., & Murata, Y. (2000). Changes in actin network during calcium-induced exocytosis in permeabilized GH3 cells: Calcium directly regulates F-actin disassembly. Journal of Endocrinology, 166(3), 677–687. https://doi.org/10.1677/joe.0.1660677

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