The control of cytoskeletal actin and exocytosis in intact and permeabilized adrenal chromaffin cells: role of calcium and protein kinase C

Burgoyne, Robert D.; Morgan, Alan; O’Sullivan, Antony J.

doi:10.1016/0898-6568(89)90051-x

Cited by 101 publications

(62 citation statements)

References 44 publications

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“…Many agents, such as protein kinase C (Burgoyne et al 1989, Nishizaki et al 1992, calmodulin (Burgoyne 1991, Cheek 1991, fodrin (Burgoyne & Cheek 1987, Perrin et al 1987, gelsolin (Bader et al 1986, Trifaró & Vitale 1993, scinderin (Rodríguez et al 1990, Vitale et al 1995, calpactin , and 14-3-3 proteins (Roth & Burgoyne 1995) and GTP-binding proteins (Norman et al 1994, Price et al 1995, have been studied as Ca 2+ target proteins in the regulation of exocytosis, though the exact mechanisms by which Ca 2+ regulates exocytosis remain unclear. Immunocytochemical studies showed that the cortical actin network was disassembled upon nicotine stimulation of bovine adrenal chromaffin cells (Cheek & Burgoyne 1986, Trifaró & Vitale 1993.…”

Section: Discussionmentioning

confidence: 99%

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

Yoneda¹,

Nishizaki²,

Tasaka³

et al. 2000

Journal of Endocrinology

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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 Ca 2+ . Phalloidin, a filamentous actin (F-actin) stabilizing agent, inhibited both Ca 2+ -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 Ca 2+ 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 Ca 2+ -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 Ca 2+ directly controls the cytoskeletal changes.

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Section: Discussionmentioning

confidence: 99%

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

Yoneda¹,

Nishizaki²,

Tasaka³

et al. 2000

Journal of Endocrinology

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“…Once the application of nicotine had ceased, there was no perfusion of medium over the cells. Subsequently, the NIH-3T3 t cells (cells [2][3][4][5][6][7][8][9][10] responded with a rise in [Ca2+]i with a delay of onset that was related to the distance of the NIH-3T3 t cell from the chromaftin cell, such that the delay was greatest in those NIH-3T3' cells that were furthest from the chromattin cell (Table I). This is consistent with ATP being released from the chromaflin cell and then diffusing to the surrounding NIH-3T3' cells.…”

Section: Immunofluorescence Staining With Anti-dbhmentioning

confidence: 99%

Simultaneous measurements of cytosolic calcium and secretion in single bovine adrenal chromaffin cells by fluorescent imaging of fura-2 in cocultured cells.

Cheek

Jackson

O’Sullivan

et al. 1989

The Journal of Cell Biology

122

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Abstract. The cytosolic free calcium concentration ([Ca2+]i) and exocytosis of chromaffin granules were measured simultaneously from single, intact bovine adrenal chromaffin cells using a novel technique involving fluorescent imaging of cocultured cells. Chromaflin cell [Ca2+]i was monitored with fura-2. To simultaneously follow catecholamine secretion, the cells were cocultured with fura-2-1oaded NIH-3T3' cells, a cell line chosen because of their irresponsiveness to chromaffin cell secretagogues but their large Ca 2+ response to ATP, which is coreleased with catecholamine from the chromaffin cells.In response to the depolarizing stimulus nicotine (a potent secretagogue), chromaffin cell [Ca2+]i increased rapidly. At the peak of the response, [Ca2+]i was evenly distributed throughout the cell. This elevation in [Ca2÷]i was followed by a secretory response which originated from the entire surface of the cell.In response to the inositol 1,4,5-trisphosphate (InsP3)-mobilizing agonist angiotensin II (a weak secretagogue), three different responses were observed. Approximately 30% of chromaftin cells showed no rise in [Ca2+]i and did not secrete. About 45% of the cells responded with a large (>200 nM), transient elevation in [Ca2÷]i and no detectable secretory response. The rise in [Ca2÷]i was nonuniform, such that peak [Ca2+]i was often recorded only in one pole of the cell. And finally, ~25 % of cells responded with a similar Ca2+-transient to that described above, but also gave a secretory response. In these cases secretion was polarized, being confined to the pole of the cell in which the rise in [Ca2+]i was greatest. Exocytosis in response to nicotine occurred over the entire surface of the cell, whereas exocytosis due to angiotensin II was polarized, as was confirmed by immunofluorescent localization of dopamine-B-hydroxylase, a chromaflin granule protein that becomes incorporated into the plasma membrane during fusion.These results directly demonstrate, for the first time, that intact chromaffin cells can undergo a large, agonist-induced transient rise in [Ca2+]i without this stimulating secretion and, furthermore, show that the location of exocytosis around the cell can vary depending on the nature of the stimulus. XOCYTOSlS, the process by which intracellular vesicles fuse with the inner surface of the plasma membrane and release their contents into the surrounding medium, is the mechanism underlying the secretion of many physiologically important mediators such as hormones, enzymes, and neurotransmitters. The process is often regulated by an external signal which stimulates release by altering the level of an intracellular second messenger.The pivotal role that Ca 2÷ plays in triggering exocytosis was first noted nearly 30 yr ago with the demonstration that depolarized chromaffin cells would not secrete catecholamines in the absence of extraceUular Ca 2÷ (13). The involvement of Ca 2÷ in exocytosis was advanced by subsequent information obtained from studies with 45Ca 2÷ (12) All reprint requests should be...

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“…Time-lapse-total internal reflection fluorescence (TIRF) microscopy was used to image NPY-mCherry-positive SVs and lifeact-GFP-positive actin in the transfected cells. Stimulation using either Ba 2 þ (2 mM) 21,22 or nicotine (100 mM) led to a coincident albeit transient increase in fluorescence intensity for both lifeact-GFP and NPY-mCherry (Fig. 1a,b, Supplementary Fig.…”

Section: Svs and Cortical Actin Approach The Plasmalemma On Stimulationmentioning

confidence: 99%

Activity-driven relaxation of the cortical actomyosin II network synchronizes Munc18-1-dependent neurosecretory vesicle docking

et al. 2015

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In neurosecretory cells, secretory vesicles (SVs) undergo Ca 2 þ -dependent fusion with the plasma membrane to release neurotransmitters. How SVs cross the dense mesh of the cortical actin network to reach the plasma membrane remains unclear. Here we reveal that, in bovine chromaffin cells, SVs embedded in the cortical actin network undergo a highly synchronized transition towards the plasma membrane and Munc18-1-dependent docking in response to secretagogues. This movement coincides with a translocation of the cortical actin network in the same direction. Both effects are abolished by the knockdown or the pharmacological inhibition of myosin II, suggesting changes in actomyosin-generated forces across the cell cortex. Indeed, we report a reduction in cortical actin network tension elicited on secretagogue stimulation that is sensitive to myosin II inhibition. We reveal that the cortical actin network acts as a 'casting net' that undergoes activity-dependent relaxation, thereby driving tethered SVs towards the plasma membrane where they undergo Munc18-1-dependent docking.

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The control of cytoskeletal actin and exocytosis in intact and permeabilized adrenal chromaffin cells: role of calcium and protein kinase C

Cited by 101 publications

References 44 publications

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

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

Simultaneous measurements of cytosolic calcium and secretion in single bovine adrenal chromaffin cells by fluorescent imaging of fura-2 in cocultured cells.

Activity-driven relaxation of the cortical actomyosin II network synchronizes Munc18-1-dependent neurosecretory vesicle docking

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