Control of exocytosis in adrenal chromaffin cells

Burgoyne, Robert D.

doi:10.1016/0304-4157(91)90024-q

Cited by 248 publications

(183 citation statements)

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“…In chromaffin cells, disassembly of the cytoskeleton seemed to be needed for the secretory granules to move to the plasma membrane. Gelsolin or scinderin, which are calcium-dependent actin filament severing proteins, may disrupt actin networks (7,8). Un like cytoskeletal assembly, there was no lag time in hista mine release induced by DSA (20 pg/ml) and compound 48/80 (5 pg/ml) in Figs.…”

Section: Discussionmentioning

confidence: 85%

An Initial Signal of Activation of Rat Peritoneal Mast Cells Stimulated by Datura stramonium Agglutinin: A Confocal Fluorescence Microscopic Analysis of Intracellular Calcium Ion and Cytoskeletal Assembly

Suzuki-Nishimura

Furuno

Uchida

et al. 1994

Japanese Journal of Pharmacology

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-A confocal fluorescence microscope using fluo-3 and 9-(dicyanovinyl)-julolidine (DCVJ) was used to study the mast cell activation by the N-acetyl glucosamine oligomer specific lectin Datura stramonium agglutinin (DSA) and inhibition by antagonist lectins having affinity to N-acetyl glucosamine G1cNAc oligomer specific lectin DSA is a useful tool for examining the pertussis toxin-sensitive mechanisms of histamine release from rat peritoneal mast cells, as previ ously described (1). The G1cNAc-specific lectins WGA and STA were antagonists of DSA and compound 48/80. They may occupy sugar residues of glycoproteins that serve as binding sites for DSA, which overlap one of the binding sites of compound 48/80, prevent access of DSA and compound 48/80 to the binding sites and inhibit the histamine release induced by DSA and compound 48/80 (1-3). Con A, however, did not interfere with the inter actions and did not inhibit the histamine release, since it recognizes high-mannose sugar-residues but not G1cNAc oligomer (1, 3).DSA releases histamine (1), but it is not clear whether the increase in [Ca"]; is one of the initial signals trigger ing histamine release or whether the histamine releasing mechanisms are coupled with changing cytoskeletal assem bly. Confocal fluorescence microscopy, a new technol ogy, revealed important information about cell activation. Real time changes in the fluorescence of indicator dyes and fluorescent molecular rotors in living cells can be measured. In this study, we used the calcium ion indicator fluo-3 and the fluorescent molecular rotor DCVJ. Fluo-3 is a useful dye, but intracellular calcium ion concentra tion can not be calculated from its fluorescence. The inten sity of DCVJ fluorescence increased with the increasing cytoskeletal assembly of actin and tubulin molecules, and cytochalasin D inhibited the increase in the DCVJ fluores cence intensity in activated mast cells (4). We also exam ined the mechanisms of action of antagonist lectins (1) using real time measurement. To clarify the mechanisms of the DSA-induced histamine release, confocal fluores cence microscopic imaging using Con A and the antago nist lectins WGA and STA was carried out.

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Section: Discussionmentioning

confidence: 85%

An Initial Signal of Activation of Rat Peritoneal Mast Cells Stimulated by Datura stramonium Agglutinin: A Confocal Fluorescence Microscopic Analysis of Intracellular Calcium Ion and Cytoskeletal Assembly

Suzuki-Nishimura

Furuno

Uchida

et al. 1994

Japanese Journal of Pharmacology

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“…The release due to GppNHp was 2.5 _+ 0.2% of total cellular catecholamine above basal from 17 cell batches. This extent of release could conceivably be accounted for by release of only docked secretory granules (corresponding to 1.5-3% of total granules per cell [15,39]). This is potentially of importance since it has been suggested that SNARE proteins of docked vesicles would not be accessible to neurotoxins [38].…”

Section: Resultsmentioning

confidence: 99%

“…Inhibition of exocytosis in a non-neuronal cell by these neurotoxins was first shown for adrenal chromaffin cells [14] which have been an important model cell for the study of CaZ+-regulated exocytosis [15,16].…”

Section: Introductionmentioning

confidence: 99%

“…These data on insulin-secreting cells, coupled with studies on constitutive vesicle traffic in epithelial cells in which basolateral but not apical exocytosis is inhibited by botulinum neurotoxins [32], appear to suggest that exocytosis could occur by either SNARE-dependent or SNARE-independent pathways. Exocytosis in chromaffin cells is triggered by Ca 2+ [15,16] but exocytosis can also be activated in a CaZ+-independent manner by GTP analogues in permeabilised chromaffin cells [33][34][35][36]. We have, therefore, used botulinum neurotoxins to investigate whether exocytosis activated by a non-hydrolysable GTP analogue in permeabilised chromaffin cells, occurs by a SNARE-dependent mechanism based on sensitivity to recombinant neurotoxin light chains.…”

Section: Introductionmentioning

confidence: 99%

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Botulinum neurotoxin light chains inhibit both Ca²⁺‐induced and GTP analogue‐induced catecholamine release from permeabilised adrenal chromaffin cells

Glenn

Burgoyne

1996

FEBS Letters

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Using digitonin-permeabilised bovine adrenal chromaffin cells, the effects of botulinum neurotoxin light chains on exocytosis triggered by Ca 2+ or by GppNHp were examined. Botulinum neurotoxin D light chain, prepared as a His6-tagged recombinant protein, cleaved VAMP and substantially inhibited catecholamine release due to Ca 2+ and GppNHp. Botulinum neurotoxin C1 and E light chains produced partial inhibition of both Ca 2+-and GppNHp-induced catecholamine release. These results suggest that Ca2+-dependent exocytosis and Ca 2+-independent exocytosis triggered by a noa-hydrolysable GTP analogue occurs via a SNARE-dependent mechanism in chromaffin cells.

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“…Though a change in the degree of phosphorylation of different proteins is a key event accompanying triggered secretory activity in many cells, no phosphoprotein (PP) has so far been shown to be directly involved in the regulation of vesicle-cell-membrane interaction for fusion (for reviews, see Hemmings et al, 1989;Plattner, 1989;Burgoyne, 1991). Some of the uncertainties come from superposition of different steps, such as organelle docking, membrane fusion, resealing and internalization.…”

Section: Introduction Materials and Methodsmentioning

confidence: 99%

A cortical phosphoprotein (‘PP63’) sensitive to exocytosis triggering in Paramecium cells. Immunolocalization and quenched-flow correlation of time course of dephosphorylation with membrane fusion

Höhne-Zell

Knoll

Riedel-Gras

et al. 1992

Biochemical Journal

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We had previously shown that a phosphoprotein of 63 kDa ('PP63') is rapidly and selectively dephosphorylated during synchronous (< 1 s) trichocyst exocytosis in Paramecium cells and then rephosphorylated within < 1 min [Zieseniss & Plattner (1985) J. Cell Biol. 101, 2028-2035. Using a new quenched-flow device, we now find a strict correlation between PP63 dephosphorylation and the process of membrane fusion, both occurring within 80 ms. Uptake of 32P over 90 min, followed by exocytosis and rephosphorylation for 1 min, results in a rather selective phosphorylation of the dephosphorylated form, P63, to PP63. Solubilization by repeated freezing and thawing allows isolations of P63 and PP63. On isoelectric focusing autoradiograms they have pl values of 6.05, 5.95 (major spots), 5.85 and 5.75. All spots are sensitive to alkaline, but not to acidic, hydrolysis (except for the pl-6.05 spot). On two-dimensional-gel autoradiograms the most prominent spot, of pl 5.95, is most extensively de-and re-phosphorylated. This spot, from de-and rephosphorylated samples, was used to produce monospecific antibodies. A cortical localization of PP63 was revealed by producing Western blots from isolated cell-surface fragments ('cortices') and by immunofluorescence labelling. We assume that both P63 and PP63 are attached to cortical structures, e.g. around trichocysts, though they are partly soluble. This localization and the strict correlation of PP63 dephosphorylation with exocytotic membrane fusion suggests a role in fusion regulation. INTRODUCTION MATERIALS AND METHODSThough a change in the degree of phosphorylation of different proteins is a key event accompanying triggered secretory activity in many cells, no phosphoprotein (PP) has so far been shown to be directly involved in the regulation of vesicle-cell-membrane interaction for fusion (for reviews, see Hemmings et al., 1989;Plattner, 1989;Burgoyne, 1991). Some of the uncertainties come from superposition of different steps, such as organelle docking, membrane fusion, resealing and internalization. A synchronous system allowing for the selective analysis of specific steps would be of evident advantage. This is the case with Paramecium cells, whose trichocysts can be released synchronously upon triggering with certain polyaminated secretagogues . The recent development of a quenched-flow procedure applicable to such fragile cells now allows for simultaneous analysis by ultrastructural and biochemical methods, specifically of the membrane fusion process. The present investigation was focused on the dephosphorylation of a PP of 63 kDa (PP63, formerly 65 kDa), a salient feature of trichocyst exocytosis (Gilligan & Satir, 1982;Zieseniss & Plattner, 1985). We analysed the precise time course of PP63 dephosphorylation, the occurrence of PP63 isoforms of different pl values and their sensitivity to exocytosis triggering, as well as the subcellular localization of PP63 and its dephosphorylated form (P63). We prepared monospecific antibodies for Western blots and for immunocytochemical ...

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Control of exocytosis in adrenal chromaffin cells

Cited by 248 publications

References 258 publications

An Initial Signal of Activation of Rat Peritoneal Mast Cells Stimulated by Datura stramonium Agglutinin: A Confocal Fluorescence Microscopic Analysis of Intracellular Calcium Ion and Cytoskeletal Assembly

An Initial Signal of Activation of Rat Peritoneal Mast Cells Stimulated by Datura stramonium Agglutinin: A Confocal Fluorescence Microscopic Analysis of Intracellular Calcium Ion and Cytoskeletal Assembly

Botulinum neurotoxin light chains inhibit both Ca²⁺‐induced and GTP analogue‐induced catecholamine release from permeabilised adrenal chromaffin cells

A cortical phosphoprotein (‘PP63’) sensitive to exocytosis triggering in Paramecium cells. Immunolocalization and quenched-flow correlation of time course of dephosphorylation with membrane fusion

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Control of exocytosis in adrenal chromaffin cells

Cited by 248 publications

References 258 publications

An Initial Signal of Activation of Rat Peritoneal Mast Cells Stimulated by Datura stramonium Agglutinin: A Confocal Fluorescence Microscopic Analysis of Intracellular Calcium Ion and Cytoskeletal Assembly

An Initial Signal of Activation of Rat Peritoneal Mast Cells Stimulated by Datura stramonium Agglutinin: A Confocal Fluorescence Microscopic Analysis of Intracellular Calcium Ion and Cytoskeletal Assembly

Botulinum neurotoxin light chains inhibit both Ca2+‐induced and GTP analogue‐induced catecholamine release from permeabilised adrenal chromaffin cells

A cortical phosphoprotein (‘PP63’) sensitive to exocytosis triggering in Paramecium cells. Immunolocalization and quenched-flow correlation of time course of dephosphorylation with membrane fusion

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Botulinum neurotoxin light chains inhibit both Ca²⁺‐induced and GTP analogue‐induced catecholamine release from permeabilised adrenal chromaffin cells