Exocytosis (Secretory Granule Extrusion) Induced by Injection of Calcium into Mast Cells

1973 ◽  
Vol 51 (12) ◽  
pp. 1001-1004 ◽  
Author(s):  
T. Kanno ◽  
D. E. Cochrane ◽  
W. W. Douglas
Keyword(s):  
Mast Cells ◽  
Secretory Granule ◽  
Secretory Granules ◽  
Intracellular Concentration ◽  
General Requirement ◽  
Peritoneal Mast Cells ◽  
Direct Support ◽  
Ca Ions ◽  
Stimulus Secretion Coupling ◽  
Rat Peritoneal Mast Cells

Injection of Ca (but not Mg or K) through micropipettes placed within rat peritoneal mast cells elicited extrusion of secretory granules. The result is considered direct support for the view that a rise in the intracellular concentration of Ca ions suffices to induce exocytosis and accounts for the general requirement for Ca in stimulus–secretion coupling.

scholarly journals Immunocytochemical localization of histamine in secretory granules of rat peritoneal mast cells with conventional or rapid microwave fixation and an ultrastructural post-embedding immunogold technique.

1992 ◽  
Vol 40 (9) ◽  
pp. 1247-1256 ◽  
Author(s):  
G R Login ◽  
S J Galli ◽  
A M Dvorak
Keyword(s):  
Electron Microscopy ◽  
Mast Cells ◽  
Guinea Pig ◽  
Secretory Granules ◽  
Fixation Method ◽  
Structural Localization ◽  
Thin Sections ◽  
Aldehyde Fixation ◽  
Peritoneal Mast Cells ◽  
Rat Peritoneal Mast Cells

We used a post-embedding immunogold labeling approach to define the fine-structural localization of histamine in rat peritoneal mast cells that were fixed using either standard aldehyde fixation or a fast microwave-aldehyde fixation method. Specimens were processed routinely for electron microscopy. Thin sections were exposed first to guinea pig antihistamine antiserum and then to gold-conjugated goat IgG directed against guinea pig IgG. By transmission electron microscopy, gold particles were localized to the matrix of cytoplasmic granules. Control sections treated with non-immune sera did not show labeling of mast cells. Adsorption of antihistamine antiserum with purified histamine or histamine bound to agarose showed a significant reduction (p less than 0.005) in granule staining. We also confirmed that our isolation procedures yielded functionally competent mast cells which released histamine when stimulated with sheep anti-rat IgE antiserum or with compound 48/80. These studies define the conditions of fixation for electron microscopy that are appropriate for the localization of histamine in the granule matrix of rat peritoneal mast cells.

scholarly journals Evidence for a role of phosphatidylinositol turnover in stimulus–secretion coupling. Studies with rat peritoneal mast cells

1979 ◽  
Vol 178 (3) ◽  
pp. 681-687 ◽  
Author(s):  
Shamshad Cockcroft ◽  
Bastien D. Gomperts
Keyword(s):  
Mast Cells ◽  
Concanavalin A ◽  
Immunoglobulin E ◽  
Histamine Secretion ◽  
Phosphatidylinositol Turnover ◽  
Phosphatidylinositol Response ◽  
Peritoneal Mast Cells ◽  
Stimulus Secretion Coupling ◽  
Rat Peritoneal Mast Cells

Histamine secretion and phosphatidylinositol turnover were compared in antigen-sensitized rat peritoneal mast cells stimulated with a number of different ligands. A small and variable increase in the incorporation of [32P]Pi and of [3H]inositol into phosphatidylinositol was observed when the cells were treated with immunoglobulin E-directed ligands (antigens and concanavalin A), and this was accompanied by a low amount of secretion (<10% of total cell histamine). In the presence of added phosphatidylserine, the addition of immunoglobulin E-directed ligands invariably led to an enhanced rate (approx. 4-fold) of labelling of phosphatidylinositol and, in the presence of Ca2+, this was accompanied by the secretion of histamine. The labelling of phosphatidylinositol and histamine secretion were also stimulated by chymotrypsin and compound 48/80. Whereas the phosphatidylinositol response did not require the presence of extracellular Ca2+, the secretion of histamine was either enhanced or dependent on extracellular Ca2+ (depending on the ligand used). The dependence on ligand concentration for the phosphatidylinositol response and histamine secretion were similar. The increased isotopic incorporation into phosphatidylinositol continued for about 1h although histamine secretion (elicited with concanavalin A) stopped within 2min. These results support the proposition that metabolic events involving phosphatidylinositol play a necessary intermediate role in the regulation of Ca2+ channels by ligand-activated receptors.

PT pretreatment inhibits 48/80-induced activation of Ca(+)-permeable channels in rat peritoneal mast cells

1990 ◽  
Vol 259 (5) ◽  
pp. C715-C722 ◽  
Author(s):  
M. Kuno ◽  
J. Kawaguchi ◽  
M. Mukai ◽  
F. Nakamura
Keyword(s):  
Mast Cells ◽  
Patch Clamp ◽  
Room Temperature ◽  
Plasma Membranes ◽  
Patch Clamp Technique ◽  
Permeable Channel ◽  
Peritoneal Mast Cells ◽  
Fluorescence Change ◽  
Stimulus Secretion Coupling ◽  
Rat Peritoneal Mast Cells

We recently reported that the secretagogue, compound 48/80, activated Ca2(+)-permeable channels of mast cells possibly via a second messenger [Kuno, Okada, and Shibata. Am. J. Physiol. 256 (Cell Physiol. 25): C560-C568, 1989]. The effects of pretreatment with pertussis toxin (PT) on compound 48/80 (48/80)-induced activation of the Ca2(+)-permeable channel have now been investigated by measuring Ca2+ signals of cell suspensions using the Ca2+ indicator fura-2 and by recording Ba2+ currents through the channel using the patch-clamp technique. In the presence of extracellular Ca2+, the fluorescence change was biphasic, with an immediate rise and a delayed peak at room temperature. The delayed peak, mainly due to Ca2+ entry through plasma membranes, was greatly reduced by pretreatment with PT. The quenching of the fluorescence by 48/80-induced Mn2+ influx was also decreased by PT, whereas the Ca2+ transients due to Ca2+ release from the intracellular stores apparently did not change. In patch-clamp recordings from cell-attached patches with pipettes containing isotonic Ba2+, the 48/80-induced Ba2+ currents were either suppressed or delayed in the PT-treated cells, under conditions where degranulation was absent. These results suggest that PT-sensitive GTP-binding protein is involved in activating the Ca2(+)-permeable channel in mast cells during stimulus-secretion coupling.

scholarly journals Latexin, a carboxypeptidase A inhibitor, is expressed in rat peritoneal mast cells and is associated with granular structures distinct from secretory granules and lysosomes

2000 ◽  
Vol 346 (3) ◽  
pp. 817 ◽  
Author(s):  
Yoshihiko URATANI ◽  
Keiko TAKIGUCHI-HAYASHI ◽  
Nobuhiko MIYASAKA ◽  
Michio SATO ◽  
Ming-hao JIN ◽  
...  
Keyword(s):  
Mast Cells ◽  
Secretory Granules ◽  
Carboxypeptidase A ◽  
Peritoneal Mast Cells ◽  
Rat Peritoneal Mast Cells

Inhibitors of the arachidonic acid cascade dissociate 48/80-induced Ca2+ influx and Ca2+ release in mast cells

1993 ◽  
Vol 264 (4) ◽  
pp. C912-C917 ◽  
Author(s):  
M. Kuno ◽  
J. Kawawaki ◽  
T. Shibata ◽  
H. Gotani
Keyword(s):  
Arachidonic Acid ◽  
Mast Cells ◽  
Nordihydroguaiaretic Acid ◽  
Arachidonic Acid Cascade ◽  
Lipoxygenase Inhibitor ◽  
Low Concentrations ◽  
Peritoneal Mast Cells ◽  
Stimulus Secretion Coupling ◽  
Rat Peritoneal Mast Cells ◽  
Full Activation

Effects of inhibitors of the arachidonic acid cascade on Ca2+ release from intracellular stores and Ca2+ influx through the plasma membrane during stimulus-secretion coupling were examined using rat peritoneal mast cells loaded with fura-2. Compound 48/80 (48/80) was used as a secretagogue. A phospholipase inhibitor, p-bromophenacyl bromide (PBPB), or a lipoxygenase inhibitor, nordihydroguaiaretic acid (NDGA), inhibited the 48/80 (1 microgram/ml)-induced release of histamine, Ca2+, and Mn2+ influxes, but the cyclooxygenase inhibitor, indomethacin (approximately 50 microM), inhibited neither Ca2+ nor Mn2+ influxes. The Ca2+ release induced by 1 microgram/ml of 48/80 was little inhibited by PBPB, NDGA, or indomethacin. The Ca2+ release was activated and saturated with lower concentrations of 48/80 than was the Ca2+ influx. The percent inhibition of the Ca2+ release by 25 microM PBPB was increased by lowering the concentration of 48/80, but NDGA (10 microM) did not inhibit the Ca2+ release induced by low concentrations of 48/80 (0.03-0.1 microgram/ml). These results suggest that activation of the Ca2+ release and the Ca2+ influx were differently regulated and that full activation of Ca2+ influx needs the arachidonic acid cascade produced by higher concentrations of 48/80 than does the Ca2+ release. Lipoxygenase metabolites of arachidonic acid are potential modulators of the Ca2+ influx.

Washout phenomena in dialyzed mast cells allow discrimination of different steps in stimulus-secretion coupling

1987 ◽  
Vol 7 (4) ◽  
pp. 313-321 ◽  
Author(s):  
Reinhold Penner ◽  
Michael Pusch ◽  
Erwin Neher
Keyword(s):  
Mast Cells ◽  
Calcium Transients ◽  
Capacitance Measurement ◽  
Patch Clamp Technique ◽  
Peritoneal Mast Cells ◽  
Cell Recording ◽  
Stimulus Secretion Coupling ◽  
Rat Peritoneal Mast Cells ◽  
Indicator Dye ◽  
Signal Mediators

Transient increases of intracellular calcium and exocytotic activity of rat peritoneal mast cells following stimulation with compound 48/80 were monitored using the Ca-indicator dye fura-2 and the capacitance measurement technique. It is known that mast cells very rapidly lose their secretory response towards antigenic or compound 48/80-induced stimulation in the whole-cell recording configuration of the patch-clamp technique due to “washout” of signal mediators. In contrast, we found that calcium transients remained unaffected by intracellular dialysis for as long as 10 min. The fast “washout” phenomenon of exocytosis could be overcome by supplementing the pipette filling solution with guanosinetriphosphate (GTP) indicating a major role for GTP-binding proteins in secretion. The restoration of exocytosis was transient and decayed within three minutes, suggesting diffusional escape of one or several other cytoplasmic substances involved in stimulus-secretion coupling. Quantitative aspects of this process and the implications of its differential effects on Ca-transients versus secretion are discussed.

scholarly journals Immunocytochemical Localization of Heparin in Secretory Granules of Rat Peritoneal Mast Cells Using a Monoclonal Anti-heparin Antibody (ST-1)1

1997 ◽  
Vol 45 (2) ◽  
pp. 231-235 ◽  
Author(s):  
Sonia M. Oliani ◽  
Edna Freymüller ◽  
Helio K. Takahashi ◽  
Anita H. Straus
Keyword(s):  
Mast Cells ◽  
Intestinal Mucosa ◽  
Sodium Periodate ◽  
Secretory Granules ◽  
Protein A ◽  
Immunogold Labeling ◽  
Transmission Electron ◽  
Peritoneal Mast Cells ◽  
Ultrathin Sections ◽  
Rat Peritoneal Mast Cells

We performed immunogold labeling with an ST-1 monoclonal antibody (IgM), specific for intact heparin, to define the subcellular localization of heparin in mast cells. Rat peritoneal mast cells were fixed by a modified Karnovsky method and embedded in Araldite. Ultrathin sections were first treated with sodium periodate and then sequentially incubated with MAb ST-1, rabbit anti-mouse IgM, and protein A-gold. By transmission electron microscopy, gold particles were localized inside cytoplasmic granules of peritoneal mast cells. In contrast, with the same procedure, no labeling was observed in mast cells from rat intestinal mucosa. Control sections of rat peritoneal or intestinal mucosa mast cells treated with an irrelevant MAb (IgM) did not show any labeling. Treatment with nitrous acid abolished the reactivity of MAb ST-1 with peritoneal mast cells. These results show that different mast cells can be identified regarding their heparin content by immunochemical procedures using MAb ST-1.

scholarly journals Latexin, a carboxypeptidase A inhibitor, is expressed in rat peritoneal mast cells and is associated with granular structures distinct from secretory granules and lysosomes

2000 ◽  
Vol 346 (3) ◽  
pp. 817-826 ◽  
Author(s):  
Yoshihiko URATANI ◽  
Keiko TAKIGUCHI-HAYASHI ◽  
Nobuhiko MIYASAKA ◽  
Michio SATO ◽  
Ming-hao JIN ◽  
...  
Keyword(s):  
Mast Cells ◽  
Secretory Granules ◽  
Minority Population ◽  
Carboxypeptidase A ◽  
Unique Type ◽  
Peritoneal Mast Cells ◽  
Neural Tissues ◽  
Rat Peritoneal Mast Cells ◽  
Silica Gel Particles ◽  
Rat Mast Cells

Latexin, a protein possessing inhibitory activity against rat carboxypeptidase A1 (CPA1) and CPA2, is expressed in a neuronal subset in the cerebral cortex and cells in other neural and non-neural tissues of rat. Although latexin also inhibits mast-cell CPA (MCCPA), the expression of latexin in rat mast cells has not previously been confirmed. In the present study we examined the expression and subcellular localization of latexin in rat peritoneal mast cells. Western blot and reverse-transcriptase-mediated PCR analyses showed that latexin was contained and expressed in the rat peritoneal mast cells. Immunocytochemically, latexin immunofluorescence was localized on granular structures distinct from MCCPA-, histamine- or cathepsin D-immunopositive granules. Immunoelectron microscopy revealed that latexin was associated with a minority population of granules. The latexin-associated granules were separated from MCCPA- or histamine-containing granules on a self-generating density gradient of polyvinylpyrrolidone-coated silica-gel particles (Percoll). Treatments with high ionic strength and heparinase released latexin from the granules, suggesting that latexin is non-covalently associated with a heparin-like component of the granules. MCCPA and histamine were released from the mast cells after non-immunological and immunological stimulation with compound 48/80, A23187 and anti-IgE antibody, whereas latexin was not released. These results show that latexin is synthesized in rat peritoneal mast cells and suggest that it is associated with a unique type of intracellular granules distinct from MCCPA- and histamine-containing secretory granules and lysosomes.

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