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 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

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 Anti-immunoglobulin-induced histamine secretion by rat peritoneal mast cells studied by immunoferritin electron microscopy.

1975 ◽  
Vol 142 (2) ◽  
pp. 391-402 ◽  
Author(s):  
D Lawson ◽  
C Fewtrell ◽  
B Gomperts ◽  
M Raff
Keyword(s):  
Electron Microscopy ◽  
Mast Cell ◽  
Mast Cells ◽  
Histamine Release ◽  
Histamine Secretion ◽  
Calcium Dependent ◽  
Peritoneal Mast Cells ◽  
Rat Peritoneal Mast Cells

We have used ferritin-conjugated divalent and monovalent anti-Ig antibodies to study simultaneously, histamine secretion and the ultrastructural distribution and redistribution of Ig receptors on rat peritoneal mast cells. We conclude that (a) divalent anti-Ig is required for both receptor redistribution and for calcium-dependent degranulation and histamine release, (b) divalent anti-Ig induces patching and pinocytosis but not capping of Ig molecules, (c) neither capping nor pinocytosis are required for triggering and if clustering is necessary, then less than 10 Ig molecules are required per cluster, and (d) degranulation (and histamine release) is not an all or none response of the mast cell.

scholarly journals MECHANISMS OF IMMUNOLOGIC INJURY OF RAT PERITONEAL MAST CELLS

1966 ◽  
Vol 124 (3) ◽  
pp. 379-395 ◽  
Author(s):  
Elmer L. Becker ◽  
K. Frank Austen
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Guinea Pig ◽  
Histamine Release ◽  
Response Curve ◽  
Peritoneal Mast Cells ◽  
Rat Peritoneal Mast Cells ◽  
Logarithmic Relationship ◽  
Antigen Antibody ◽  
The Complement System

The ability of a number of p-nitrophenylethyl, alkyl phenylalkyl, chloroalkyl, and aminoalkyl phosphonates to inhibit the homocytotropic antibody-mediated release of histamine from rat peritoneal mast cells has been tested. The effectiveness of these same phosphonates against the activated first component of rat complement (C'1a) has also been investigated. The rat mast cell esterase activated by the reaction of antigen and homocytotropic antibody resembles chymotrypsin in its reactivity with the phenylalkyl and chloroalkyl phosphonate, but is unlike this protease in its greater responsiveness to the 5-aminopentyl phosphonate relative to the pentyl phosphonate. The antigen-homocytotropic antibody-activated mast cell esterase and chymotrypsin, thus, appear to be similar, but different enzymes; i.e., they are parazymes (see reference 4, p. 501). There are distinct differences in the pattern of inhibition given by the phenylalkyl and aminoalkyl and alkyl phosphonates of the homocytotropic antibody-mediated histamine release from rat peritoneal mast cells and from guinea pig lung slices. On the basis of these differences it is concluded that the esterases activated by the combination of antigen and homocytotropic antibody on the mast cells of the two species are not the same. The arithmetic dose response curve found for the action of the phosphonates on the antigen-induced histamine release from rat peritoneal mast cells contrasted sharply with the logarithmic relationship found when these same inhibitors acted on the guinea pig lung system. This suggests that in addition to the antigen-antibody-activated esterases being unlike, the detailed mode of histamine release from the mast cells of the guinea pig lung differs from that of the mast cells of the rat peritoneum. Distinct and large differences were found in the pattern of inhibition of histamine release from rat peritoneal mast cells and of rat C'1a given by the phenylalkyl, and chloroalkyl and alkyl phosphonates implying that esterase activated by the combination of antigen with the sensitized rat peritoneal mast cells is not C'1a. Thus, the results with the peritoneal mast cells lead to the same conclusion as the previous work with guinea pig lung slices; i.e., the antigen-antibody-activated esterase involved in the homocytotropic antibody-mediated release of histamine is not part of the complement system.

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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