scholarly journals Plasma membrane folds on the mast cell surface and their relationship to secretory activity.

1977 ◽  
Vol 74 (3) ◽  
pp. 690-697 ◽  
Author(s):  
S J Burwen ◽  
B H Satir
Keyword(s):  
Mast Cell ◽  
Plasma Membrane ◽  
Mast Cells ◽  
Cell Surface ◽  
Normal Population ◽  
Secretory Activity ◽  
Cell Type ◽  
Distinct Cell Type ◽  
Granule Membrane ◽  
Fold Formation

Changes in the surface morphology of secreting mast cells have been followed by scanning electron microscopy. Mast cells isolated from the rat peritoneal cavity have folds of plasma membrane that form snake-like ridges on their surfaces. Fold length varies considerably from cell to cell, whereas fold width and depth appear to remain relatively constant. To assess the possible relationship between secretory activity and surface folding, a seimquantitative method was used for measuring fold length in control and secreting populations. A positive correlation is found between secretion of histamine and the extent of membrane folds on the mast cell surface. The source of the membrane required for fold formation is probably secretory granule membrane incorporated into the plasma membranene as a result of exocytosis. Furthermore, a distinct cell type devoid of surface folds, designated as a raspberry-type cell, is found to occur as an integral part of a normal population of mast cells. This cell type is resistant to stimulation by polymyxin.

scholarly journals Dynamics of Plasma and Granule Membrane in Murine Bone Marrow-Derived Mast Cells after Re-stimulation

2015 ◽  
Vol 8 (1) ◽  
pp. 14-22
Author(s):  
Masahiro Kaneko ◽  
Arisa Yamada
Keyword(s):  
Bone Marrow ◽  
Mast Cell ◽  
Plasma Membrane ◽  
Mast Cells ◽  
Morphological Changes ◽  
Membrane Dynamics ◽  
Hematopoietic Stem ◽  
Murine Bone Marrow ◽  
Cell Functions ◽  
Granule Membrane

Mast cells are derived from hematopoietic stem cells and play important roles in allergic responses. Mast cells are long-lived compared with other granular cell types. Since the response of the individual mast cell after FcεRI-induced degranulation is unclear, the aim of this study was to analyze morphological changes in individual mast cells after restimulation. To observe plasma and granule membrane dynamics, AcGFP-actb (β-actin) and DsRed-monomer (DRM)- CD63 fusion constructs were introduced into bone marrow-derived mast cells (BMMCs). Furthermore, AcGFP-CD63 and DRM-Cma1 (mMCP-5) were introduced into BMMCs. Re-stimulation resulted in increased β-hexosaminidase release and cytokine mRNA expression similar to those observed during initial stimulation. Moreover, expression of FcεRI on BMMCs 24 h after initial stimulation was similar to that measured before initial stimulation. Changes in morphology of the plasma membrane and colocalization of granules and plasma membrane were observed after initial stimulation. BMMCs returned to normal 120 min after the initial stimulation. These phenomena were also observed in BMMCs after re-stimulation. BMMC chymase content decreased 20 min after stimulation but returned to near normal 24 h after stimulation. These findings suggest that mast cell functions can be maintained and that these cells can be repeatedly degranulated after FcεRI-mediated stimulation.

scholarly journals Anti-Allergic Drugs Tranilast and Ketotifen Dose-Dependently Exert Mast Cell-Stabilizing Properties

2016 ◽  
Vol 38 (1) ◽  
pp. 15-27 ◽  
Author(s):  
Asuka Baba ◽  
Masahiro Tachi ◽  
Yutaka Ejima ◽  
Yasuhiro Endo ◽  
Hiroaki Toyama ◽  
...  
Keyword(s):  
Mast Cell ◽  
Plasma Membrane ◽  
Mast Cells ◽  
Lucifer Yellow ◽  
Water Soluble ◽  
Confocal Imaging ◽  
Whole Cell ◽  
Peritoneal Mast Cells ◽  
Cell Recording ◽  
Cell Membrane Capacitance

Background: Anti-allergic drugs, such as tranilast and ketotifen, inhibit the release of chemokines from mast cells. However, we know little about their direct effects on the exocytotic process of mast cells. Since exocytosis in mast cells can be monitored electrophysiologically by changes in the whole-cell membrane capacitance (Cm), the absence of such changes by these drugs indicates their mast cell-stabilizing properties. Methods: Employing the standard patch-clamp whole-cell recording technique in rat peritoneal mast cells, we examined the effects of tranilast and ketotifen on the Cm during exocytosis. Using confocal imaging of a water-soluble fluorescent dye, lucifer yellow, we also examined their effects on the deformation of the plasma membrane. Results: Relatively lower concentrations of tranilast (100, 250 µM) and ketotifen (1, 10 µM) did not significantly affect the GTP-γ-S-induced increase in the Cm. However, higher concentrations of tranilast (500 µM, 1 mM) and ketotifen (50, 100 µM) almost totally suppressed the increase in the Cm, and washed out the trapping of the dye on the surface of the mast cells. Compared to tranilast, ketotifen required much lower doses to similarly inhibit the degranulation of mast cells or the increase in the Cm. Conclusions: This study provides electrophysiological evidence for the first time that tranilast and ketotifen dose-dependently inhibit the process of exocytosis, and that ketotifen is more potent than tranilast in stabilizing mast cells. The mast cell-stabilizing properties of these drugs may be attributed to their ability to counteract the plasma membrane deformation in degranulating mast cells.

scholarly journals Denatonium Benzoate Exposure Promotes IgE-Mediated Mast Cell Degranulation and Allergy By Upregulating FcεRIα Expression

2021 ◽  
Author(s):  
Huaping Xu ◽  
Xiaoyun Shi ◽  
Mengting Xie ◽  
Shiyu Xiao ◽  
Xin Li ◽  
...  
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Cell Surface ◽  
Surface Expression ◽  
Specific Ige ◽  
Mast Cell Degranulation ◽  
Cell Surface Expression ◽  
Denatonium Benzoate ◽  
Ige Mediated

Abstract Background: Denatonium benzoate (DB), one of the bitterest compounds known to man, is currently added to a wide range of products and is also used for alcohol denaturation. Some reports demonstrated that asthmatic symptoms are associated with DB exposure but the possible links between DB and IgE-mediated allergy susceptibility have not been examined to date. We investigated the effects of DB on IgE-mediated mast cell degranulation in vitro and in the ovalbumin (OVA)-induced mouse model of allergy.Methods: DB treatments were given to RBL-2H3 IgE-sensitized rat mast cell/basophil cells and KU812 human basophilic cells together with OVA-induced allergic BALB/c mice. Allergic mediator release, Ca2+ influx and OVA-specific IgE anaphylactic shock symptoms were measured along with the cell-surface expression of the α-subunit of high-affinity IgE receptor FcεRI on mast cells.Results: DB increases β-hexosaminidase (β-hex) release and Ca2+ mobilization in IgE-mediated activated RBL-2H3 and KU812 cells, and enhanced the cell-surface expression of FcεRIα. DB also promoted the severity of OVA-induced anaphylactic and diarrheic symptoms which was accompanied by mucus thickness in jejunum and the levels of β-hex, histamine and OVA-specific IgE in allergy mice, as well as the levels of FcεRIα mRNA and the FcεRIα proteinin isolated mucosal mast cells. Conclusions: DB treatments can promote the IgE-mediated mast cell degranulation in vitro and OVA-induced allergic susceptibility in mice by upregulating mast-cell-surface FcεR1α expression, providing evidence for DB exposure in promoting allergy susceptibility.

scholarly journals c-kit Gene was not transcribed in cultured mast cells of mast cell- deficient Wsh/Wsh mice that have a normal number of erythrocytes and a normal c-kit coding region

Blood ◽  
1992 ◽  
Vol 80 (6) ◽  
pp. 1448-1453 ◽  
Author(s):  
T Tono ◽  
T Tsujimura ◽  
U Koshimizu ◽  
T Kasugai ◽  
S Adachi ◽  
...  
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Germ Cells ◽  
Messenger Rna ◽  
Mutant Allele ◽  
Cell Type ◽  
Coding Region ◽  
Male And Female ◽  
Normal Number ◽  
Kit Gene

Abstract The Wsh is a mutant allele at the W (c-kit) locus of mice. Mice of Wsh/Wsh genotype have white hairs and black eyes. Although adult C57BL/6-Wsh/Wsh mice were not anemic, they showed a remarkable depletion of mast cells. Most homozygous or double heterozygous mutant mice at the W (c-kit) locus, of which mast-cell depletion was comparable to that of Wsh/Wsh mice, are deficient in germ cells. However, male and female Wsh/Wsh mice have an appreciable number of germ cells in their gonads. We investigated the mechanism of specific depletion of mast cells in Wsh/Wsh mice. Cultured mast cells (CMC) derived from the spleen of Wsh/Wsh mice neither attached to normal (+/+) fibroblasts nor survived in the coculture with +/+ fibroblasts. The c-kit messenger RNA (mRNA) was strongly expressed in +/+ CMC, but not detectable in Wsh/Wsh CMC. Despite the lack of c-kit mRNA in Wsh/Wsh CMC, the c-kit mRNA was normally detectable in the cerebellum and weakly detectable in the testis and spleen of Wsh/Wsh mice. No significant changes were found in the nucleotide sequence of the c-kit transcripts obtained from the cerebellum of Wsh/Wsh mice. Development of mast cells, erythrocytes, and germ cells in Wsh/Wsh mice appeared to be parallel with the magnitude of the c-kit gene expression in each cell type.

MAST cells in the dermis of rats repeatedly moistened with chloroform: An EM study

1994 ◽  
Vol 52 ◽  
pp. 242-243
Author(s):  
Y. Abu-Ghalyun ◽  
J. Shakhabneh ◽  
F. Al-Bagdadi
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Epithelial Cells ◽  
Blood Vessels ◽  
Toluidine Blue ◽  
Osmium Tetroxide ◽  
Free Cell ◽  
Lead Citrate ◽  
Cell Type ◽  
Uterine Glands

Mast cells are a free cell type, usually found in connective. The cells are common in the dermis and are abundant around blood vessels. Heterogeneity in the ultrastructure of mast cell has been documented. Variation of the morphology of the ultrastructure between the regional mast cells has been reported. Cellular relationships has been reported between mast cells and nerves, plasma cell, P neuropeptide and epithelial cells of uterine glands. The purpose of this study is to report the behaviour of mast cells in the dermis of rats repeatedly moistened with chloroform. Specimens were collected from the leg site of rats repeatedly moistened with chloroform, three times per day for three weeks. The collected specimens were immediately immersed in 2% glutaraldehyde in cacodylate sodium buffer at pH 7.4. Then the tissue was post fixed in 1% osmium tetroxide, dehydrated in ethyl alcohol, processed and embedded in Epon. One micron thick sections were stained with toluidine blue for orientation and silver sections were stained with lead citrate and urynal acetate.

Do Lipocalins Play a Role in Mast Cell Physiology?.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1645-1645
Author(s):  
Lennart E. Logdberg ◽  
Bo Akerstrom ◽  
Gregory A. Hair ◽  
Maria Allhorn ◽  
Tatyana Vikulina ◽  
...  
Keyword(s):  
Mast Cell ◽  
Monoclonal Antibodies ◽  
Mast Cells ◽  
Cell Surface ◽  
Gene Transcription ◽  
Cell Surface Receptor ◽  
Cell Surface Expression ◽  
Cell Physiology ◽  
Myeloma Protein ◽  
Surface Receptor

Abstract Introduction. Aiming to identify molecular properties of allergens responsible for their “intrinsic allergenicity”, we focus on a subset of xenogeneic lipocalins (LCs) that comprise the major mammalian respiratory allergens. These structurally and functionally homologous molecules likely possess conserved molecular motifs promoting IgE-dependent allergy. We hypothesize that such LC “allergenicity” depends on non-IgE interactions of LCs with components of the innate immune system. Herein we describe a possible basis for such interactions between LCs and mast cells. Materials and Methods. Two sources of human mast cells were used; primary cultures derived from peripheral blood CD34+ progenitor cells; or the LAD-2 cell line. Cells were cultured in serum-free medium with recombinant human stem cell factor (SCF; 100 ng/ml). Monoclonal antibodies to human gp330/megalin (MAb E11) were a kind gift of Prof. Lars Rask (Uppsala University, Sweden). Cell surface protein expression was assessed by flow cytometry and gene transcription was measured by real-time PCR. Results. Monoclonal antibodies to an endocytic cell surface receptor (megalin, also known as low-density lipoprotein receptor-related protein (LRP)-2) known to bind multiple LCs stained the mast cell lines. This putative expression of megalin by the mast cells corresponded to their transcription of megalin mRNA as shown as by PCR. Moreover, mast cell megalin gene transcription could be induced (≥ 1000-fold) by overnight culture with monomeric IgE myeloma protein (100 ng/ml), and such induction of the megalin message correlated with both an increase in cell surface expression of the molecule and the specific binding of a purified human LC (a-1 microglobulin). Conclusion. Megalin, an LC-binding cell surface receptor, appears to be constitutively expressed by both progenitor and mature mast cells, and its expression seems to be strongly upregulated by culture with monomeric IgE. This is consistent with a role for direct mast cell-LC interactions in the development of IgE-dependent allergy. In addition, and also of potential clinical relevance, endogenous LCs may play a functional role in normal mast cell physiology.

scholarly journals A Defect in Influenza A Virus Particle Assembly Specific to Primary Human Macrophages

mBio ◽  
2018 ◽  
Vol 9 (5) ◽  
Author(s):  
Sukhmani Bedi ◽  
Takeshi Noda ◽  
Yoshihiro Kawaoka ◽  
Akira Ono
Keyword(s):  
Plasma Membrane ◽  
Cell Surface ◽  
Influenza A ◽  
Cytochalasin D ◽  
Virus Release ◽  
Cell Type ◽  
Monocytic Cell ◽  
Monocytic Cell Line ◽  
Particle Assembly ◽  
Human Macrophages

ABSTRACTInfluenza A virus (IAV) propagates efficiently in epithelial cells, its primary target in the respiratory tract. In contrast, productive infection of most IAV strains is either blocked or highly inefficient in macrophages. The exact nature of the defect in IAV replication in human macrophages remains unknown. In this study, we showed that even compared to a monocytic cell line differentiated to macrophage-like cells, primary human monocyte-derived macrophages (MDM) are inefficient in IAV production, despite comparable levels of expression of viral glycoproteins at the plasma membrane. Correlative fluorescence scanning electron microscopy revealed that formation of budding structures at the cell surface is inefficient in MDM even though clustering of a viral glycoprotein, hemagglutinin (HA), is observed, suggesting that a step in IAV particle assembly is blocked in MDM. Using anin situproximity ligation assay, we further determined that HA associates with neuraminidase (NA) but fails to associate with another viral transmembrane protein, M2, at the MDM plasma membrane. Notably, the defects in HA-M2 association and particle assembly in MDM were reversed upon cytochalasin D treatment that inhibits actin polymerization. These results suggest that HA-M2 association on the plasma membrane is a discrete step in IAV production, which is susceptible to suppression by actin cytoskeleton in MDM. Virus release remained inefficient in MDM upon cytochalasin D treatment, suggesting the presence of an additional defect(s) in virus release in this cell type. Overall, our study revealed the presence of multiple cell-type-specific mechanisms negatively regulating IAV production at the plasma membrane in MDM.IMPORTANCEIdentification of host cell determinants promoting or suppressing replication of viruses has been aided by analyses of host cells that impose inherent blocks on viral replication. In this study, we show that primary human MDM, which are not permissive to IAV replication, fail to support virus particle formation. This defect is specific to primary human macrophages, since a human monocytic cell line differentiated to macrophage-like cells supports IAV particle formation. We further identified association between two viral transmembrane proteins, HA and M2, on the cell surface as a discrete assembly step, which is defective in MDM. Defective HA-M2 association and particle budding, but not virus release, in MDM are rescued by disruption of actin cytoskeleton, revealing a previously unknown, negative role for actin, which specifically targets an early step in the multistep IAV production. Overall, our study uncovered a host-mediated restriction of association between viral transmembrane components during IAV assembly.

Clarithromycin Dose-Dependently Stabilizes Rat Peritoneal Mast Cells

Chemotherapy ◽  
2016 ◽  
Vol 61 (6) ◽  
pp. 295-303 ◽  
Author(s):  
Itsuro Kazama ◽  
Kazutomo Saito ◽  
Asuka Baba ◽  
Tomohiro Mori ◽  
Nozomu Abe ◽  
...  
Keyword(s):  
Mast Cell ◽  
Plasma Membrane ◽  
Mast Cells ◽  
Water Soluble ◽  
Patch Clamp Technique ◽  
Membrane Deformation ◽  
Whole Cell Patch Clamp ◽  
Peritoneal Mast Cells ◽  
Rat Peritoneal Mast Cells ◽  
First Time

Background: Macrolides, such as clarithromycin, have antiallergic properties. Since exocytosis in mast cells is detected electrophysiologically via changes in membrane capacitance (Cm), the absence of such changes due to the drug indicates its mast cell-stabilizing effect. Methods: Employing the whole-cell patch clamp technique in rat peritoneal mast cells, we examined the effects of clarithromycin on Cm during exocytosis. Using a water-soluble fluorescent dye, we also examined its effect on deformation of the plasma membrane. Results: Clarithromycin (10 and 100 μM) significantly inhibited degranulation from mast cells and almost totally suppressed the GTP-γ-S-induced increase in Cm. It washed out the trapping of the dye on the surface of mast cells. Conclusions: This study provides for the first time electrophysiological evidence that clarithromycin dose-dependently inhibits the process of exocytosis. The mast cell-stabilizing action of clarithromycin may be attributable to its counteractive effect on plasma membrane deformation induced by exocytosis.

scholarly journals c-kit Gene was not transcribed in cultured mast cells of mast cell- deficient Wsh/Wsh mice that have a normal number of erythrocytes and a normal c-kit coding region

Blood ◽  
1992 ◽  
Vol 80 (6) ◽  
pp. 1448-1453
Author(s):  
T Tono ◽  
T Tsujimura ◽  
U Koshimizu ◽  
T Kasugai ◽  
S Adachi ◽  
...  
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Germ Cells ◽  
Messenger Rna ◽  
Mutant Allele ◽  
Cell Type ◽  
Coding Region ◽  
Male And Female ◽  
Normal Number ◽  
Kit Gene

The Wsh is a mutant allele at the W (c-kit) locus of mice. Mice of Wsh/Wsh genotype have white hairs and black eyes. Although adult C57BL/6-Wsh/Wsh mice were not anemic, they showed a remarkable depletion of mast cells. Most homozygous or double heterozygous mutant mice at the W (c-kit) locus, of which mast-cell depletion was comparable to that of Wsh/Wsh mice, are deficient in germ cells. However, male and female Wsh/Wsh mice have an appreciable number of germ cells in their gonads. We investigated the mechanism of specific depletion of mast cells in Wsh/Wsh mice. Cultured mast cells (CMC) derived from the spleen of Wsh/Wsh mice neither attached to normal (+/+) fibroblasts nor survived in the coculture with +/+ fibroblasts. The c-kit messenger RNA (mRNA) was strongly expressed in +/+ CMC, but not detectable in Wsh/Wsh CMC. Despite the lack of c-kit mRNA in Wsh/Wsh CMC, the c-kit mRNA was normally detectable in the cerebellum and weakly detectable in the testis and spleen of Wsh/Wsh mice. No significant changes were found in the nucleotide sequence of the c-kit transcripts obtained from the cerebellum of Wsh/Wsh mice. Development of mast cells, erythrocytes, and germ cells in Wsh/Wsh mice appeared to be parallel with the magnitude of the c-kit gene expression in each cell type.

ATP is required in platelet serotonin exocytosis for protein phosphorylation and priming of secretory vesicles docked on the plasma membrane

1996 ◽  
Vol 109 (1) ◽  
pp. 113-118
Author(s):  
T. Morimoto ◽  
S. Ogihara
Keyword(s):  
Plasma Membrane ◽  
Protein Phosphatase ◽  
Secretory Activity ◽  
Electron Microscopic ◽  
Dense Granules ◽  
Granule Membrane ◽  
Microscopic Studies ◽  
Amorphous Structures ◽  
Gamma S

Calcium-evoked secretion generally requires the presence of millimolar concentrations of Mg-ATP. We investigated the role of Mg-ATP in the secretion of serotonin from electropermeabilized bovine platelets. The secretion of serotonin was lost within 5 minutes when the Mg-ATP concentration was diluted to less than 0.1 mM, but was maintained when ATP-gamma S (adenosine 5′-O-3-thiotriphosphate) was used instead of ATP. Okadaic acid, a potent inhibitor of protein phosphatase, could also maintain the exocytotic activity even when ATP was diluted. Decrease in the secretory activity was paralleled by a decrease in phosphorylation level of four proteins after dilution of ATP, but the activity was maintained when the thiophosphorylation level of these proteins was maintained. Two of these proteins were digested by a protease, calpain, which has been shown to lead to a loss in the exocytotic activity. Electron microscopic studies showed that calcium did not induce the formation of distinct bridge-like structures between the granule membrane and the plasma membrane in Mg-ATP-diluted cells, previously shown as the structure transiently formed prior to fusion of the two membranes. Anchorage of the secretory dense granules to the plasma membrane and the presence of the amorphous structures between the granules and the plasma membrane were unchanged by dilution of ATP. These results indicate that ATP is not required for the anchorage itself, but is required to prime anchored granules for calcium-triggered secretion. Maintenance of the phosphorylated state of proteins by ATP enables the calcium trigger to form the bridge-like structures preceding membrane fusion events.

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