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  • 1
    Electronic Resource
    Electronic Resource
    Fasting or dexamethasone treatment reduce protease content in rat lung mast cells and modulation of histamine synthesis by H3 receptors (1994)
    Springer
    Inflammation research 42 (1994), S. 7-12 
    Details
    ISSN: 1420-908X
    Keywords: Rat mast cell proteases ; Histamine presynaptic receptor ; Dexamethasone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The sensitivity of mast cells to H3-receptor modulation was studied in rat lung under various hormonal conditions. The heterogeneity of mast cell subpopulations in rat lung was assessed by the tissue content of rat mast cell protease I (RMCP I) and rat mast cell protease II (RMCP II). After 24 h fasting, concentrations of RMCP I were unchanged whereas the concentration of RMCP II was significantly reduced by 49%. The [3H]histamine (HA) synthesis was concomitantly decreased by 35%. In addition, the modulation of [3H]HA, synthesis by the H3 receptor agonist, (R)α-methylHA and by the antagonist, thioperamide, observed in control rats, was lost in fasted rats. Single and repeated, administrations of dexamethasone did not influence RMCPI concentrations, but decreased the concentrations of RMCP II with a parallel decrease in [3H]HA synthesis. The inhibitory effect of (R)α-methylHA on [3H]HA synthesis was also reduced. These results suggest that a subpopulation of RMCP II-containing mast cells, very sensitive to environmental factors, could be the mast cells synthesizing HA in an H3-receptor-dependant manner.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02014292
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  • 2
    Electronic Resource
    Electronic Resource
    Constitutive secretion of the granule chymase mouse mast cell protease-1 and the chemokine, CCL2, by mucosal mast cell homologues (2003)
    Oxford, UK : Blackwell Science Ltd
    Clinical & experimental allergy 33 (2003), S. 0 
    Details
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background The mucosal mast cell (MMC) granule-specific β-chymase, mouse mast cell protease-1 (mMCP-1), is released systemically into the bloodstream early in nematode infection before parasite-specific IgE responses develop and TGF-β1 induces constitutive release of mMCP-1 by homologues of MMC in vitro. Intraepithelial MMC may also express the chemokine CCL2 (monocyte chemotactic protein-1) during nematode infection but the expression of this chemokine by MMC homologues has not been investigated.Objective To investigate the expression and to compare the mechanisms of constitutive release of the chymase, mMCP-1, and the chemokine, CCL2.Methods MMC homologues were generated by culturing bone marrow cells in the presence of TGF-β1, IL-3, IL-9 and stem cell factor (SCF). The intracellular distribution of mMCP-1 and CCL2 was examined by confocal microscopy. The involvement of the Golgi complex and of protein synthesis in the constitutive release of mMCP-1 and CCL2 was investigated using the Golgi-disrupting agent brefeldin A and cycloheximide to block protein synthesis. Secreted analytes were quantified by ELISA.Results mMCP-1 colocalized with Golgi matrix protein 130 but was most abundant in the granules, whereas CCL2 was not found in the granules but appeared to be located uniquely in the Golgi complex. Extracellular release of mMCP-1 was significantly inhibited (≈ 40%) by cycloheximide and by the Golgi-disrupting agent brefeldin A, indicating both continuous protein synthesis and transportation via the Golgi complex are required for optimal mMCP-1 secretion. A similar but more marked inhibitory effect with both compounds was demonstrated on the constitutive secretion of CCL2.Conclusion The culture conditions that promote mMCP-1 expression and release by MMC homologues also promote the expression and release of CCL2. Constitutive release involves de novo protein synthesis and requires a functional Golgi complex, suggesting that similar mechanisms of extracellular secretion operate for both mediators.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2222.2003.01571.x
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  • 3
    Electronic Resource
    Electronic Resource
    Transforming growth factor-bβ1 mediates coexpression of the integrin subunit aαE and the chymase mouse mast cell protease-1 during the early differentiation of bone marrow-derived mucosal mast cell homologues (2002)
    Oxford, UK : Blackwell Science Ltd
    Clinical & experimental allergy 32 (2002), S. 0 
    Details
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Mucosal mast cells (MMC) play a central role in gut hypersensitivities and inflammation. They are morphologically, biochemically and functionally distinct from their connective tissue counterparts. Massive hyperplasia of MMC occurs 7–10 days after intestinal infection with nematodes but it has never been possible to replicate this phenomenon in vitro.〈section xml:id="abs1-2"〉〈title type="main"〉Objective(1) To determine whether mouse bone marrow-derived mast cells (mBMMC) grown in the presence of transforming growth factor (TGF)-β1 could develop over the same time frame (7–10 days) as MMC in parasitized mice. (2) To compare the early expression of surface receptors (integrins αE and β7, c-kit and FcεR) with that of the MMC-specific granule chymase mouse mast cell protease-1 (mMCP-1).〈section xml:id="abs1-3"〉〈title type="main"〉MethodsMouse bone marrow cells were cultured in the presence of IL-9, IL-3 and Stem Cell Factor (SCF) with or without TGF-β1. mBMMC were quantified after toluidine blue or Leishmans' staining. Expression of MMC-specific mouse mast cell proteases was analysed by ELISA, immunohistochemistry and RT-PCR. Surface antigen expression was characterized by flow cytometry and confocal microscopy.〈section xml:id="abs1-4"〉〈title type="main"〉ResultsTGF-β1 promotes the development of abundant MMC-like mBMMC from bone marrow progenitor cells with kinetics, which closely parallel that seen in vivo. mRNA transcripts encoding mMCP-1 and -2 are readily detectable by day 4 ex vivo in cultures grown in the presence of TGF-β1. Between 30 and 40% and 75–90% of the cells in these cultures on days 4 and 7, respectively, have typical mast cell morphology, are c-kit+, FcεR+, integrin αEβ7+, and express and secrete abundant mMCP-1. The integrin αE subunit is coexpressed with mMCP-1.〈section xml:id="abs1-5"〉〈title type="main"〉ConclusionThe kinetics of mMCP-1+/αE+ mBMMC development, regulated by TGF-β1, are consistent with that seen in vivo in the parasitized intestine. The normally down-regulatory functions of TGF-β1 in haematopoiesis are superseded in this culture system by its ability to promote the early expression of αE and mMCP-1.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2222.2002.01233.x
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  • 4
    Electronic Resource
    Electronic Resource
    Local lung responses following local lung challenge with recombinant lungworm antigen in systemically sensitized sheep (2001)
    Oxford, UK : Blackwell Science, Ltd
    Clinical & experimental allergy 31 (2001), S. 0 
    Details
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background Chronic mast cell-mediated inflammation may contribute significantly towards the extensive tissue remodelling that is a feature of lungworm infection in ruminants. Understanding the factors that control tissue remodelling is a necessary step toward effective management and treatment of conditions that feature such pathology.Objective We sought to define in a novel ovine model system, the cellular, immune and mast cell phenotypic events that occur following local lung challenge with a recombinant protein antigen, DvA-1, derived from the ruminant lungworm nematode, Dictyocaulus viviparus.Methods Two spatially disparate lung segments in systemically sensitized sheep were challenged on three occasions with DvA-1 (3xDVA) and two further segments were challenged with saline (3xSAL). Two months after the third challenge, one of the two segments previously repeatedly challenged with DvA-1 was challenged again with DvA-1 (3xDVA:DVA) whilst the other was challenged with saline (3xDVA:SAL). A similar protocol was followed with the saline challenged segments (3xSAL:SAL and 3xSAL:DVA). Bronchoalveolar lavage fluid (BALF) (n = 16) and tissue (n = 3) were collected after the last challenge.Results Cellular changes 24 h after the fourth challenge were characterized by an increase in the absolute numbers of neutrophils and eosinophils in BALF from 3xDVA:DVA and 3xSAL:DVA segments. Local antibody production was implied through increased levels of antibody in both 3xDVA:DVA and 3xDVA:SAL segments, with the latter being unaffected by inflammation. Levels of active transforming growth factor beta-1 (TGF-β1) were significantly increased in 3xDVA:SAL segments and a trend towards an increase was apparent in 3xDVA:DVA segments. Total TGF-β1 levels were significantly correlated with eosinophil counts in all except the 3xDVA:SAL segments. Such changes in the bronchoalveolar space were complemented by increased ratios of sheep mast cell proteinase-1 expressing cells and tryptase expressing cells, to toluidine blue positive cells in airways from 3xDVA:DVA segments.Conclusion Mast cell phenotypic events occurring as a consequence of antigen challenge were limited to segments in which changes in BALF were characterized by neutrophil influx and increased local antibody production.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2222.2001.01225.x
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  • 5
    Electronic Resource
    Electronic Resource
    Mast cell specific proteases in rat brain: changes in rats with experimental allergic encephalomyelitis (1997)
    Springer
    Journal of neural transmission 104 (1997), S. 399-417 
    Details
    ISSN: 1435-1463
    Keywords: Rat mast cell protease I ; rat mast cell protease II ; experimental allergic encephalomyelitis ; histaminergic neurons ; histamine H3 receptor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Mast cell populations were identified within brain parenchyma by their specific proteases, using antibodies for immunohistochemistry and ELISAs, and riboprobes were developed for in situ hybridisation. Connective tissue mast cells expressing rat mast cell protease I (RMCPI) mRNA and immunoreactivity were observed in thalamus and showed no degranulation at 3, 8 and 13 days after induction of experimental allergic encephalomyelitis (EAE). Mucosal-like mast cells were clearly demonstrated in control rats by measuring RMCPII and by visualising cells expressing RMCPII mRNA and immunoreactivity. At day 13, but not 3 and 8 post immunisation, the number of RMCPII-expressing cells markedly increased in the EAE-induced group, mainly within brainstem and spinal cord close to inflammed blood vessels. The markers of histaminergic neurons were marginally affected 13 days after immunisation and the increase of [3H] histamine synthesis elicited by the H3-receptor antagonist, thioperamide, was not modified in any region of the brain. It is concluded that the cerebral RMCPII-expressing mast cells could play a role during EAE.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01277659
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  • 6
    Electronic Resource
    Electronic Resource
    Concomitant detection of mucosal mast cells and eosinophils in the intestines of normal and nippostrongylus-immune rats (1984)
    Springer
    Histochemistry and cell biology 81 (1984), S. 585-589 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The granules of mucosal mast cells (MMC) in the rat and man apparently poorly fixed with formaldehyde and special fixation techniques are normally used to demonstrate MMC glycosaminoglycans (GAG) in these two species. However such techniques do not permit the study of MMC granule enzyme cytochemistry or the demonstration of eosinophils. We have, therefore, examined some histochemical and immunocytochemical properties of MMC and eosinophils in normal and parasitised rats following various fixation procedures. Immersion fixation of rat intestine in 4% paraformaldehyde for 6 h not only facilitated the demonstration of MMC glycosaminoglycans with basic dyes but also permitted the concomitant staining of cosinophils with acidophilic dyes. A MMC granule-associated serine esterase was also demonstrated by enzyme cytochemistry and rat mast cell protease II was detected within MMC granules by immunocytochemistry. This new methodology obviates the requirement for separate fixation procedures in the identification and characterisation of MMC/eosinophil interactions in normal and parasitised rat intestinal mucosa.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00489539
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  • 7
    Electronic Resource
    Electronic Resource
    Mucosal mast cells are functionally active during spontaneous expulsion of intestinal nematode infections in rat (1984)
    [s.l.] : Nature Publishing Group
    Nature 312 (1984), S. 450-452 
    Details
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Following oral infection of PVG rats with 1,500 T. spiralis muscle larvae, most adult worms were expelled from the intestine between 9 and 12 days after infection (Fig. 1), elimination being complete by day 15. Mucosal mastocytosis commenced on day 6 and was maximal at day 12 (Fig. 1). No RMCP II ...
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1038/312450a0
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  • 8
    Electronic Resource
    Electronic Resource
    Characterization of whole-cell currents in mucosal and connective tissue rat mast cells using amphotericin-B-perforated patches and temperature control (1996)
    Springer
    Pflügers Archiv 432 (1996), S. 986-994 
    Details
    ISSN: 1432-2013
    Keywords: Key words Mast cells ; Whole-cell currents ; K+ channels ; Cl ; channels ; Inward rectifier
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  Rat mucosal type mast cells are thought to possess only a K+-selective inwardly rectifying (IRK) current in the resting state. We used rat-bone-marrow-derived mast cells (BMMCs) as a model of mucosal mast cells and recorded whole-cell membrane currents from cells perforated with amphotericin B. Under these conditions, both inwardly rectifying (IR) and outwardly rectifying (OR) currents were observed. The reversal potential and conductance of the IR current depended on the extracellular K+ concentration, indicating that the channel was K+ selective. The OR current was not affected by changes in extracellular K+ concentration, but lowering extracellular Cl–concentration reduced the conductance and shifted the reversal potential in a positive direction. The OR current was not affected by K+ channel blockers, but was reversibly blocked by the chloride channel blocker 4,4’-diisothiocyanato-2,2’-stilbenedisulphonate (DIDS), again indicating a Cl–conductance. The IRK current was also detected in the majority of cells using the conventional whole-cell recording configuration at room temperature. In contrast, the ORCl current was only observed in 7% of recordings made at room temperature with the conventional whole-cell voltage-clamp mode, but was detected in 66% of cells if the bath temperature was increased and the integrity of the cell’s cytoplasm was preserved by using the perforated-patch technique. Under similar conditions, the ORCl current was also present in rat peritoneal mast cells, a connective tissue phenotype previously thought to have no whole-cell currents in the resting state. The role of this current and factors affecting its activation are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004240050226
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  • 9
    Electronic Resource
    Electronic Resource
    Fixation and tissue preservation for antibody studies (1972)
    Springer
    Journal of molecular histology 4 (1972), S. 305-320 
    Details
    ISSN: 1573-6865
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Synopsis The methods of fixation and preparation of lymphoid tissues for the immuno-enzyme technique are reviewed. For this technique an enzyme is used first as an antigen and then as a marker to demonstrate its specific antibody. A variety of commonly employed fixatives satisfactorily conserve tissues for the light microscopic detection of antibody but, for electron microscopy, glutaraldehyde or formaldehyde or both are the fixatives of choice. The main technical problem for electron microscopy is to reduce the size of the tissue fragments sufficiently so that the enzymes and their substrates permeate through the fixed tissues. The merits and short-comings of the different preparative techniques are examined and it is shown that the most reproducible results are obtained with 40 μm frozen sections. Some of the problems of non-specific staining arising from fixation procedures, as well as endogenous enzyme activity, are discussed. The evidence for and against antibody inactivation by fixation and enzyme inactivation by interaction with its specific antibody is reviewed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01005006
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