scholarly journals Ultrastructural localization of basic proteins in cytoplasmic granules of rat eosinophils and mast cells.

1980 ◽  
Vol 28 (3) ◽  
pp. 238-244 ◽  
Author(s):  
P F Pimenta ◽  
M A Loures ◽  
W de Souza
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Basic Protein ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Cytoplasmic Granules ◽  
Basic Proteins ◽  
Intense Reaction ◽  
The Matrix ◽  
Ammoniacal Silver

The postformalin ammoniacal silver (AS) and the ethanolic phosphotungstic acid (EPTA) methods were used to localize basic protein at the ultrastructural level in cytoplasmic granules of rat eosinophils and mast cells isolated using a Metrizamide gradient. Intense reaction was seen in the granules of EPTA-treated eosinophils. Following incubation of the cells for 2 hr in EPTA alone, the matrix was stained. After longer incubation (10 hr), however, both the matrix and core were stained. Cytoplasmic granules of the mast cell show a slight or negative reaction with EPTA. With the AS technique, a large number of silver particles were seen in the nucleus of both eosinophils and mast cells. The mast cell cytoplasmic granules showed intense reaction, while those from eosinophils showed no clear reaction. Acetylation of the cells under conditions sufficient to block most free amino groups prio to EPTA or AS treatment greatly reduced (EPTA) or abolished (AS) the reaction. The results indicate 1) that eosinophil granules contain basic proteins both in the matrix and the core, 2) that the mast cell granules contain a basic protein (probably the alpha-chymotrypsin-like enzyme), which reacts strongly with AS, and 3) that the AS and EPTA methods have different specificities.

scholarly journals Ultrastructural localization of basic proteins in Trypanosoma cruzi.

1978 ◽  
Vol 26 (5) ◽  
pp. 349-358 ◽  
Author(s):  
T Souto-Padrón ◽  
W De Souza
Keyword(s):  
Trypanosoma Cruzi ◽  
Phosphotungstic Acid ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Basic Proteins ◽  
Intense Reaction ◽  
Cytoplasmic Vacuoles ◽  
Ammoniacal Silver

The postformalin ammoniacal silver (AS) and the ethanolic phosphotungstic acid (EPTA) techniques were applied in epimastigote and trypomastigote forms of the pathogenic protozoa Trypanosoma cruzi to detect basic proteins at the ultrastructural level. With both techniques, reaction was observed in the nucleus and in some cytoplasmic vacuoles. In the kinetoplast of epimastigotes, reaction was observed only at its periphery. In trypomastigotes, however, an intense reaction was observed in the spherical kinetoplast. With the ethanolic phosphotungstic acid technique, reaction was also observed in ribosomes and at the peripheral doublet microtubules of the flagellum. The filaments which form the paraflagellar structure did not react.

Ultrastructural localization of cationic proteins in human polymorphonuclear leukocytes

1978 ◽  
Vol 30 (1) ◽  
pp. 21-35
Author(s):  
W.J. Brown ◽  
E.M. Wood
Keyword(s):  
Polymorphonuclear Leukocytes ◽  
Osmium Tetroxide ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Cationic Proteins ◽  
Cytoplasmic Granules ◽  
Mediators Of Inflammation ◽  
Human Polymorphonuclear Leukocytes ◽  
Ammoniacal Silver ◽  
Neutral Proteases

The present investigation is concerned with the use of the post-formalin ammoniacal silver reaction to detect the arginine-rich cationic proteins in human polymorphonuclear leukocytes at the ultrastructural level. These proteins appear to function as neutral proteases in antibacterial action and as mediators of inflammation. Originally, the ammoniacal silver reaction relied upon primary fixation in dilute formalin which prevented optimum fixation of tissues. This study shows that by using the proper sequence of glutaraldehyde fixation and the ammoniacal silver solution in conjunction with osmium tetroxide treatment, better fixation of the tissue and localization of the ammoniacal silver reaction can be achieved. Also, the ammoniacal silver reaction in human polymorphonuclear leukocytes is exclusively located in the large, crystalline cytoplasmic granules of eosiniphils. All other cytoplasmic granules of neutrophils, eosinophils, and basophils were found to be devoid of the ammoniacal silver reaction product. These results are contrary to previously published experimental data. Possible explanations for this discrepancy are discussed.

Ultrastructural localization of cationic proteins in cytoplasmic granules of chicken and rabbit polymorphonuclear leukocytes

1975 ◽  
Vol 17 (1) ◽  
pp. 79-94
Author(s):  
E.K. Macrae ◽  
J.K. Spitznagel
Keyword(s):  
Polymorphonuclear Leukocytes ◽  
Intracellular Localization ◽  
Ultrastructural Localization ◽  
Cationic Protein ◽  
Cationic Proteins ◽  
Cytoplasmic Granules ◽  
Electron Dense Deposit ◽  
Dense Deposit ◽  
Neutrophilic Polymorphonuclear Leukocytes ◽  
Ammoniacal Silver

Cytoplasmic granules known to contain cationic arginine-rich proteins can be identified by the ammoniacal silver reaction (ASR) which provides a cytochemical marker detectable under the electron microscope. Only the large rod-shaped granules of the chicken polymorphonuclear leukocytes (heterophils) and the large spherical azurophilic granules of the rabbit neutrophilic polymorphonuclear leukocytes show the ASR product as a discrete particulate electron-dense deposit. The other smaller granules are devoid of reaction product, as are membranes and mitochondria. The intracellular localization of the ASR product, as are membranes and mitochondria. The intracellular localization of the ASR product on the large granules coincides with the ASR product localization on the same isolated granule populations, when the ammoniacal silver reaction is applied to these granules after their separation by sucrose-density gradients. The cationic proteins may have intraleukocytic bacteriolytic properties, since ASR product, presumably indicating cationic protein from discharged granules, appears to surround ingested bacteria within cytoplasmic phagosomes.

scholarly journals Peroxidase activity in human cutaneous mast cells: an ultrastructural demonstration.

1984 ◽  
Vol 32 (6) ◽  
pp. 573-578 ◽  
Author(s):  
L M Escribano ◽  
L C Gabriel ◽  
T Sainz ◽  
A Rocamora ◽  
J M Arrazola ◽  
...  
Keyword(s):  
Mast Cell ◽  
Enzyme Activity ◽  
Mast Cells ◽  
Peroxidase Activity ◽  
Ultrastructural Level ◽  
Prostaglandin Synthesis ◽  
Cell Disease ◽  
Mast Cell Disease ◽  
Cytochemical Technique ◽  
The Relationship

An intense and reproducible peroxidase staining in the cutaneous mast cells of two patients with systemic mast cell disease and urticaria pigmentosa is demonstrated at the ultrastructural level. This enzyme activity was demonstrated by use of a cytochemical technique employing 3,3'- diaminobenzicine (DAB) as an oxidizable substrate, after fixation by a tannic acid-aldehyde mixture. Enzyme activity was localized in the perinuclear cisterna and strands of endoplasmic reticulum. Granules appeared unreactive. This peroxidase activity appears sensitive to fixation by aldehydes; it is inhibited by 3-amino-1,2,4-triazole (AMT) and by lack of H2O2 or DAB in the incubation medium. These characteristics are fundamentally different from the peroxidase activity of basophils, and the demonstration of this enzyme is therefore not a further argument for a common ontogenetic origin of both cells. On the other hand, the cytochemical characteristics of this enzyme are very similar to those of platelet peroxidase (P-PO), which has been connected to the synthesis by platelets of prostaglandins. Since the mast cell is known to generate prostaglandins, the relationship between the enzyme described and prostaglandin synthesis by mast cells is discussed.

scholarly journals Immunomagnetic Isolation of Rat Bone Marrow-derived and Peritoneal Mast Cells

1997 ◽  
Vol 45 (12) ◽  
pp. 1715-1722 ◽  
Author(s):  
Maria Celia Jamur ◽  
Ana Cristina G. Grodzki ◽  
Andrea N. Moreno ◽  
William D. Swaim ◽  
Reuben P. Siraganian ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Mast Cell ◽  
Mast Cells ◽  
Cell Types ◽  
Ultrastructural Level ◽  
Immunomagnetic Separation ◽  
Specific Cell ◽  
Morphological Examination ◽  
Immunomagnetic Isolation ◽  
Rat Bone

Mast cells are difficult to purify from heterogeneous cell populations and to preserve, especially for pre-embedding immunostaining at the ultrastructural level. We have developed a technique that permits the isolation of a pure population of mast cells suitable for immunocytochemical studies. A rat mast cell-specific monoclonal antibody (MAb AA4) conjugated to tosylactivated Dynabeads 450 was used to immunomagnetically separate mast cells from rat bone marrow and peritoneal cell suspensions. Approximately 85% of the mast cells were recovered in the positive population that comprised virtually pure mast cells. After microwave fixation, morphological examination showed that the cells were intact and retained their ultrastructural detail. Mast cells in all stages of maturation were immunolabeled with a panel of antibodies after immunomagnetic separation. The combination of immunomagnetic separation followed by immunostaining should prove useful for the study of mast cell maturation and for the characterization of other specific cell types that are present in tissues in only limited numbers.

scholarly journals Polyethyleneimine as a contrast agent for ultrastructural localization and characterization of proteoglycans in the matrix of cartilage and bone.

1991 ◽  
Vol 39 (3) ◽  
pp. 331-340 ◽  
Author(s):  
Y M Sauren ◽  
R H Mieremet ◽  
C G Groot ◽  
H K Koerten ◽  
J P Scherft
Keyword(s):  
Bone Matrix ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Cartilage Matrix ◽  
Proteoglycan Synthesis ◽  
Positive Material ◽  
Calcified Cartilage ◽  
The Matrix ◽  
Proteoglycan Aggregates

We examined the presence of proteoglycans in the extracellular matrix of cartilage and bone in fetal mouse radii at the ultrastructural level, using the cationic dye polyethyleneimine (PEI). After staining with this dye, the proteoglycans appeared as granules in the uncalcified bone matrix and as extended winding structures in the cartilage matrix. PEI-positive material was removed after treatment of the tissue with chondroitinase ABC. Inhibition of the proteoglycan synthesis by beta-D-xyloside resulted in smaller PEI-positive windings in the cartilage matrix. These observations suggest that the winding, PEI-positive structures represent proteoglycan aggregates. No loss of PEI-positive material in the calcified cartilage matrix was seen, suggesting that proteoglycans do not need to be removed to make the matrix calcifiable.

Ultrastructural evidence for bladder MAST cell degranulation and their imminent association with nerve fibers in patients with interstitial cystitis

1994 ◽  
Vol 52 ◽  
pp. 264-265
Author(s):  
F. He ◽  
M. Hofmeister ◽  
T. Ratliff ◽  
M. Becich
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Interstitial Cystitis ◽  
Ultrastructural Level ◽  
Nerve Fibers ◽  
Morphological Description ◽  
Electron Microscopic ◽  
Human Bladder ◽  
Surgical Biopsy ◽  
Transmission Electron

The ultrastructure of human mast cells (HMCs) in various diseases has been well documented; however, detailed morphological description of human bladder mast cells (HBMCs) in patients with interstitial cystitis (IC) is incomplete. The present study was undertaken to reveal any morphological modifications of human bladder mast cells (HBMCs) from the IC patients and to investigate the spatial relationships between nerve fibers and these mast cells at the ultrastructural level.Fresh-fixed surgical biopsy specimens from IC patients as well as paraffin blocks of IC retrieved through our medical record archival system were processed with routine and deparaffinization methods for transmission electron microscopy (1). No differences in mast cell morphology were noticed between the fresh-fixed or paraffin embedded tissues.The general electron microscopic appearances of the HBMCs in IC located in both mucosa and muscle layers were similar to those of HMCs observed in other sites where mastocytosis may occur due to various pathological processes (Fig. 1).

Stem Cell Factor Influences Mast Cell Mediator Release in Response to Eosinophil-Derived Granule Major Basic Protein

Blood ◽  
1998 ◽  
Vol 92 (3) ◽  
pp. 1055-1061
Author(s):  
Glenn T. Furuta ◽  
Steven J. Ackerman ◽  
Lei Lu ◽  
Rachel E. Williams ◽  
Barry K. Wershil
Keyword(s):  
Mast Cell ◽  
Stem Cell ◽  
Mast Cells ◽  
Stem Cell Factor ◽  
Basic Protein ◽  
Mouse Bone Marrow ◽  
Major Basic Protein ◽  
Mast Cell Line ◽  
American Society ◽  
Factor Α

Stem cell factor (SCF) is an important mast cell growth, differentiation, and survival factor. We investigated whether SCF influenced the response of mouse mast cells to an IgE-independent stimulus, eosinophil-derived granule major basic protein (MBP). Mouse bone marrow cultured mast cells (BMCMC) were derived in either concanavalin-stimulated mouse spleen conditioned medium (CM) or SCF. The cloned growth, factor-independent mast cell line Cl.MC/C57.1 was also studied. BMCMC in SCF exhibited cytochemical staining properties, protease and histamine content, and increased serotonin uptake consistent with more mature differentiated mast cells as compared with BMCMC in CM or Cl.MC/ C57.1 cells. BMCMC in SCF released serotonin,14C-labeled arachidonic acid metabolites and tumor necrosis factor-α (TNF-α) on stimulation with MBP, while no response was seen from either BMCMC in CM or Cl.MC/C57.1 cells. All three mast cell populations released mediators on stimulation with the cationic MBP analog, poly-L-arginine, indicating that the cationic charge did not explain the selective response of BMCMC in SCF to eosinophil-derived granule MBP. These findings show that SCF significantly influences mast cell differentiation and the responsiveness of mast cells to eosinophil-derived granule MBP. © 1998 by The American Society of Hematology.

scholarly journals Mast Cell Chymase and Kidney Disease

2020 ◽  
Vol 22 (1) ◽  
pp. 302
Author(s):  
Shamila Vibhushan ◽  
Manuela Bratti ◽  
Juan Eduardo Montero-Hernández ◽  
Alaa El Ghoneimi ◽  
Marc Benhamou ◽  
...  
Keyword(s):  
Mast Cell ◽  
Angiotensin Ii ◽  
Mast Cells ◽  
Kidney Disease ◽  
Inflammatory Response ◽  
Inflammatory Mediator ◽  
Total Proteins ◽  
Cytoplasmic Granules ◽  
Physiological Processes ◽  
Mast Cell Chymase

A sizable part (~2%) of the human genome encodes for proteases. They are involved in many physiological processes, such as development, reproduction and inflammation, but also play a role in pathology. Mast cells (MC) contain a variety of MC specific proteases, the expression of which may differ between various MC subtypes. Amongst these proteases, chymase represents up to 25% of the total proteins in the MC and is released from cytoplasmic granules upon activation. Once secreted, it cleaves the targets in the local tissue environment, but may also act in lymph nodes infiltrated by MC, or systemically, when reaching the circulation during an inflammatory response. MC have been recognized as important components in the development of kidney disease. Based on this observation, MC chymase has gained interest following the discovery that it contributes to the angiotensin-converting enzyme’s independent generation of angiotensin II, an important inflammatory mediator in the development of kidney disease. Hence, progress regarding its role has been made based on studies using inhibitors but also on mice deficient in MC protease 4 (mMCP-4), the functional murine counterpart of human chymase. In this review, we discuss the role and actions of chymase in kidney disease. While initially believed to contribute to pathogenesis, the accumulated data favor a more subtle view, indicating that chymase may also have beneficial actions.

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