Mast cell heterogeneity between two different species of Hoplias sp. (Characiformes: Erythrinidae): response to fixatives, anatomical distribution, histochemical contents and ultrastructural features

Mast cells from two erythrinid species: Hoplias malabaricus and Hoplias lacerdae, were studied in several tissues throughout the body using light and electron microscopy. Mast cells were found in all organs studied, but were especially abundant in the gastrointestinal tract, and were always in association with connective tissue. These cells showed different characteristics between the two species studied, like varied morphology, anatomical distribution, density, basophilic/eosinophilic staining and heparin content. In H. malabaricus, the tissues fixed with Helly's solution contained mast cells that were basophilic, metachromatic and had heparin in their cytoplasmic granules, while the tissues fixed with Karnovsky's solution contained eosinophilic and orthochromatic mast cells in which heparin was not detected. In H. lacerdae, the use of both fixatives resulted in mast cells that were eosinophilic, orthochromatic, with no identifiable heparin content. Exclusively in H. malabaricus oesophagus, the mast cells were additionally seen among the epithelial cells. The ultrastructural studies performed in hindgut fixed with Karnovsky's solution revealed that the cytoplasmic granules seen in H. lacerdae mast cells were better preserved than in H. malabaricus mast cells. The latter had electron-lucent granules that were often merged, forming channels. The present study demonstrated that mast cells from two species belonging to the same genus or even mast cells from the same species but under different fixatives can present heterogeneous characteristics, possibly due to their functional properties or to their sensitivity to fixatives.     

牛、羊胆囊内肠道菌的分离鉴定

本文对育肥屠宰的牛、羊胆囊进行了细菌学调查,其胆囊内细菌的污染情况是:被检的79例牛羊胆囊,有31例带菌,带菌率为31/79。在32例牛胆囊,有12例带菌,带菌率为12/32;47例羊胆囊,有19例带菌,带菌率为19/47。共分离出菌37株(牛2例、羊4例同时检出2株菌,其余均为1株)。按照Bergey氏《细菌鉴定手册》第8版的分类系统,经鉴定这些菌分别属5科8属中的10种菌(内含1株酵母菌及1株未定菌):大肠埃希氏菌、肠球菌、产气肠杆菌、普通变形杆菌、柠檬酸细菌,粪产硷杆菌、乳酸杆菌、表皮葡萄球菌。调查结果表明:正常情况下牛、羊胆囊内是无菌的。胆囊内检出的细菌是属污染菌,且均为肠道菌。     

Immunocytochemical Localization of Aromatase in The Ovary of Superovulated Cattle,Pigs and Sheep

The localization of aromatase, an enzyme converting androgens to estrogen, in the ovaries of superovulated cattle, pigs and sheep was studied immunocytochemically in the preovulatory and postovulatory period using anti-human placental aromatase cytochrome P-450 antiserum. Immunostaining for aromatase was detected in the granulosa cells of preovulatory follicles of all species studied. Theca interna cells were stained in preovulatory follicles in the pig but not in cattle and sheep. Interstitial gland cells, cumulus cells and oocytes were unstained in all species. In cattle and pig the corpora lutea were unstained whereas they displayed staining in the sheep. Preantral and small antral follicles were unstained during both the preovulatory and postovulatory period in all species. It is concluded that granulosa cells of preovulatory follicles are the main residence for aromatase activity in superovulated cattle, pig and sheep, whereas the activity of theca interna and corpora lutea is species specific.     

Histochemical and morphological characterization of migrating mast cells in the bovine gallbladder epithelium

Summary This paper reports the existence of granulated cells in the bovine gallbladder epithelium. The histochemical studies performed on these cells showed that their cytoplasmic granules contain heparin. Optical- and electron microscopic observations demonstrated that these mast cells migrated from the adjacent connective tissue, and crossed the basal membrane in order to enter the epithelium and reach the luminal surface.     

Mast cell glycosaminoglycans

Mast cells contain granules packed with a mixture of proteins that are released on degranulation. The proteoglycan serglycin carries an array of glycosaminoglycan (GAG) side chains, sometimes heparin, sometimes chondroitin or dermatan sulphate. Tight packing of granule proteins is dependent on the presence of serglycin carrying these GAGs. The GAGs of mast cells were most intensively studied in the 1970s and 1980s, and though something is known about the fine structure of chondroitin sulphate and dermatan sulphate in mast cells, little is understood about the composition of the heparin/heparan sulphate chains. Recent emphasis on the analysis of mast cell heparin from different species and tissues, arising from the use of this GAG in medicine, lead to the question of whether variations within heparin structures between mast cell populations are as significant as variations in the mix of chondroitins and heparins.     

A comparative study of lymph node mast cell populations in five marsupial species.

In order to determine whether different subpopulations of mast cells exist, mast cells of mandibular and axillary lymph nodes from five species (Didelphis aurita, Metachirus nudicaudatus, Philander opossum, Marmosops incanus and Gracilinanus agilis) of South American marsupials were studied. Our results showed that mast cells present in the connective tissue of the capsule and septa (CTMC) were similar to those described for eutherian mammals. However, a population of mast cells that was present in the lymphatic sinuses and bathed by the lymph, plus in direct contact with granulocytes, lymphocytes, macrophages, and reticular cells, were morphologically and histochemically different from the CTMC. In the five species studied, these cellular types, called lymphatic-sinus mast cells (LSMC), had a lower concentration of intragranular heparin and, in four of the five species, the cytoplasmic granules appeared to be larger than those in CTMC. Although LSMC have been rarely described in eutherian mammals, it was verified, in this study, that LSMC are nevertheless present in lymphatic sinuses of the five metatherian species studied. These observations suggest that the presence of LSMC may be a characteristic of the marsupials and important in the immune response and adaptive success of the Didelphidae.     

Immunologic mast cell-mediated responses and histamine release are attenuated by heparin.

Heparin has been shown to act as a competitive inhibitor of inositol 1,4,5-triphosphate (InsP3) receptors in various cell types. Because InsP3 is one of the second messengers involved in stimulus-secretion coupling in mast cells, it is possible that heparin may inhibit mast cell-mediated reactions. Therefore, in allergic sheep, we tested this hypothesis in two mast cell-mediated reactions induced by immunologic and nonimmunologic stimuli: immediate cutaneous reaction (ICR) and acute bronchoconstrictor response (ABR). In 12 sheep allergic to Ascaris suum antigen, the surface area of the skin wheal was determined 20 min after intradermal injection (0.05 ml) of increasing concentrations of specific antigen, compound 48/80, and histamine, without and after pretreatment with heparin (100, 300, or 1,000 U/kg i.v.). Antigen, compound 48/80, and histamine produced concentration-dependent increases in ICR. Heparin "partially" inhibited the ICR to antigen and compound 48/80 in a dose-dependent manner without modifying the ICR to histamine. The heparin preservative benzyl alcohol was ineffective. In 11 additional sheep, specific lung resistance was measured before and after inhalation challenges with antigen, compound 48/80, and histamine without and with aerosol heparin pretreatment (1,000 U/kg). Heparin blocked the antigen- and compound 48/80-induced bronchoconstriction without modifying the airway effects of histamine. In isolated human uterine mast cells, heparin inhibited the anti-immunoglobulin E- but not the calcium ionophore- (A23187) induced histamine release. These data suggest that heparin inhibits the ICR and ABR induced by stimuli that produce immunologic and nonimmunologic mast cell degranulation without attenuating the effects of histamine.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in numbers and heparin content of peritoneal fluid mast cells of growing rats measured by flow cytofluorometry

A cytofluorometric method, based on berberine staining of mast cell heparin, was used for flow cytofluorometric counting and heparin quantitation of mast cells in crude peritoneal suspensions of growing rats. The automatic flow cytofluorometric counting of mast cells correlated well with hemocytometer cell counts. The mean mast cell heparin content obtained by flow cytofluorometry showed good agreement with such obtained by cytofluorometry of microscopically identified mast cells. The number of peritoneal mast cells and the mean mast cell heparin content was found to increase as the animals grew older. The results of the microscope fluorometric measurements suggested that the heparin content was normally distributed within mast cell populations of both young and old rats. However, the heparin distributions obtained by flow cytofluorometry were often positively skewed but did not fulfill the condition of the log-normal distribution.     

Protein content, dry mass and chemical composition of individual mast cells related to body growth.

Recently developed quantitative microscopical techniques were used to study relations between body growth and protein content as well as dry mass of individual mast cells. Since previous studies had shown an age-related increase of mast cell content of 5-hydroxytryptamine (5-HT) and heparin, these mast cell components were also included in the present study. The cells were obtained from the peritoneal cavity of rats aged 44--269 days (body weights 189--610 g). All studied mast cell parameters showed an increase that was related to the growth of the animals. The dry mass increased 60%, protein 50%, heparin 50% but 5-HT increased as much as 260% during the studied growth period. There was a mutual and linear correlation between all studied mast cell parameters. Population studies, based on large scale measurements of individual mast cells from young and adult rats, were made. These studies showed that histograms of 5-HT content, protein content and dry mass of individual mast cells were skewed with a tail towards higher values and approximately lognormal. On the other hand, the frequency distribution of heparin content of individual mast cells was approximately normal.     

Mast cells are present in epithelial layers of different tissues of the mollusc Anomalocardia brasiliana. In situ characterization of heparin and a correlation of heparin and histamine concentration

Heparin, other sulphated glycosaminoglycans and histamine were extracted from various dissected organs of Anomalocardia brasiliana, a mollusc from the South Atlantic, and quantified. A good correlation between heparin and histamine content was found in the labial palp, intestine, ctenidium, mantle and foot tissues. The tissue location of metachromatic cells, putatively containing heparin, was identified histologically with Alcian Blue, Toluidine Blue, Masson trichrome, Haematoxylin-Eosin and PAS. Except for the foot, cells containing metachromatic granules were found in the epithelium surfaces of all the organs analysed. An in situ identification of heparin using nitrous acid and heparinase degradation has established unequivocally the presence of this compound in the metachromatic cells. The location of 'mast-like' cells at the epithelium surface of mollusc tissues exposed to the environment are very similar to the distribution of mammalian and other vertebrate mast cells and gives support to the suggestion for a role of mast cells in defense mechanisms.     

Protein content,dry mass and chemical composition of individual mast cells related to body growth

Summary Recently developed quantitative microscopical techniques were used to study relations between body growth and protein content as well as dry mass of individual mast cells. Since previous studies had shown an age-related increase of mast cell content of 5-hydroxytryptamine (5-HT) and heparin, these mast cell components were also included in the present study. The cells were obtained from the peritoneal cavity of rats aged 44–269 days (body weights 189–610 g). All studied mast cell parameters showed an increase that was related to the growth of the animals. The dry mass increased 60%, protein 50%, heparin 50% but 5-HT increased as much as 260% during the studied growth period. There was a mutual and linear correlation between all studied mast cell parameters. Population studies, based on large scale measurements of individual mast cells from young and adult rats, were made. These studies showed that histograms of 5-HT content, protein content and dry mass of individual mast cells were skewed with a tail towards higher values and approximately lognormal. On the other hand, the frequency distribution of heparin content of individual mast cells was approximately normal.Supported by grants from the Swedish Medical Research Council, Project no 2235     

Heparin modulates the growth and adherence and augments the growth-inhibitory action of TNF-alpha on cultured human keratinocytes

Previous works suggest the involvement of mast cells in the epithelialization of chronic wounds. Since heparin is a major mediator stored in the secretory granules of mast cells, the purpose of this work was to elucidate the function of heparin in epithelialization using in vitro culture models. For this, low- and high-calcium media in monolayer and epithelium cultures of keratinocytes were used. Also, an assay based on keratinocyte adherence onto plastic surface was used as well. Heparin (0.02-200 microg/ml) inhibited keratinocyte growth in a non-cytotoxic and dose-dependent manner in low- and high-calcium media, Keratinocyte-SFM and DMEM, in the absence of growth factors and serum. Also, heparin inhibited the growth of keratinocyte epithelium in the presence of 10% fetal calf serum and DMEM. Instead, in the presence of Keratinocyte-SFM and growth factors, heparin at 2 microg/ml inhibited the growth by 18% but at higher heparin concentrations the inhibition was reversed to baseline. TNF-alpha is another preformed mediator in mast cell granules and it inhibited keratinocyte growth in monolayer and epithelium cultures. Interestingly, heparin at 2-20 microg/ml augmented or even potentiated this growth-inhibitory effect of TNF-alpha. The association of TNF-alpha with heparin was shown by demonstrating that TNF-alpha bound tightly to heparin-Sepharose chromatographic material. However, heparin could not augment TNF-alpha-induced cell cycle arrest at G0/G1 phase or intercellular adhesion molecule-1 expression in keratinocytes. In the cell adherence assay, heparin at 2 microg/ml inhibited significantly by 12-13% or 33% the adherence of keratinocytes onto the plastic surface coated with fibronectin or collagen, respectively, but this inhibition was reversed back to baseline at 20 or 200 microg/ml heparin. Also, heparin affected the cell membrane rather than the protein coat on the plastic surface. In conclusion, heparin not only inhibits or modulates keratinocyte growth and adherence but it also binds and potentiates the growth-inhibitory function of TNF-alpha.     

Histochemical studies on mast cells in cattle skin

Synopsis Cells in the skin of cattle which gave a green fluorescence after formaldehyde treatment could be stained orange with Acridine Orange and blue with Astrablau. It is concluded that these cells are mast cells containing heparin and a catecholamine, probably dopamine.     

The role of mast cells in vascular permeability disorders in rats subjected to immobilization stress]

Disturbances of vascular permeability were studied by the "vascular labeling" technique in the mesentry during the 24-hour immobilization of rats. Administration of dimebolin (an antihistaminic preparation) decreased the number of labeled vessels and labeling intensity. This effect was expressed in the presence of mast cells only and was accompanied by the mast cell degranulation. The authors suppose that the mast cells contain a substance preventing the disturbance of vascular permeability and released during degranulation. Such substance might be heparin. Experiments showed that small doses of heparin failed to produce such effect. These results allowed one to conclude that mast cells played a double role in the mechanisms of disturbance of vascular permeability during immobilization--the damaging (by the action of histamine and serotonine) and the protective (by the released heparin) action.     

Composition of the rumen ciliate population in experimental herds of cattle and sheep in Lethbridge, Alberta, Western Canada

Rumen ciliate populations were surveyed in 11 Holstein cattle and 6 sheep in Lethbridge (Alta., Canada) to determine species distribution in this Western Canadian environment. A total of 28 ciliate species were identified in cattle and 17 in sheep. The average total number of ciliates per millilitre of rumen content was 6.9 X 10(4) in cattle and 1.9 X 10(5) in sheep. The average number of species per host was 20.5 in cattle and 13.8 in sheep. Of the ciliate species detected, species of Entodinium appeared most frequently both in cattle and in sheep. Diplodinium polygonale, Eodinium lobatum, Eo. monolobum, Eremoplastron rostratum, Ostracodinium clipeolum, Os. mammosum, and Ophryoscolex purkynjei were not detected in sheep. In contrast, Ophryoscolex caudatus was not found in cattle. These data indicate that the ciliate faunas of cattle and sheep in this Western Canadian environment are similar to those found in Japan.     

Reactions of cells of nasal and sinusoidal mucosa of goats and sheep naturally infected by Oestrus ovis Linné 1758 (Diptera: Oestridae)]

OEstrosis is a very common myiasis of sheep and goats in mediterranean and tropical countries. Goats are suitable hosts for OEstrus ovis but the parasitic burden remain lower than in sheep. Cellular responses (mucous and serous mast cells, eosinophis and globules leucocytes) were measured in 30 infected and 30 non infected sheep and in 23 infected and 24 non infected goats. The presence of OEstrus ovis larvae led to an important inflammatory response in sheep and in goats as well. Furthermore, the intensity of the cellular response correlated with the larva burden, specially with globules leucocytes and eosinophils. Nevertheless, huge differences occurred between sheep and goats responses even in similar larval burden range. Infected sheep showed larger counts of mucous mast cells than goats, the differences were smaller in serous mast cells and eosinophils and no difference was detected in globules leucocytes (intraepithelial mast cells) counts between the two hosts species. These results were compared with those obtained in gastro-intestinal strongyles infections.     

Electrolyte absorption by gallbladders: models of transport.

A model of electrolyte absorption by gallbladder epithelium has been presented previously on the basis of studies on gallbladders of 12 species, including fishes, frogs, toad, turtle, guinea pig, rabbit, cat and dog. This model incorporates several physiologic and morphologic characteristics common to other transporting epithelia (e.g., intestine) such as energy-dependent solute pumps, osmotically-induced water flow into the lateral intercellular space and bulk flow of fluid driven by hydrostatic pressure along the lateral space toward the basement membrane. Because the transepithelial PD across the gallbladders of each of these species was near zero under most experimental conditions, the active transport mechanism (or, “pump”) at the basolateral membrane was considered to move Na and Cl in a coupled, one-for-one manner. The carrier mechanism was postulated to have a binding site for Na and one for Cl; it would function only if both sites were filled.Gallbladders from six other species investigated more recently (including man, monkeys, goose and Necturus) have serosa-positive transepithelial PDs of 2–8 mV. The possibility was suggested that rheogenic Na transport from mucosa to serosa might account for the PD in this group of tissues and the original model of transport would be inappropriate. This review will explore the possibility that a single model of electrolyte transport accounts for the data collected on gallbladders with PDs near zero and those having significant transepithelial PDs.An important finding which helps to reconcile the experimental observations on the two groups of gallbladders was the demonstration of coupled flux of Na and Cl from the mucosal solution into the epithelial cells. It appears that this rigid coupling of Na and Cl influx accounts for the lack of a significant PD in gallbladders of those species investigated in the earlier studies, and that rheogenic Na transport may be a property common to gallbladders of all species.     

Mast cells are present in epithelial layers of different tissues of the mollusc <Emphasis Type="Italic">Anomalocardia brasiliana</Emphasis>. <Emphasis Type="Italic">In situ</Emphasis> characterization of heparin and a correlation of heparin and histamine concentration

Heparin, other sulphated glycosaminoglycans and histamine were extracted from various dissected organs of Anomalocardia brasiliana, a mollusc from the South Atlantic, and quantified. A good correlation between heparin and histamine content was found in the labial palp, intestine, ctenidium, mantle and foot tissues. The tissue location of metachromatic cells, putatively containing heparin, was identified histologically with Alcian Blue, Toluidine Blue, Masson trichrome, Haematoxylin–Eosin and PAS. Except for the foot, cells containing metachromatic granules were found in the epithelium surfaces of all the organs analysed. An in situ identification of heparin using nitrous acid and heparinase degradation has established unequivocally the presence of this compound in the metachromatic cells. The location of 'mast-like' cells at the epithelium surface of mollusc tissues exposed to the environment are very similar to the distribution of mammalian and other vertebrate mast cells and gives support to the suggestion for a role of mast cells in defense mechanisms.     

Comparative studies on the distribution of rhodanese in different tissues of domestic animals.

1. The activity of rhodanese in different tissues of some domestic animals was measured. 2. Rhodanese was present in all tissues studied. 3. The activity of rhodanese in most tissues of sheep was higher than other animals studied. 4. In sheep and cattle the epithelium of rumen, omasum and reticulum were the richest sources of rhodanese. Significant activity of rhodanese was also present in liver and kidney. 5. In camel the liver contained the highest level of rhodanese followed by lung and rumen epithelium. Camel liver contained a third of the activity of sheep liver. 6. Equine liver had a third of the activity of sheep liver. Other tissues showed low levels of rhodanese activity. 7. Dog liver contained only 4% of the activity of sheep liver. In this animal, brain was the richest source of rhodanese. 8. The results are discussed in terms of efficacy of different tissues of animals in cyanide detoxification.     

Study of mast cell and histamine contents of the pineal body

Summary The number and distribution of mast cells were studied in pineal bodies of 16 mammalian species. In cattle, goat, monkey and guinea-pig the cells occurred in large numbers; in man, sheep, marmoset and mouse there was a moderate amount, and they were almost absent in pineal bodies of horse, rat, pig, dog, hamster, fox, cat and rabbit.In 17 human pineal bodies, 11 of which were obtained from normal individuals, the mast cell population was 3 ± 0.5 mast cells per microscopical field (0.32 mm in diameter).The histamine content in pineal bodies of several mammals was also determined. The substance occurred in high amount in the pineal body of cattle, in moderate amount in that of goat, man and sheep, and could not be detected in horse, pig, rabbit and rat.In human and cattle pineal bodies the histamine content could be correlated to the mast cell population; in other species the same correlation was only suggested.In in vitro experiments octylamine released histamine from the pineal body of cattle, while compound 48/80 was ineffective.It is suggested that the hypotensive action of pineal extract reported by several authors might be due to the histamine content of this gland.This work was supported in part by funds received from the Conselho Nacional de Pesquisas of Brazil and The Rockefeller Foundation (Grant 61119). — Acknowledgement. For encouragement and criticism the authors wish to express their gratitude to Prof. Dr. W. T. Beraldo (Department of Physiology) and Prof. Dr. L. J. A. Di Dio (Department of Anatomy).Fellow of the Conselho Nacional de Pesquisas of Brazil.