Opisthorchis viverrini: ultrastructure and cytochemistry of the glycocalyx of the tegument

2000 ◽  
Vol 74 (1) ◽  
pp. 23-29 ◽  
Author(s):  
W. Apinhasmit ◽  
P. Sobhon ◽  
C. Tarasub ◽  
W. Mothong ◽  
P. Saitongdee ◽  
...  
Keyword(s):  
Sialic Acid ◽  
Binding Sites ◽  
Ricinus Communis ◽  
Ruthenium Red ◽  
Opisthorchis Viverrini ◽  
Binding Studies ◽  
Con A ◽  
Colloidal Iron ◽  
Dolichos Biflorus ◽  
Ulex Europaeus

AbstractThe ultrastructure and cytochemistry of the glycocalyx of the tegument of Opisthorchis viverrini during maturation from newly excysted juvenile to adult stages were investigated using colloidal iron, ruthenium red and lectin stainings. The results showed that the glycocalyx was intensely stained by the first two dyes, thus indicating the presence of relatively high amounts of negative charges. However, the thickness and intensity of the staining decreased during the fluke's maturation. Binding studies using lectin probes on the surface of adult parasites showed that binding sites for Canavaliaensiformis (Con A), Triticum vulgaris (WGA) and Ricinus communis I(RCA I) were present in relative large amounts on the glycocalyx of the adult tegument, whereas those for Dolichos biflorus (DBA) were relatively fewer in number, and those for Ulex europaeus I (UEA I) were absent. The binding patterns of Con A, WGA, RCA I and DBA were generally similar, and the reaction product was uniformly distributed over the dorsal and ventral surfaces of the parasite's body. These bindings, therefore, indicate the presence of D-mannose/D-glucose, N-acetyl-D-glucosamine/sialic acid, D-galactose and N-acetyl-D-galactosamine residues on the glycocalyx of the adult tegument.

scholarly journals Membrane structure and surface coat of Entamoeba histolytica. Topochemistry and dynamics of the cell surface: cap formation and microexudate.

1975 ◽  
Vol 64 (3) ◽  
pp. 538-550 ◽  
Author(s):  
P P Silva ◽  
A Martínez-Palomo ◽  
A Gonzalez-Robles
Keyword(s):  
Plasma Membrane ◽  
Cell Surface ◽  
Binding Sites ◽  
Membrane Structure ◽  
Ruthenium Red ◽  
Iron Binding ◽  
Con A ◽  
Colloidal Iron ◽  
Acidic Sites ◽  
Membrane Components

Treatment of living entamoeba histolytica cells with low concentrations of concanavalin A (con A) and peroxidase results in redistribution of the plasma membrane con A receptors to one pole of the cell where a morphologically distinct region--the uroid--is formed. Capping of con A receptors is not accompanied by parallel accumulation of ruthenium red-stainable components. In capped cells, the pattern of distribution of acidic sites ionized at pH 1.8 (labeled by colloidal iron) at the outer surface and of membrane particles (integral membrane components revealed by freeze-fracture) is not altered over the uroid region. Cytochemistry of substrate-attached microexudate located in regions adjacent to E. histolytica cells demonstrates the presence of con A binding sites and ruthenium red- and alcian blue-stainable components and the absent of colloidal iron binding sites. In a previous report we demonstrated that glycerol-induced aggregation of the plasma membrane particles is accompanied by a discontinuous distribution of colloidal iron binding sites, while con A receptors and acidic sites ionized at pH 4.0 remain uniformly distributed over the cell surface. Taken together, our experiments show that, in E. histolytica cells, peripheral membrane components may move independently of integral components and, also, that certain surface determinants may redistribute independently of others. These results point to the complexity of the membrane structure-cell surface relationship in E. histolytica plasma membranes relative to the membrane of the erythrocyte ghost where integral components (the membrane-intercalated particles) contain all antigens, receptors, and anionic sites labeled so far. We conclude that fluidity of integral membrane components (integral membrane fluidity) cannot be inferred from the demonstration of the mobility of surface components nor, conversely, can the fluidity of peripheral membrane components (peripheral membrane fluidity) be assumed from demonstration of the mobility of integral membrane components.

Analysis of Glycoproteins from Nasal Polyps and Papillomas by Affinity Chromatography

1984 ◽  
Vol 93 (1) ◽  
pp. 85-88 ◽  
Author(s):  
Katsunori Fukuda ◽  
Hirofumi Matsuyama ◽  
Kohzo Fukami ◽  
Masayuki Ozawa ◽  
Takashi Muramatsu ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Affinity Chromatography ◽  
Binding Sites ◽  
Nasal Polyps ◽  
Ricinus Communis ◽  
Sodium Dodecyl ◽  
Peanut Agglutinin ◽  
Con A ◽  
Dolichos Biflorus ◽  
Ricinus Communis Agglutinin

Glycoproteins were isolated from the particulate fraction of four nasal polyps and three nasal papillomas by affinity chromatography on lectins conjugated with agarose (Concanavalin A [Con A], wheat germ agglutinin [WGA], Ricinus communis agglutinin [RCA], peanut agglutinin [PNA], and Dolichos biflorus agglutinin [DBA]). The glycoprotein mixtures so isolated were then analyzed by sodium dodecyl sulfate gel electrophoresis. Glycoprotein profiles of nasal polyps were similar to each other, but were distinctively different from those of nasal papillomas. Binding sites for Con A, WGA, and RCA isolated from nasal papillomas contained intense bands with a molecular weight less than 15,000 daltons, which were absent in nasal polyps. The major component of PNA-binding sites of nasal polyps is of a molecular weight of 65,000 daltons, which was not detected in nasal papillomas.

Lectin Binding Studies on RNA Tumor Virus-Infected Cells

1975 ◽  
Vol 33 ◽  
pp. 326-327
Author(s):  
D. C. Hixson
Keyword(s):  
Binding Sites ◽  
Lectin Binding ◽  
Culture Conditions ◽  
3T3 Cells ◽  
Binding Studies ◽  
Con A ◽  
Microscopic Studies ◽  
Tumor Virus ◽  
Infected Cells ◽  
Cell Culture Conditions

The abilities of plant lectins to preferentially agglutinate malignant cells and to bind to specific monosaccharide or oligosaccharide sequences of glycoproteins and glycolipids make them a new and important biochemical probe for investigating alterations in plasma membrane structure which may result from malignant transformation. Electron and light microscopic studies have demonstrated clustered binding sites on surfaces of SV40-infected or tryp- sinized 3T3 cells when labeled with concanavalin A (con A). No clustering of con A binding sites was observed in normal 3T3 cells. It has been proposed that topological rearrangement of lectin binding sites into clusters enables con A to agglutinate SV40-infected or trypsinized 3T3 cells (1). However, observations by other investigators have not been consistent with this proposal (2) perhaps due to differences in reagents used, cell culture conditions, or labeling techniques. The present work was undertaken to study the lectin binding properties of normal and RNA tumor virus-infected cells and their associated viruses using lectins and ferritin-conjugated lectins of five different specificities.

Echinostoma caproni and E. trivolvis alter the binding of glycoconjugates in the intestinal mucosa of C3H mice as determined by lectin histochemistry

1993 ◽  
Vol 67 (3) ◽  
pp. 179-188 ◽  
Author(s):  
T. Fujino ◽  
B. Fried
Keyword(s):  
Ricinus Communis ◽  
Villous Atrophy ◽  
Lectin Binding ◽  
Goblet Cells ◽  
Lectin Histochemistry ◽  
Dolichos Biflorus ◽  
Echinostoma Caproni ◽  
Ulex Europaeus ◽  
C3h Mice ◽  
Small Intestines

AbstractMouse (C3H) mucosal glycoconjugates were examined in normal small intestines and intestines infected with Echinostoma caproni, or E. trivolvis using six different fluorescein-conjugated lectins: Triticum, vulgaris agglutinin (WGA), Ulex europaeus agglutinin I (UEA-I), Ricinus communis agglutinin I (RCA-I). Glycine max soybean agglutinin (SBA), Dolichos biflorus agglutinin (DBA), and Arachis hypogaeu peanut agglutinin (PNA). The expression of lectin-binding sites and the intensity of the binding of lectins in the mouse small intestines were changed by infection with the echinostomes. Specific differences in the reaction to glycoproteins were clearly observed between the mouse intestines infected with E. caproni and those infected with E. trivolvis. In E. caproni infection, binding of most of the lectins to the villi was remarkably reduced in accord with the villous atrophy and loss of goblet cells. In contrast, in E. trivolvis infection, the binding of WGA, RCA-I and DBA was reduced in the microvillar surfaces, but binding of UEA-I and SBA were unchanged compared to the control intestines. The lectin binding to goblet cells in E. trivolvis-infected mice mostly increased. These observations may reflect the marked increase in goblet cells and the less severe damage in the villi of E. trivolvis infection compared to E. caproni infection. Most of the glycoconjugates were slightly reduced in the hyperplastic crypts except for N-acetyl glucosamine. It is possible that glucose metabolism in the host intestines infected with E. trivolvis was activated. resulting in an increase in the rate of mucin synthesis as well as qualitative changes in mucus, thereby mediating the expulsion of the worms.

scholarly journals Ultrastructural localization of lectin-binding sites on the zonae pellucidae and plasma membranes of mammalian eggs.

1975 ◽  
Vol 66 (2) ◽  
pp. 263-274 ◽  
Author(s):  
G L Nicolson ◽  
R Yanagimachi ◽  
H Yanagimachi
Keyword(s):  
Binding Sites ◽  
Plasma Membranes ◽  
Ricinus Communis ◽  
Lectin Binding ◽  
Ultrastructural Localization ◽  
Con A ◽  
Plasma Membrane Receptor ◽  
Rat Eggs ◽  
Mammalian Eggs ◽  
Lectin Binding Sites

Receptors for Ricinus communis agglutinin I (RCAI), concanavalin A (Con A), and wheat germ agglutinin (WGA) were localized on the zonae pellucidae and plasma membranes of hamster, mouse, and rat eggs with ferritin-lectin conjugates. Intact eggs labeled with the ferritin conjugates showed dense concentrations of RCAI and WGA receptors in the outermost regions of their zonae pellucidae and sparse distributions of Con A receptors throughout the zonae. Ferritin-lectin labeling was specific, since inhibitory saccharides effectively blocked labeling. The asymmetric density of RCAI receptors across the zona was confirmed by ferritin-RCAI and fluorescein-RCAI labeling of mechanically isolated zonae pellucidae, indicating that the RCAI-binding sites are more densely distributed in the exterior zona regions. Plasma membranes of rodent eggs contained RCAI, WGA, and Con A receptors. These receptors were found to be more or less randomly distributed on surfaces of aldehyde-fixed eggs or on eggs labeled near 0 degrees C. However, eggs incubated at 25 degrees C showed aggregated WGA- and Con A-binding site distributions on their plasma membranes. This indicates that lectin-induced receptor redistribution occurs at this temperature. The possibility that plasma membrane receptor mobility is a requirement for sperm-egg fusion is discussed.

scholarly journals Lectin-binding sites on the plasma membranes of rabbit spermatozoa: Changes in surface receptors during epididymal maturation and after ejaculation

1977 ◽  
Vol 74 (3) ◽  
pp. 950-962 ◽  
Author(s):  
GL Nicolson ◽  
N Usui ◽  
R Yanagimachi ◽  
H Yanagimachi ◽  
Smith JR
Keyword(s):  
Binding Sites ◽  
Plasma Membranes ◽  
Ricinus Communis ◽  
Lectin Binding ◽  
Similar Degree ◽  
Con A ◽  
Lectin Receptors ◽  
Surface Receptors ◽  
Epididymal Spermatozoa ◽  
Lectin Binding Sites

Modifications in rabbit sperm plasma membranes during epididymal passage and after ejaculation were investigated by used of three lectins: concanavalin A (Con A); Ricinus communis I (RCA(I)); and wheat germ agglutinin (WGA). During sperm passage from caput to cauda epididymis, agglutination by WGA drastically decreased, and agglutination by RCA(I) slightly decreased, although agglutination by Con A remained approximately unchanged. After ejaculation, spermatozoa were agglutinated to a similar degree or slightly less by Con A, WGA, and RCA(I), compared to cauda epididymal spermatozoa. Ultrastructural examination of sperm lectin-binding sites with ferritin- lectin conjugates revealed differences in the densities of lectin receptors in various sperm regions, and changes in the same regions during epididymal passage and after ejaculation. Ferritin-RCA(I) showed abrupt changes in lectin site densities between acrosomal and postacrosomal regions of sperm heads. The relative amounts of ferritin-RCA(I) bound to heads of caput epididymal or ejaculated spermatozoa. Tail regions were labeled by ferritin RCA(I) almost equally on caput and cauda epididymal spermatozoa, but the middle-piece region of ejaculated spermatozoa was slightly more densely labeled than the principal-piece region, and these two regions on ejaculated spermatozoa were labeled less than on caput and cuada epididymal spermatozoa. Ferritin-WGA densely labeled the acrosomal region of caput epididymal spermatozoa, although labeling of cauda epidiymal spermatozoa was relatively sparse except in the apical area of the acrosomal region. Ejaculated spermatozoa bound only a few molecules of ferritin-WGA, even at the highest conjugate concentrations used. Caput epididymal, but not cauda epididymal or ejaculated spermatozoa, bound ferritin-WGA in the tail regions. Dramatic differences in labeling densities during epididymal passage and after ejaculation were not found with ferritin-Con A.

scholarly journals A cytochemical study of lectin receptors on isolated chick neural retina neurons in vitro

1982 ◽  
Vol 53 (1) ◽  
pp. 1-20
Author(s):  
J.A. Bee
Keyword(s):  
Sialic Acid ◽  
Cell Body ◽  
Growth Cone ◽  
Ricinus Communis ◽  
Lectin Binding ◽  
Neuronal Cell ◽  
Cytochemical Study ◽  
Lectin Receptors ◽  
Dolichos Biflorus ◽  
Dolichos Biflorus Agglutinin

The cell body, neurite and growth cone of isolated retinal neurons have been compared on the basis of their ability to bind a number of fluorescently labelled lectins, each possessing a unique carbohydrate specificity. The susceptibility of the respective binding patterns following pretreatment of these fixed cells with either neuraminidase or trypsin was also investigated. Neuronal cell bodies displayed the most intense binding of each lectin, with localization of limulin binding (specific for sialic acid) predominantly to the neurite hillock, the point on the cell body from which the neurite projects. Limulin binding was almost totally abolished by pretreatment with either neuraminidase or trypsin. In contrast to the cell body, limulin binding to the neurite or growth cone was not detected. These regions of the cell apparently possessed sialic acid, however, since pretreatment with neuraminidase reduced wheat germ agglutinin binding (to N-acetylglucosamine) and markedly enhanced Dolichos biflorus agglutinin binding (to N-acetylgalactosamine) to both the neurite and growth cone. The initially low binding of Dolichos biflorus agglutinin to the neurite and growth cone was slightly enhanced by pretreatment with trypsin. Uniformly low levels of binding of either Ricinus communis agglutinin 60 (galactose, N-acetylgalactosamine) or R. communis agglutinin 120 (galactose) was observed over the entire neuron. R. communis agglutinin 120 binding was not enhanced by pretreatment with neuraminidase. Receptors for either concanavalin A (mannose, glucose) or Ulex europaeus agglutinin I (fucose) were abundant over the entire nerve cell with the former exhibiting more marked trypsin sensitivity. From these data, it is apparent that the repertoire of lectin binding sites of the neurite and growth cone of these differentiating nerve cells differs markedly from that of the cell body, which itself demonstrates some degree of regionalization.

scholarly journals Localization of carbohydrate components in rat colon with fluoresceinated lectins.

1978 ◽  
Vol 26 (6) ◽  
pp. 452-458 ◽  
Author(s):  
E Essner ◽  
J Schreiber ◽  
R A Griewski
Keyword(s):  
Epithelial Cells ◽  
Concanavalin A ◽  
Wheat Germ ◽  
Ricinus Communis ◽  
Rat Colon ◽  
Dolichos Biflorus ◽  
Ulex Europaeus ◽  
Descending Colon ◽  
Carbohydrate Components ◽  
Cryostat Sections

Cryostat sections of rat descending colon were studied by fluorescence microscopy after exposure to conjugates of fluorescein isothicoyanate with lectins from Glycine max (soybean), Triticum vulgaris (wheat germ), Ricinus communis (castor bean), Ulex europaeus, (gorse), Dolichos biflorus (horse gram) and Canavalia ensiformis (concanavalin A) (Jack bean). No two lectins showed identical patterns of fluorescence. FITC-conjugates of soybean and D. biflorus lectins reacted strongly with the mucus present in the crypt lumens and with the surface (as well as cytoplasm) of the epithelial cells suggesting that these sites are rich in terminal, non-reducing, N-acetylgalactosamine residues. Wheat germ, R. communis, U. europaeus and concanavalin A-FITC conjugates did not stain mucus but showed fluorescence in the cytoplasm of absorptive cells as well as in the lamina propria and submucosa. The FITC-R. communis conjugate also reacted with structures in the apical portion of epithelial cells that may correspond to the Golgi apparatus.

scholarly journals Binding studies of a sialic acid-specific lectin from the horseshoe crab Carcinoscorpius rotunda cauda with various sialoglycoproteins

1982 ◽  
Vol 203 (1) ◽  
pp. 253-261 ◽  
Author(s):  
S Mohan ◽  
D Thambi Dorai ◽  
S Srimal ◽  
B K Bachhawat
Keyword(s):  
Sialic Acid ◽  
Binding Sites ◽  
Horseshoe Crab ◽  
Binding Studies ◽  
Sialic Acid Content ◽  
Precipitation Inhibition ◽  
Bivalent Metal Ions ◽  
Acidic Sugars ◽  
Inhibition Studies

Interaction of the sialic acid-specific lectin carcinoscorpin with various sialoglycoproteins was studied by using radioiodinated lectin. The binding of carcinoscorpin was dependent not only on sialic acid content but also on the type of glycosidic linkage and form (branched or linear) of the carbohydrate chains. Carcinoscorpin has different classes of binding sites, and binding follows a phenomenon of positive co-operativity. The effect of Ca2+ concentration on the binding was studied, and the optimal concentration was found to be 0.02 M. Effect of pH, temperature and other bivalent metal ions are also reported. From haemagglutination- and precipitation-inhibition studies, it was concluded that carcinoscorpin has multispecificity towards acidic sugars, and its relation to the biological role of the lectin in the horseshoe crab is discussed.

Sign in / Sign up

Export Citation Format

Share Document