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 Membrane glycoproteins of the nerve growth cone: diversity and growth regulation of oligosaccharides.

1986 ◽  
Vol 103 (4) ◽  
pp. 1369-1382 ◽  
Author(s):  
L M Greenberger ◽  
K H Pfenninger
Keyword(s):  
Sialic Acid ◽  
Growth Cone ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Growth Regulation ◽  
Binding Proteins ◽  
Ricinus Communis ◽  
Lectin Binding ◽  
Adult Brain ◽  
Two Dimensional

A subcellular fraction prepared from fetal rat brain and enriched in growth cone membranes is analyzed for its lectin-binding proteins. Growth-associated glycoproteins are identified by comparing the growth cone glycoproteins with those of synaptosomes. Protein was resolved in one- or two-dimensional gels, electroblotted, and blots probed with radioiodinated concanavalin A, wheat germ agglutinin, and Ricinus communis agglutinins I and II. In one-dimensional gels, each lectin recognizes approximately 20 polypeptides (with substantial overlap) most of which migrate diffusely and have relatively high molecular masses (range 30-200 kD). The seven major Coomassie-staining proteins of the membrane fraction (34-52 kD) are not the major lectin-binding proteins. In two-dimensional gels, the lectin-binding proteins are either streaked across the pH gradient or exist as multiple spots, indicating broad charge heterogeneity. Seven wheat germ agglutinin- and Ricinus communis agglutinin II-binding glycoproteins are present in greater abundance in growth cone fractions compared with synaptosomes. Most notably, an acidic, sialic acid-rich protein (27-30 kD, pI 4.0; termed gp27-30) is most abundant at postnatal day 4, but absent from adult brain. The protein's very acidic isoelectric point is due, at least in part, to its high sialic acid content. Growth regulation of specific protein-linked oligosaccharides suggests that they play a special role in growth cone function. In addition, the great diversity of growth cone glycoproteins from whole brain suggests glycoprotein heterogeneity among growth cones from different neuron types.

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 Immunogold study on lectin binding in the porcine zona pellucida and granulosa cells

10.4081/846 ◽  
2009 ◽  
Vol 47 (4) ◽  
pp. 353 ◽  
Author(s):  
F Parillo ◽  
C Dall’Aglio ◽  
A Verini Supplizi ◽  
P Ceccarelli ◽  
AM Gargiulo
Keyword(s):  
Sialic Acid ◽  
Horseradish Peroxidase ◽  
Granulosa Cells ◽  
Zona Pellucida ◽  
Lectin Binding ◽  
Protein A ◽  
Ultrastructural Localization ◽  
Binding Pattern ◽  
Con A ◽  
Lectin Receptors

An ultrastructural localization of lectin receptors on the zona pellucida (ZP) of porcine antral oocytes and on the granulosa cells was performed using a panel of horseradish peroxidase- labelled lectins in conjunction with antiperoxidase antibody and protein A-gold. In some cases, lectin incubation was preceded by sialidase digestion. WGA-, Con-A-, UEA-I-, RCA-I-, PNA- and SBA-reactive sites were distributed differently in the porcine ZP. Sialidase digestion increased the positivity obtained with RCA-I and it was necessary to promote PNA and SBA reactivity. These results indicated that the ZP contained N-acetylglucosamine, a-mannose, a- fucose, b-Gal-(1-4)GlcNAc, b-Gal- (1-3)GalNAc, b-GalNAc and sialic acid residues. We also observed the presence of vesicles in both the ooplasm and granulosa cells, showing a similar lectin binding pattern to that of the ZP, thus suggesting that the oocyte and granulosa cells are the site of synthesis of ZP glucidic determinants.

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 Distribution of cell surface saccharides on pancreatic cells. II. Lectin-labeling patterns on mature guinea pig and rat pancreatic cells.

1979 ◽  
Vol 80 (1) ◽  
pp. 77-95 ◽  
Author(s):  
M F Maylié-Pfenninger ◽  
J D Jamieson
Keyword(s):  
Guinea Pig ◽  
Endocrine Cells ◽  
Ricinus Communis ◽  
Lectin Binding ◽  
Acinar Cells ◽  
Binding Studies ◽  
Lectin Receptors ◽  
Pancreatic Cells ◽  
Receptor Sites ◽  
Centroacinar Cells

The surface saccharide composition of collagenase-dispersed pancreatic cells from adult guinea pig and rat glands was examined by using eight lectins and their ferritin conjugates: Concanavalin A (ConA); Lens culinaris (LCL); Lotus tetragonolobus (LTL); Ricinus communis agglutinins I and II (RCA I, RCA II); Soybean agglutinin (SBA); Ulex europeus lectin (UEL); and wheat germ agglutinin (WGA). Binding studies of iodinated lectins and lectin-ferritin conjugates both revealed one population of saturable, high-affinity receptor sites on the total cell population (approximately 95% acinar cells). Electron microscopy, however, revealed differences in lectin-ferritin binding to the plasmalemma of acinar, centroacinar, and endocrine cells. Whereas acinar cells bound heavily all lectin conjugates, endocrine and centroacinar cells were densely labeled only by ConA, LCL, WGA, and RCA I, and possessed few receptors for LTL, UEL, and SBA. Endocrine and centroacinar cells could be differentiated from each other by using RCA II, which binds to centroacinar cells but not to endocrine cells. Some RCA II receptors appeared to be glycolipids because they were extracted by ethanol and chloroform-methanol in contrast to WGA receptors which resisted solvent treatment but were partly removed by papain digestion. RCA I receptors were affected by neither treatment. The apparent absence of receptors for SBA on endocrine and centroacinar cells, and for RCA II on endocrine cells, was reversed by neuraminidase digestion, which suggested masking of lectin receptors by sialic acid. The absence of LTL and UEL receptors on endocrine and centroacinar cells was not reversed by neuraminidase. We suggest that the differential lectin-binding patterns observed on acinar, centroacinar, and endocrine cells from the adult pancreas surface-carbohydrate-developmental programs expressed during morphogenesis and cytodifferentiation of the gland.

scholarly journals Hepatocyte cell surface polarity as demonstrated by lectin binding.

1988 ◽  
Vol 36 (12) ◽  
pp. 1561-1571 ◽  
Author(s):  
P N McMillan ◽  
D C Hixson ◽  
K A Hevey ◽  
S Naik ◽  
H O Jauregui
Keyword(s):  
Cell Surface ◽  
Binding Sites ◽  
Ricinus Communis ◽  
Lectin Binding ◽  
Separation Procedure ◽  
Lectin Receptors ◽  
Mechanical Methods ◽  
Dissociated Cells ◽  
Lectin Binding Sites

We performed an investigation at the ultrastructural level of the differential distribution of lectin-binding sites among sinusoidal, lateral, and bile canalicular domains of adult rat hepatocytes. Lectin binding to hepatocyte glycocalices was studied in situ or after cellular dissociation by enzymatic (collagenase), chemical (EDTA), and mechanical methods, as well as during cell culture. Using thirteen biotinylated lectins and an avidin-biotin-peroxidase complex (ABC), we have identified lectin-binding sites that are predominantly localized in the bile canalicular [Ricinus communis agglutinin (RCA)] or sinusoidal [Phaseolus vulgaris (PHA)] domains in situ and in mechanically dissociated cells. Lens culinaris (LCA) staining was prominent on sinusoidal surfaces, slight along lateral surfaces, and completely absent in the bile canalicular domain. Concanavalin A (ConA) was unique in binding equally to all domains. Triticum vulgaris [wheat germ agglutinin (WGA)] was also bound to all domains, but most intensely to the bile canalicular region. Cells dissociated via collagenase or EDTA treatment exhibited a spherical morphology characterized by many surface microvilli and absence of morphological domains. Lectin binding to dissociated cells was uniformly distributed over the entire cell surface, suggesting a redistribution of lectin receptors that was independent of the separation procedure. Hepatocytes in culture exhibited a partial restoration of morphological domains, but lectin binding polarity was not re-established.

Subdomains of the rough endoplasmic reticulum in colon goblet cells of the rat: lectin-cytochemical characterization.

1992 ◽  
Vol 40 (7) ◽  
pp. 919-930 ◽  
Author(s):  
A Ellinger ◽  
M Pavelka
Keyword(s):  
Endoplasmic Reticulum ◽  
Sialic Acid ◽  
Golgi Apparatus ◽  
Rough Endoplasmic Reticulum ◽  
Ricinus Communis ◽  
Secretory Granules ◽  
Lectin Binding ◽  
Helix Pomatia ◽  
Goblet Cells ◽  
Rat Colon

Using lectin binding, we characterized subdomains of the rough endoplasmic reticulum (rER) in goblet cells of the rat colon. In this cell type, special rER regions can be differentiated on the basis of their content of low electron density and dilated cisternal spaces in conventional transmission electron microscopic preparations. The fine fibrillar content of these cisternal regions demonstrated high-affinity binding with lectins from wheat germ, Helix pomatia, Griffonia simplicifolia I-A4 and -B4, and Ricinus communis I, although not with the sialic acid-specific Limax flavus lectin and the fucose-binding Ulex europaeus I lectin. Sugar-inhibitory experiments indicated that glycoconjugates packed within these regions bound the lectins with higher affinity than molecules present in the Golgi apparatus and secretory granules. Furthermore, the lectin binding patterns of the rER subdomains differed from those of the Golgi apparatus and mucin granules: the terminal sugar residues sialic acid and fucose were demonstrable in the Golgi apparatus and mucin granules and were absent from the rER, while galactose-recognizing lectins bound intensely at these rER regions, weakly to Golgi elements, and were almost absent from mucin granules.

scholarly journals A comparison of lectin binding in rat and human peripheral nerve.

1986 ◽  
Vol 34 (11) ◽  
pp. 1487-1493 ◽  
Author(s):  
A K Gulati ◽  
A A Zalewski ◽  
K B Sharma ◽  
D Ogrowsky ◽  
G S Sohal
Keyword(s):  
Peripheral Nerves ◽  
Ricinus Communis ◽  
Lectin Binding ◽  
Nerve Degeneration ◽  
Binding Affinities ◽  
Light Microscopic Level ◽  
Dolichos Biflorus ◽  
Ulex Europaeus ◽  
Maclura Pomifera ◽  
Subsequent Regeneration

Eleven different fluorescein- or peroxidase-conjugated lectins with different sugar-binding affinities were employed to analyze and compare glycoconjugates of rat and human peripheral nerves at the light microscopic level. A majority of lectins showed a distinct binding pattern in different structures of the nerve. Lectin binding was similar but not identical in rat and human nerves. Limulus polyhemus agglutinin did not stain any structures in rat or human nerves. In both species, all other lectins bound to the perineurium. Perineurial staining was intense with Canavalia ensiformis (Con A), Triticum vulgaris (WGA), Maclura pomifera (MPA); moderate with Glycine max (SBA), Griffonia simplicifolia-I (GS-I) and GS-II; weak with Ulex europaeus (UEA), Dolichos biflorus (DBA), and Ricinus communis (RCA). In the endoneurium of both species, ConA staining was intense, MPA and WGA moderate, SBA, GS-II, PNA, and RCA weak, and UEA and DBA absent. Interestingly, GS-I stained rat but not human endoneurium. Most lectins bound to blood vessels. GS-I bound to rat but not human, whereas UEA bound to human but not rat vessels. The results show that lectins can be used to reveal heterogeneity in sugar residues of glycoconjugates within neural and vascular components of nerves. They may therefore be potentially useful in detecting changes in glycoconjugates during nerve degeneration and subsequent regeneration after trauma or in pathological states.

scholarly journals Detection of sialic acid residues in the axonal reticulum of rat superior cervical ganglion cells by lectin-gold cytochemistry.

1990 ◽  
Vol 38 (10) ◽  
pp. 1445-1449 ◽  
Author(s):  
J R Quatacker ◽  
W G Annaert ◽  
W P De Potter
Keyword(s):  
Sialic Acid ◽  
Superior Cervical Ganglion ◽  
Plasma Membranes ◽  
Ganglion Cells ◽  
Ricinus Communis ◽  
Neuronal Cell ◽  
Thin Sections ◽  
Trans Golgi Network ◽  
Cervical Ganglion ◽  
Golgi Network

Highly glycosylated compounds have been demonstrated in the axonal reticulum elements of the superior cervical ganglion cells of the rat, and this is considered to suggest a connection of the reticulum with the trans Golgi side. In the present study, the axonal reticulum and the Golgi elements were further characterized by post-embedding methods of lectin-gold cytochemistry to determine their carbohydrate residues and to see, more specifically, if sialic acid residues could be detected in the axonal reticulum elements. Therefore, the affinity of neuronal cell structures for Limax flavus agglutinin (LFA), wheat germ agglutinin (WGA), and Ricinus communis agglutinin I (RCA-I) was tested in ultra-thin sections of glycolmethacrylate-embedded material, counterstained with phosphotungstic acid (PTA) at low pH. The trans Golgi network, the Golgi-associated axonal reticulum, the reticulum within axons, the large dense-cored vesicles, and the plasma membranes were reactive for all three lectins used. We conclude that the axonal reticulum elements carry sialic acid residues, relating them to the trans Golgi network. The present results support the concept that the axonal reticulum is an extension of the trans network of the Golgi apparatus specialized for neurosecretion.

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