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 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

1979 ◽  
Vol 80 (1) ◽  
pp. 69-76 ◽  
Author(s):  
M Maylie-Pfenninger ◽  
JD Jamieson
Keyword(s):  
Cell Surface ◽  
General Procedure ◽  
Acinar Cells ◽  
Pancreatic Acinar Cells ◽  
Affinity Constant ◽  
Affinity Column ◽  
Gel Chromatography ◽  
Binding Studies ◽  
Binding Properties ◽  
Pancreatic Cells

We describe here a simple, general procedure for the purification of a variety of lectins, and for the preparation of lectin-ferritin conjugates of defined molar composition and binding properties to be used as probes for cell surface saccharides. The technique uses a "universal" affinity column for lectins and their conjugates, which consists of hog sulfated gastric mucin glycopeptides covalently coupled to agarose. The procedure involes: (a) purification of lectins by chromatography of aqueous extracts of seeds or other lectin-containing fluids over the affinity column, followed by desorption of the desired lectin with its hapten suge; (b) iodination of the lectin to serve as a marker during subsequent steps; (c) conjugation of lectin to ferritin with glutaraldehyde; (d) collection of active lectin-ferritin conjugates by affinity chromatography; and (e) separation of monomeric lectin-ferritin conjugates from larger aggregates and unconjugated lectin by gel chromatography. Based on radioactivity and absorbancy at 310 nm for lectin and ferritin, respectively, the conjugates consist of one to two molecules of lectin per ferrritin molecule. Binding studies of native lectins and their ferritin conjugates to dispersed pancreatic acinar cells showed that the conjugation procedure does not significantly alter either the affinity constant of the lectin for its receptor on the cell surface or the number of sites detected.

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.

Receptor for calcitonin gene-related peptide: binding to exocrine pancreas mediates biological actions

1985 ◽  
Vol 249 (1) ◽  
pp. G147-G151 ◽  
Author(s):  
H. Seifert ◽  
P. Sawchenko ◽  
J. Chesnut ◽  
J. Rivier ◽  
W. Vale ◽  
...  
Keyword(s):  
Guinea Pig ◽  
Islets Of Langerhans ◽  
Exocrine Pancreas ◽  
Acinar Cells ◽  
Calcitonin Gene Related Peptide ◽  
Pancreatic Acinar Cells ◽  
Binding Studies ◽  
Amylase Release ◽  
Related Peptide ◽  
Calcitonin Gene

In the present study we demonstrate by immunohistochemical techniques that calcitonin gene-related peptide (CGRP) is present in nerve terminals in the islets of Langerhans. Furthermore, binding studies with 125I-CGRP indicate that dispersed acini from guinea pig pancreas contain a single class of high-affinity binding sites for CGRP with an apparent dissociation constant of 18 nM. Vasoactive intestinal peptide (VIP), rat growth hormone-releasing factor (rGRF), cholecystokinin octapeptide (CCK-OP), and bombesin do not interact with these receptors. Interaction of CGRP with these receptors leads to release of amylase from the acinar cells. Amylase release is half maximal at 0.3 nM CGRP and maximal at 3 nM CGRP. Maximal amylase release with CGRP is one-third of that observed with VIP. CGRP-induced amylase release is dependent on theophylline in the incubation medium. CGRP potentiates the amylase release stimulated by bombesin and CCK-OP but has no effect on amylase release stimulated by VIP, rGRF, and natural glucagon. CGRP stimulates a 25% increase in basal cellular cAMP. These results indicate that guinea pig pancreatic acinar cells contain a novel receptor for CGRP and that interaction of CGRP with this receptor leads to digestive enzyme secretion through a cAMP-mediated pathway. The presence of CGRP in the islets of Langerhans suggests a pathway for CGRP to reach the exocrine pancreas through an insuloacinar portal system.

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.

Biochemical characterization of some raptor trypanosomes. I. In vitro cultivation and lectin-binding studies

1986 ◽  
Vol 64 (1) ◽  
pp. 189-194 ◽  
Author(s):  
Carl E. Kirkpatrick ◽  
Cynthia A. Terway-Thompson
Keyword(s):  
Biochemical Characterization ◽  
Ricinus Communis ◽  
Lectin Binding ◽  
The Other ◽  
In Vitro Cultivation ◽  
Type I ◽  
Binding Studies ◽  
Binding Characteristics ◽  
American Kestrels

Nine trypanosome strains from five species of raptors were cultivated in vitro in a monophasic medium. Two morphologically distinct trypanosomes were observed in culture: those from American kestrels (Falco sparverius) were smaller than the other strains. The two kestrel (KT) trypanosome strains showed in vitro growth kinetics that differed from the larger trypanosomes, and the KT strains, unlike the others, required hemin in the medium for growth. The effectiveness of eight plant lectins to induce the agglutination of cultured trypanosomes was studied as a means of differentiating the various strains. It was found that lectins from Lens culinaris and Ricinus communis (type I) were particularly effective in distinguishing the KT strains from the other raptor trypanosome strains. Based on the results of experiments in which lectin-mediated trypanosome agglutination was inhibited by the addition of various monosaccharides, it is concluded that all of the avian trypanosomes studied express surface methyl α-D-mannoside, D(+)-galactose, and (or) α-lactose. Only the relatively large raptor trypanosome isolates expressed N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, and α-L(−)-fucose on their surfaces. The differences in lectin-binding characteristics between the two morphologic types of raptor trypanosome were as great as those among each of the avian trypanosomes and the mammalian trypanosomatids Leishmania chagasi and Trypanosoma rhodesiense.

Loss Of Platelet Membrane Glycoproteins Or Terminal Sialic Acid Detected By Fitc-Lectins

1981 ◽  
Author(s):  
L McGregor ◽  
J L McGregor ◽  
K J Clemetson ◽  
M Dechavanne ◽  
E F Lüscher
Keyword(s):  
Sialic Acid ◽  
Ricinus Communis ◽  
Lectin Binding ◽  
Human Platelets ◽  
Membrane Glycoproteins ◽  
Binding Studies ◽  
Platelet Surface ◽  
Terminal Sialic Acid

Pre-thrombic conditions in certain individuals resulting from enhanced platelet-vessel wall or platelet-platelet interactions are perhaps characterized by a reduction in certain membrane glycoproteins or loss of terminal sialic acid. In order to investigate if such changes are detectable, the binding of FITC-lectins to human platelets treated under in vitro conditions with certain proteases to mimic possible in vivo changes occuring on the platelet surface, has been examined. Human platelets were isolated, washed and either treated with neuraminidase (10 U) or plasmin (1 CU) before fixing with formaldehyde. Binding studies were performed by the method of Monsigny et al. using FITC labelled wheat germ agglutinin (WGA), Lens culinaris lectin (LCL), Ricinus communis agglutinin (RCA) and concanavalin A (ConA). The number of lectin-binding sites (n) and the dissociation constant (Kd) were obtained by Steck and Wallach reciprocal plots. After neuraminidase or plasmin treatment n was reduced but Kd remained approximately the same with WGA. FITC-RCA-60 gave a slight fluorescence with untreated and very strong fluorescence with neuraminidase treated platelets. Platelet glycoproteins separated by 2-dimensional gel electrophoresis were identified by binding of fluorescent lectins. Plasmin decreased the intensity of GP Ib and IIb and removed Ia completely. Neuraminidase decreased the labelling of Ib by WGA. These techniques show promise as methods of detecting pre-thrombotic conditions.

Intermediate Cells of the Pancreas

1973 ◽  
Vol 13 (1) ◽  
pp. 297-315
Author(s):  
R. N. MELMED ◽  
CAROL J. BENITEZ ◽  
S. J. HOLT
Keyword(s):  
Endocrine Cells ◽  
Experimental Diabetes ◽  
Acinar Cells ◽  
Cell Type ◽  
Β Cells ◽  
Β Cell ◽  
Rat Pancreas ◽  
Pancreatic Cells ◽  
Normal Rat ◽  
Intermediate Cells

In addition to causing necrosis of β-cells, diabetogenic doses of alloxan or streptozotocin cause selective autophagy and destruction of β-granules in intermediate cells in the rat pancreas, thus providing evidence that these granules and those in the β-cells of the islet are identical. In acinar-β cells the autophagic vacuoles also contain small mitochondria similar to those that occur in islet endocrine cells. Apart from these effects, the cells remain structurally intact. These observations suggest that alloxan exerts a direct effect on the β-components of the β-granule-containing intermediate cells. The destruction of the β-granules in the acinar-β cell is sometimes accompanied by the appearance of α-granules in the same cells, thus presumably reflecting the increased demand for glucagon that occurs in experimental diabetes. Although intermediate cells containing α-granules are very uncommon in the normal rat pancreas, acinar-α and α-acinar cells occurred much more frequently after alloxan treatment. The presence of intermediate cells of the acinar-β type in normal rats and the observed change of acinar-β to acinar-α cells, or from α- to α-acinar cells after alloxan, indicates that some pancreatic cells possess a variable functional potentiality. The appearance of these cells after alloxan presumably reflects the altered metabolic state of the animal, and is not a manifestation of the transformation of one cell type into another.

scholarly journals Development of cell surface saccharides on embryonic pancreatic cells

1980 ◽  
Vol 86 (1) ◽  
pp. 96-103 ◽  
Author(s):  
MF Maylie-Pfenninger ◽  
JD Jamieson
Keyword(s):  
Cell Surface ◽  
Binding Sites ◽  
Ricinus Communis ◽  
Adult Life ◽  
Acinar Cells ◽  
In Utero ◽  
Soybean Agglutinin ◽  
Early Postpartum ◽  
Adult Cell ◽  
Centroacinar Cells

Using a battery of seven lectin-ferritin conjugates as probes for cell surface glycoconjugates, we have studied the pattern of plasmalemmal differentiation of cells in the embryonic rat pancreas from day 15 in utero to the early postpartum stage. Our results indicate that differentiation of plasmalemmal glycoconjugates on acinar, endocrine, and centroacinar cells is temporally correlated with development and is unique for each cell type, as indicated by lectin-ferritin binding. Specifically, (a) expression of adult cell surface saccharide phenotype can be detected on presumptive acinar cells as early as 15 d in utero, as indicated by soybean agglutinin binding, and precedes development of intracellular organelles characteristic of mature acinar cells; (b) maturation of the plasmalemma of acinar cells is reached after intracellular cytodifferentiation is completed, as indicated by appearance of Con A and fucoselectin binding sites only at day 19 of development; conversely, maturation of the endocrine cell plasmalemma is accompanied by "loss" (masking) of ricinus communis II agglutinin receptors; and (c) binding sites for fucose lectins and for soybean agglutinin are absent on endocrine and centroacinar cells at all stages examined. We conclude that acinar, centroacinar, and endocrine cells develop from a common progenitor cell(s) whose plasmalemmal carbohydrate composition resembles most closely that of the adult centroacinar cell. Finally, appearance of acinar lumina beginning at approximately 17 d in utero is accompanied by differenetiation of apical and basolateral plasmalemmal domains of epithelial cells, as indicated by enhanced binding of several lectin-ferritin conjugates to the apical plasmalemmal, a pattern that persists from this stage through adult life.

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