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Differential distribution of lectin-binding glycoconjugates in the secretory granules of hamster oviductal ampulla during the estrous cycle: a quantitative cytochemical analysis.

High resolution lectin-gold cytochemistry was used to quantitatively analyze the distribution of glycoconjugates in the hamster oviductal ampulla during the five stages of the estrous cycle. Lectins binding to N-acetyl-D-galactosamine-, D-galactose-, and sialic acid-associated glycoconjugates in the secretory granules of ampullary epithelial secretory cells showed staining of equal intensity throughout the five different stages of the estrous cycle. In contrast, the labeling intensity of glycoconjugates which contain N-acetylglucosamine as terminal sugar residues reached its maximum around the time of ovulation, i.e., at proestrus. Glycoconjugates which carry fucose and mannose as terminal sugar residues appeared to be totally absent from the secretory granules of the oviductal ampulla during the estrous cycle. Together, electron microscopic observations combined with quantitative results indicate that N-acetyl-D-galactosamine-, D-galactose-, and sialic acid-associated glycoconjugates may be secreted into the ampullary lumen irrespective of the stage of the estrous cycle, whereas the secretion of certain N-acetylglucosamine-associated glycoconjugates is stage specific and reaches its peak at the time of ovulation. These findings suggest that, at the time of ovulation, the ampullary epithelium changes its secretory activity and contributes its secretory products to the zona pellucida of oocytes freshly released from the ovary.

Animals↗

Effects of benzimidazole anthelmintics as microtubule-active drugs on the synthesis and transport of surface glycoconjugates in Hymenolepis microstoma, Echinostoma caproni, and Schistosoma mansoni.

Hymenolepis microstoma (Cestoda), Echinostoma caproni, and Schistosoma mansoni (Digenea) were exposed to benzimidazoles to determine the influence of the drugs on the secretion of glycoconjugates that protect the worms' surface. Worms were obtained from mice treated with mebendazole or albendazole, and the glycoconjugates were localized in the parasite tissues by cytochemistry using lectin-gold conjugates. Events leading to the death of H. microstoma and E. caproni extended over a medication period for at least 2-3 days, and the following interrelated phases were discernible. Upon depolymerization of the microtubules the tegumentary cytons continued to synthesize glycoconjugates for up to about 24 h. Vesicles containing the glycans accumulated in the cytons, but their microtubule-based transport to the distal tegument was inhibited. At about 1 day the Golgi complex became fragmented and the production of glycans sharply declined. As a consequence of this and an ongoing turnover of the surface coat the contents of glycoconjugates in the distal tegument decreased. Similar effects were produced by vinblastine and colchicine in vitro. In contrast, benzimidazole treatment of S. mansoni, which is reportedly inefficacious, did not alter the replenishment of the surface glycoconjugates. Diminution of the coating with glycoconjugates of the surface of drug-sensitive species constitutes a secondary effect of benzimidazoles that might, synergistically with immune mechanisms of the host, enhance the expulsion of the worms.

Albendazole↗

Released glycoconjugate of indigenous Leishmania major enhances survival of a foreign L. major in Phlebotomus papatasi.

The effect of Leishmania glycoconjugate in the vector was investigated using Phlebotomus papatasi artificially infected with a Leishmania major strain that this vector does not transmit in nature. Glycoconjugate of the vector-specific strain of L. major was added to the infective meals of some fly groups and the success of infections with or without this substance was compared 4 d later. In the absence of glycoconjugate the parasites survived in 15.6% of the flies, while the addition of 0.5 mg/ml and 2 mg/ml raised the rate of infection to 34.3% and 63.0% respectively. Undigested blood was found in the stomach of 7.8% of the infected flies, whereas following meals with parasites and 2 mg/ml glycoconjugate it was present in 37% of the flies. The results demonstrated that this glycoconjugate increased the viability of the parasites in the unsuitable vector and delayed digestion of the infective meals. In a parallel experiment the glycoconjugate of L. donovani did not affect the survival of the parasites.

Animals↗

Membrane glycoconjugates as potential mediators of alcohol effects.

1. Membrane glycoconjugates include glycoproteins and glycolipids that have many important functions in a wide variety of tissues, especially brain. 2. Alcohol's ability to fluidize and swell plasma membranes could be expected to alter the orientation and conformation of the embedded glycoconjugates. 3. Both kinds of glycoconjugates can contain terminal moieties of sialic acid, which has been shown to be decreased by single doses of alcohol. Chronic exposure to alcohol may have no effect on sialic acid, except in very young animals. 4. Glycolipids containing sialic acid (gangliosides) are also decreased by acute doses of alcohol, but chronic alcohol has little effect. Thus, gangliosides may have a role in the development and expression of tolerance. 5. Glycoproteins containing sialic acid may also be involved in alcohol action, but there has been less research in this area. 6. Alcohol-induced disruptions in membrane glycoconjugates could affect the important cellular functions that glycoconjugates have, and thus research on alcohol effects on glycoconjugates could lead to important discoveries of diagnostic and therapeutic value for alcohol abuse and alcoholism.

Alcoholic Intoxication↗

Modulatory efficacy of green tea polyphenols on glycoconjugates and immunological markers in 4-Nitroquinoline 1-oxide-induced oral carcinogenesis-A therapeutic approach.

Green tea polyphenols (GTP) has been used as a chemopreventive agent world wide against chemically induced cancer. The present study is aimed to understand the therapeutic action of GTP on glycoconjugates and immunological markers in 4-Nitroquinoline 1-oxide (4-NQO)-induced oral cancer over a period of 30 days at 200mg/kg, p.o., Oral cancer was induced by painting 4-NQO for 8 weeks followed by administration of GTP after 22 weeks, for 30 days. Glycoconjugates such as hexose, hexosamine, sialicacid, fucose and mucoprotein were analysed. Expression of glycoconjugates was examined through histology and SDS-PAGE. Immunological markers such as circulating immune complex and mast cell density were studied. Oral cancer-induced animals showed a significant increase in levels of glycoconjugates and its expression, similar to that observed for immunological markers. Treatment with GTP altered the expression of glycoconjugates as well as immunological markers. The results suggest that GTP modulates both the expression of glycoconjugates and immunological markers resulting in regression of oral cancer.

4-Nitroquinoline-1-oxide↗

Analysis of glycoconjugates in patients with oral squamous cell carcinoma.

BACKGROUND: Our aim was to examine the levels of glycoconjugates in plasma, erythrocyte membranes and buccal mucosa of healthy subjects and oral cancer patients. SUBJECTS AND METHODS: This study was conducted on 48 adult male oral cancer patients with various clinical stages (stage II to stage IV; 16 of each) and 16 disease-free healthy subjects who underwent surgical removal of impacted teeth or vestibuloplasty without inflammation. RESULTS: The plasma and tumor tissues glycoconjugates levels were significantly increased, whereas the erythrocyte membranes glycoconjugates were significantly decreased in oral cancer patients as compared to healthy subjects. The levels of glycoconjugates were gradually increased from stage II to stage IV in plasma and tumor tissues and decreased in erythrocyte membranes from stage II to stage IV of oral cancer patients. CONCLUSION: The increased plasma glycoconjugates can be due to the expense of erythrocyte membrane glycoconjugates or tumor tissue itself.

Adult↗

Characterization of glycoconjugates of guinea pig seminal vesicle by lectin histochemistry.

In the present study, the expression of glycoconjugates in the guinea pig seminal vesicle was localized and partially characterized by lectin histochemistry using a battery of 30 different lectins specific for different carbohydrate residues. The results indicate that the glandular epithelium of the guinea pig seminal vesicle exhibits complex glycoconjugates rich in Man, beta-GlcNAc, beta-Gal, alpha/beta-GalNAc, Fuc and complex NeuAc(alpha2,6)Gal/GalNAc residues, as shown by its positive reactions to most lectins used. The Golgi region of the luminal secretory epithelial cells expresses a complex glycoconjugate pattern, as shown by its strong reactions to Man-(PSA, GNA), beta-GlcNAc-(S-WGA, PWA, DSA, UDA), beta-Gal-(RCA-I and -II), alpha/beta-GalNAc-(SBA, Jac, VVA, BPA) and complex NeuAc-(SNA) specific lectins, indicating that the secretory epithelial cells are active in glycosylation and secretion process. It was also shown in the present study that the basal and luminal epithelial cells are different in their glycoconjugates. The basal epithelial cells are rich in NeuAc(alpha2,3)Gal residues as they are stained specifically by MAA. The fibroblasts in the epithelial-smooth muscle interface and the smooth muscle cells close to the glandular epithelium are shown to express more glycoconjugates as they are stained intensely by GS-I-B4, GS-II and SBA. However, their role in the epithelial-stromal interaction in the seminal vesicle remains to be elucidated. In summary, the present study reports for the first time on the lectin binding patterns of the guinea pig seminal vesicle, and the results show that the seminal vesicle epithelium elaborates and secretes glycoconjugates in a complex pattern. Some of the lectins might be useful as histochemical markers for the secretory activity and specific structural components in the guinea pig seminal vesicle.

Acetylgalactosamine↗

Cytoskeletal-membrane interactions: a stable interaction between cell surface glycoconjugates and doublet microtubules of the photoreceptor connecting cilium.

The ciliary base is marked by a transition zone in which Y-shaped cross-linkers extend from doublet microtubules to the plasma membrane. Our goal was to investigate the hypothesis that the cross-linkers form a stable interaction between membrane or cell surface components and the underlying microtubule cytoskeleton. We have combined Triton X-100 extraction with lectin cytochemistry in the photoreceptor sensory cilium to investigate the relationship between cell surface glycoconjugates and the underlying cytoskeleton, and to identify the cell surface components involved. Wheat germ agglutinin (WGA) binds heavily to the cell surface in the region of the Y-shaped cross-linkers of the neonatal rat photoreceptor cilium. WGA binding is not removed by prior digestion with neuraminidase and succinyl-WGA also binds the proximal cilium, suggesting a predominance of N-acetylglucosamine containing glycoconjugates. Extraction of the photoreceptor plasma membrane with Triton X-100 removes the lipid bilayer, leaving the Y-shaped crosslinkers associated with the axoneme. WGA-binding sites are found at the distal ends of the crosslinkers after Triton X-100 extraction, indicating that the microtubule-membrane cross-linkers retain both a transmembrane and a cell surface component after removal of the lipid bilayer. To identify glycoconjugate components of the cross-linkers we used a subcellular fraction enriched in axonemes from adult bovine retinas. Isolated, detergent-extracted bovine axonemes show WGA binding at the distal ends of the cross-linkers similar to that seen in the neonatal rat. Proteins of the axoneme fraction were separated by SDS-PAGE and electrophoretically transferred to nitrocellulose. WGA labeling of the nitrocellulose transblots reveals three glycoconjugates, all of molecular mass greater than 400 kD. The major WGA-binding glycoconjugate has an apparent molecular mass of approximately 600 kD and is insensitive to prior digestion with neuraminidase. This glycoconjugate may correspond to the dominant WGA-binding component seen in cytochemical experiments.

Animals↗

Expression of glycoconjugates in human eustachian tubes with otitis media.

OBJECTIVES: To characterize glycoconjugate expression in normal human eustachian tubes and study the alterations in glycoconjugate expression found in eustachian tubes with otitis media. STUDY DESIGN: Using lectin histochemistry, alterations in glycoconjugates were studied in three normal temporal bones, in four temporal bones with mucoid otitis media (MOM), and in five with serous otitis media (SOM). METHODS: Sections of previously processed temporal bones were decelloidinized, and then incubated with seven biotinylated lectins--WGA, SNA, MAA, BPA, PNA, UEA-1, and LcH--that reflect seven carbohydrate residues of glycoconjugates, respectively: GlcNAc/NeuNAc, NeuNAc alpha(2-6)GalNAc, NeuNAc alpha(2-3)GalNAc, Gal beta(1-3) GalNAc, L-fucose, and alpha-mannose residues. Control sections were incubated with inhibitory carbohydrates or without biotinylated lectins. RESULTS: In the normal temporal bones, five carbohydrate residues in goblet cells and cilia of the eustachian tube demonstrated moderate to strong activity--NeuNAc alpha(2-6)GalNAc, NeuNAc alpha(2-3)GalNAc, GalNAc, Gal beta(1-3)GalNAc, and L-fucose. Two residues demonstrated weak activity--GlcNAc/NeuNAc and alpha-mannose. Temporal bones with MOM revealed increases in sialic acid and alpha-mannose, and a decrease in L-fucose. Residues of carbohydrates in the cilia of bones with SOM were notably decreased, especially for GalNAc, Gal beta(1-3)GalNAc, and NeuNAc alpha(2-6)GalNAc. CONCLUSIONS: Glycoconjugates in the normal human eustachian tube are rich in GalNAc, Gal beta(1-3)GalNAc, L-fucose, and NeuNAc alpha(2-3/2-6) GalNAc, but low in alpha-mannose and sialic acid. Eustachian tubes from cases with SOM or MOM demonstrated alterations in glycoconjugate expression in cilia and goblet cells, which may reflect disorder of the carbohydrate metabolism during otitis media, especially in SOM.

Eustachian Tube↗

Vagal control of mucus glycoconjugate secretion into the feline trachea.

1. We examined the effects of frequencies and patterns of electrical stimulation of the peripheral cut ends of the vagus nerves on the release of mucus glycoconjugates into feline trachea in vivo. Mucus glycoconjugates, radiolabelled biosynthetically with [35S]sulphate and [3H]glucose, were washed from a tracheal segment in situ, and dialysed before being counted and assayed chemically by the periodic acid-Schiff (PAS) method. 2. Vagal stimulation with regular pulses (10 V, 2 ms duration) at 1, 2.25, 4.5, 9 and 18 Hz produced frequency-dependent increases in the output of mucus glycoconjugates. 3. The muscarinic agonist pilocarpine (0.1-10 microM), given intrasegmentally, produced dose-dependent increases in the output of mucus glycoconjugates. 4. Pretreatment with atropine, phentolamine and propranolol reduced but did not abolish the effects of vagal stimulation. Vagus nerve stimulation still caused frequency-dependent increases in the output of mucus glycoconjugates. 5. High frequency stimulations at 22.5 and 47.5 Hz given intermittently (1 s burst then 4 s rest), whether in the absence or presence of cholinergic and adrenergic blockade, produced similar secretory responses as the same number of pulses delivered in regular trains at 4.5 and 9.5 Hz. This suggests that neither cholinergic nor non-adrenergic, non-cholinergic (NANC) nerve mechanisms in this system are potentiated by high frequency, intermittent burst stimulation. 6. In the absence of atropine, regular vagal stimulation had a greater effect on heart rate than did the same number of pulses delivered in bursts. 7. High molecular weight glycoconjugates from secretions were taken from the void volume of a Sepharose CL-2B gel filtration column and separated further by density-gradient centrifugation. Macromolecular components were observed at two densities, a typical mucin at 1.52 g ml-1, and a high density atypical component at 1.63 g ml-1. In secretions collected during vagal stimulation, either in the absence or presence of cholinergic and adrenergic blockade, the ratio of low density to high density macromolecules was higher than in unstimulated secretions. This can be explained if both cholinergic and NANC nervous vagal mechanisms stimulate the output of typical (density = 1.52 g ml-1) mucins into the feline trachea.

Adrenergic Fibers↗

Transient cholinergic glycoconjugate secretion from swine tracheal submucosal gland cells.

By inference, muscarinic stimulation of glycoconjugate release from tracheal submucosal gland cells appears to be a transient, nonequilibrium process (J. M. Farley and T. M. Dwyer, Life Sci. 48: 59-67, 1991). To directly characterize the release kinetics of glycoconjugate, we developed an enzyme-linked lectin assay (ELLA) of much improved precision and resolution. To collect secreted products with an improved time resolution, freshly isolated swine tracheal submucosal gland cells were continuously superfused with medium 199 at 37 degrees, buffered with N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) and CO2 or bicarbonate; fractions were collected every 15 s to 2 min. A 30-s pulse of 30 nM acetylcholine (ACh) increased the rate of glycoconjugate release by 10- to 25-fold for 2-3 min. The peak response averaged 14.2 +/- 9.0 ng protein.ml-1.min-1. 4 million cells-1 or 3.6 +/- 2.3 fg.cell-1.min-1 for 30 nM ACh and 16.2 +/- 3.0 ng protein.ml-1.min-1 for 100 nM ACh. There was no significant glycoconjugate release following a 30-s pulse of either 10 nM or 1 microM ACh. A second pulse after 7 min had no measurable effect on glycoconjugate release but a full response was obtained after 30 min. A continuous superfusion begun 1 min following the 30-s pulse resulted in a greater release of glycoconjugate than the pulse alone, but the response was not sustained, falling to twice basal levels within 5 min. We conclude that a brief muscarinic stimulation causes a triggered release of mucus glycoprotein followed by a relative refractory period.

Acetylcholine↗

Histochemistry of glycoconjugates in the goat nasolabial skin with special reference to eccrine glands.

The histochemistry of glycoconjugates in the nasolabial skin of the goat has been studied by means of a series of selected methods of light microscopy. The epidermis of the nasolabial skin was found to contain neutral and acid glycoconjugates with different saccharide residues. The secretory epithelial cells and secretory substances of the sebaceous glands contained primarily neutral glycoconjugates, whereas those of the apocrine glands involved largely strongly acidic and neutral glycoconjugates. In the epithelial cells and secretory substances of the nasolabial eccrine glands, glycoconjugates involved were characteristically strongly neutral but weakly acidic in nature. From the present results, the histophysiological significance of glycoconjugates in the particular histologic structures of the nasolabial skin has been discussed with special reference to the functions of this particular skin type in the goat.

Animals↗

Direct inhibitory action of glucocorticoid on glycoconjugate secretion from airway submucosal glands.

The precise mechanism by which glucocorticoids inhibit airway mucus secretion is still unknown. To study directly the effect of glucocorticoid on submucosal gland secretion, we examined the effects of dexamethasone on the precursor uptake, biosynthesis, and release of mucus glycoprotein in isolated feline tracheal submucosal glands. Mucus glycoprotein release from isolated glands was estimated by measuring [3H]glucosamine-labeled trichloroacetic acid (TCA)-precipitable glycoconjugates secreted into the medium. Released glycoconjugate per hour per dry weight of gland tissue was less than 7% of the total intracellular content, where intracellular content is defined as total 3H activity in the dissolved gland tissue. Treatment with 10(-9) to 10(-5) M dexamethasone for 24 to 72 h significantly reduced basal glycoconjugate secretion up to 22% of control (a 78% decrease) in a dose-dependent fashion, whereas the total intracellular 3H content was reduced to 70% of control (a 30% decrease) with no statistically significant differences from controls. The ratio of released glycoconjugates to the total intracellular content decreased significantly to 31% of control (a 69% decrease) after the treatment with 10(-10) to 10(-5) M dexamethasone. Further, ratio of radioactivity of TCA-precipitable glycoconjugates in the dissolved gland tissue to the total intracellular 3H content increased from 40% in nontreated controls to 46% after the treatment with dexamethasone (10(-5) M). Dexamethasone also inhibited the glycoconjugate secretion stimulated by dibutyryl cyclic AMP and alpha- and beta-adrenergic agonists. Simultaneously, the ratio of released to total intracellular content also decreased significantly after dexamethasone treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Recent developments in the synthesis and discovery of oligosaccharides and glycoconjugates for the treatment of disease.

The development of the first automated oligosaccharide synthesizer, along with new methods for screening carbohydrate ligand arrays is likely to lead to a rapid acceleration in both our ability to synthesize these molecules, and understand the roles of oligosaccharides and glycoconjugates in biology. Consequently we may uncover new avenues for therapeutic intervention more rapidly. These recent developments are very important since our understanding of the role of glycoconjugates in nature has traditionally fallen far behind that of the other biopolymers such as proteins and nucleic acids as the formation of, for example, glycosylated proteins is not template driven. The chemical synthesis of oligosaccharides and glycoconjugates has provided us with new potential cancer vaccines, antibiotics and new biotechnological tools. Glycobiologists have employed many such tools to uncover new signalling roles for oligosaccharides and glycoconjugates. In this review we aim to highlight some emerging methods for glycoconjugate assembly and screening, and discuss innovative approaches to glycoconjugate based drug design and delivery, all of which are, and will continue to be, fruitful avenues for medicinal chemistry research.

Animals↗

A glycoconjugate from corms of saffron plant (Crocus sativus L.) inhibits root growth and affects in vitro cell viability.

A glycoconjugate has been characterized from saffron corms (Crocus sativus L.) that inhibits the growth of roots of Nicotiana tabacum and Arabidopsis thaliana, at concentrations ranging from 1-100 micrograms m-3. Roots of seedlings grown in the presence 0.1 microgram m-3 glycoconjugate showed bulging of epidermal cells, whereas at 10 micrograms m-3, roots were completely devoid of hairs. At 100 micrograms m-3 glycoconjugate the cell walls of the root vascular tissues were thicker and, overall, the vascular tissue was enlarged. In addition, this glycan is cytotoxic to isolated tobacco cells and protoplasts, with 50% cell death induced by 0.5 and 2 micrograms m-3 glycoconjugate, respectively. Morphological and biochemical changes induced by the exposure to the glycoconjugate included cell size decrease, loss of regular cell shape, cytoplasm collapse, and release of intracellular proteins. This molecule at low concentrations (0.1 microgram m-3) mimics the effects of Yariv phenylglycosides and of mutant Arabidopsis which present defective or missing arabinogalactan-proteins (AGPs) in roots, indicating the glycoconjugate might interact with cell surface AGPs.

Arabidopsis↗

Ultrastructural cytochemistry of glycoconjugates in basophils from humans and animals.

The distribution of glycoconjugates in the basophil granules of humans, guinea pigs, and rabbits was compared. The observation of acid mucopolysaccharides using the dialyzed iron method and of sulfated glycoconjugates using the high iron diamine method revealed three types of reactions in the basophil granules of all three species: granules showing a strong overall reaction, granules showing reaction only at their periphery, and granules showing no reaction. With regard to the relationship between maturation and the types of basophil granules, it appeared that, in general, there were many type-1 granules among immature basophils, but that these granules decreased in mature basophils as type-3 granules increased. The reaction patterns of periodate-reactive neutral glycoconjugates, as shown by the periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) method, were different from those of acid mucopolysaccharides: the reaction of basophil granules was diffusely positive, and localization at the periphery was rarely observed. Therefore, unlike the acid mucopolysaccharides, it was difficult to classify the glycoconjugates into three types. However, as with acid mucopolysaccharides, there was a tendency for periodate-reactive glycoconjugates to decrease as maturation progressed. In terms of different species of animals, the reaction of periodate-reactive glycoconjugates with PA-TCH-SP was stronger in humans and rabbits than in guinea pigs.

Animals↗

Identifying glycoconjugate-binding domains. Building on the past.

The molecular details of how glycoconjugate-binding proteins interact with their ligands have been revealed by a variety of techniques. For example, proteases, chemical-modifying reagents and antibodies have served as effective probes of lectin functional domains. Protein crystallography has providing insight into how lectins are structured, and aided in determining which amino acids in these proteins are positioned appropriately for bond formation with glycoconjugates. In addition, the characterization and sequencing of naturally occurring, non-functional lectin variants have led to the identification of amino acids which play critical roles in a lectin's glycoconjugate-binding domain. Similarly, studies of lectin mutants produced by site-directed mutagenesis, and of synthetic peptides that mimic lectin binding properties, have demonstrated the importance of particular amino acids for glycoconjugate binding. An alternate approach to understanding lectin functional domains has been to compare the primary sequences of these proteins to reveal common sequence elements which allow them to be organized into families. For example, the discovery of amino acid homologies dispersed over long segments of the primary sequences of several lectins has suggested that many of these proteins have a related three-dimensional organization. In addition, the identification of more highly focused regions of sequence homology has indicated that many structures within the lectin glycoconjugate-binding domains themselves may be conserved. Scanning protein data banks for sequences homologous to known lectins has led to the identification of several previously unrecognized lectins, and aided in determining what portions of these proteins function in their glycoconjugate-binding domains.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

Analysis of glycoconjugate patterns of normal and hormone-induced dysplastic Noble rat prostates, and an androgen-independent Noble rat prostate tumor, by lectin histochemistry and protein blotting.

BACKGROUND: Alteration of the expression of glycoconjugates is frequently observed in tumors. However, studies on the changes of cellular glycosylation in the early premalignant stage of prostate carcinogenesis are scarce. METHODS: The present study characterized and compared the glycoconjugates expressed in the dysplastic lateral prostate induced in Noble (Nb) rat by steroid hormones and a transplantable androgen-independent Nb rat prostatic carcinoma line (AIT) by lectin histochemistry and protein blotting. RESULTS: The results of lectin histochemistry show that the dysplastic prostatic epithelium elaborates altered patterns of glycosylation, which are distinct from the normal secretory epithelium. Some individual cells in the dysplastic epithelium were intensely labeled by the N-acetylgalactosamine (GalNAc)-specific (agglutins from Glycine max [SBA], Helix aspera [HAA], Helix pomatia [HPA], Vicia villosa [VVA], Erythrina cristigalli [ECA]) and complex-type oligosaccharide-specific (Phaseolus vulgaris agglutin [PHA-E]) lectins, indicating that these cells contained abundant GalNAc(alpha1,3)GalNAc/Gal and Gal(beta1,4)GlcNAc(alpha1,2)Man(alpha1,6) residues. These lectins also bound to some tumor cells in the AIT, suggesting that these sugar residues are common in some dysplastic and neoplastic prostatic cells. The study has also identified several lectins (agglutins from Griffonia simplicifolia [GS-I-B4], Arachis hypogaea [PNA], Ricinus communis [RCA-I], Maackia amurensis [MAA], Sambucus nigra [SNA]), which bound only to some AIT tumor cells but not to dysplastic epithelium, indicating that alpha/betaGal and sialic acid-containing glycoconjugates are expressed by neoplastic prostatic cells. The results of lectin blottings with Triticum vulgare agglutin [S-WGA] Ulex europaeus agglutin [UEA-I] and PHA-E have identified five major glycoprotein bands (of apparent molecular weights of 116, 79, 64, 61, and 57 kDa) in the microsomal fraction of testosterone plus 17beta-estradiol (T + E2)-treated lateral prostate. These lectin-reactive bands were not detected in the AIT extracts. In the AIT microsomal extract, two glycoprotein bands of molecular weights of 58 and 46 kDa were revealed by SBA and PNA. CONCLUSIONS: The present study shows that there is an increased expression of GalNAc(alpha1,3)GalNAc/Gal residues and triantennary complex-type oligosaccharides in the dysplastic epithelial cells as compared to normal secretory epithelial cells in rat lateral prostate. This altered expression of glycoconjugates revealed in the dysplastic epithelium indicates an aberrant glycosylation in the early premalignant stage of prostate carcinogenesis. The results also show that the AIT tumor cells are heterogeneous in their glycoconjugates and different from the dysplastic epithelial cells.

Animals↗