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A comparative study of vitreous-body and zonular glycoconjugates that bind to the lectin from Ulex europaeus.

Eyes from adult rodents, rabbits and humans were fixed in formalin, embedded in paraffin and incubated with a horseradish peroxidase-conjugated lectin from Ulex europaeus to localize vitreous-body and zonular glycoconjugates. Rodent eyes had reaction product for peroxidase activity in fibrous structures in the posterior chamber, vitreous base and vitreous cortex. The zonules and the internal limiting membrane region of the retina also were stained. Rabbit eyes had more stained fibrous material in the vitreous base than rodent eyes and the attachment region of the zonules on the lens capsule, the anterior hyaloid membrane and tracts in the vitreous cortex were more heavily stained in rabbits. There was heavy staining of the thick vitreous base in the human eyes as well as staining of zonules, anterior hyaloid membrane and vitreous cortex. The localization of this lectin may be specific for fucose in glycoproteins or other glycoconjugates, although this was not demonstrated here. However, the location of lectin binding sites correlates well with known sites of uptake of tritiated fucose and tritiated glucosamine in rabbit eyes. Eyes from the larger species studied had more lectin-binding glycoconjugates in fibrous structures in the vitreous body than those from smaller species. The amount of glycoconjugate identified in some of the lectin-binding sites may be related to some extent to the degree of stress incident upon those sites.

Adult↗

The histochemistry of glycoconjugates in the colonic epithelium of the chicken.

In the colonic epithelium of the chicken, glycoconjugates have been studied by means of selected histochemical methods of light and electron microscopy. According to the results obtained, most of the colonic goblet cells contained acidic and neutral glycoconjugates with sulphate and vicinal diol groupings, alpha-D-mannose and alpha-D-glucose residues and sialic acid-galactose dimers. These goblet cells were found to undergo changes in histochemical reactivity during upward migration along the crypts; alpha-D-mannose and alpha-D-glucose residues and terminal sialic acid-galactose dimers increased in amount. The striated border of the colonic columnar cells has, likewise, been found to contain such glycoconjugates as were similar in reactivity to those of the goblet cells. The histophysiological significances of glycoconjugates involved in the chicken colonic epithelium have been discussed with special reference to the functional activities of the carbohydrates.

Animals↗

Sulfated glycoconjugates in rat incisor secretory ameloblasts and developing enamel matrix.

We investigated the ultrastructural distribution and histochemical properties of sulfated glycoconjugates, which could be preserved by glutaraldehyde fixation, in secretory ameloblasts and developing enamel matrix, by use of the high iron diamine thiocarbohydrazide silver proteinate (HID-TCH-SP) staining and enzymatic digestion methods. Large type HID-TCH-SP stain deposits, approximately 10 nm in diameter, were detected on the interdigitating cell membrane of Tomes' process, inside some secretory granules, on the lateral cell membrane of stratum intermedium, in the basement membranes associated with outer enamel epithelium and endothelial cells of capillary, within the so-called hole region, and in the enamel matrix near future enamel-cement junction. A few large type stain deposits were, however, randomly distributed over the whole layer of enamel matrix. Small type stain deposits smaller than 5 nm in diameter were localized within some secretory granules and Golgi vesicles of ameloblasts and on the surface layer of developing enamel matrix. While the large type HID-TCH-SP stain deposits associated with the basement membranes and on the lateral cell membrane of stratum intermedium were susceptible to heparitinase, the others resisted enzymatic digestion not only by heparitinase but also by testicular hyaluronidase and chondroitinase ABC, indicating that they represent sulfated glycoconjugates other than heparan sulfate, chondroitin sulfate A, dermatan sulfate, or chondroitin sulfate C. On the other hand, HID-TCH-SP stain deposits within the secretory granules of odontoblasts and in the predentine matrix were susceptible to testicular hyaluronidase. Thus, it was confirmed that the composition of sulfated glycoconjugates secreted into the developing enamel matrix differs essentially from that of sulfated glycoconjugates associated with dentinogenesis.

Ameloblasts↗

Developmentally regulated plasmalemmal glycoconjugates of the surface and neural ectoderm.

The plasmalemmal glycoconjugates of the ectoderm surrounding the rat embryo's caudal neuropore were mapped at the ultrastructural level, using various lectin probes. These included the agglutinins of wheat germ, soybean, Ricinus communis, Lotus tetragonolobus, and Canavalia ensiformis. Each lectin produced a characteristic binding pattern. Comparison of precursor cells of surface ectoderm, neural crest, and neural tube revealed that, even prior to neural tube formation, these three cell types can be distinguished by the sets of lectin receptors they express on their apical plasmalemma. The high density of lectin receptors found at the open neural groove level decreases dramatically during neurulation. Further changes in surface glycoconjugates must occur during neuronal differentiation because sprouting neurons exhibit yet another lectin binding pattern (K.H. Pfenninger, M.-F. Maylié-Pfenninger, L. B. Friedman, and P. Simkowitz, 1984, Dev. Biol. 106, 97-108). These results indicate that the commitment of ectodermal cells to diverging lineages (epidermis, neural crest, and tube) is reflected in their surface carbohydrates and occurs while they are still part of a continuous epithelial sheet. Furthermore, the plasmalemmal glycoconjugates of the ectoderm are developmentally regulated, and particularly dramatic changes in glycoconjugates expression are linked to neurulation.

Animals↗

Glycoconjugate synthesis during early pregnancy: hyaluronate synthesis and function.

The synthesis of various glycoconjugate classes by mouse uteri during the pre- and peri-implantation period has been examined using [3H]glucosamine as a metabolic precursor. A unique and dramatic (five- to sixfold) increase was observed in the synthesis of hyaluronate on the day upon which embryo implantation normally occurs. Mated, but nonpregnant mice did not display increased hyaluronate biosynthesis. In contrast to hyaluronate, the synthesis of most other types of glycoconjugates remained fairly constant during the first 5 days of pregnancy. Low (1500-5000)-molecular-weight N-linked oligosaccharides constituted the major class of oligosaccharides synthesized under all conditions. High (greater than 10,000)-molecular-weight glycoconjugates constituted the second most abundant class of glycoconjugates synthesized (20-30%). Most (85%) of the newly synthesized hyaluronate was associated with the nonepithelial cell types of the uterus. Experiments using ovariectomized mice receiving steroid hormones demonstrated that uterine hyaluronate synthesis was induced preferentially by an artificial decidual stimulus and implicated stromal cells as the site of hyaluronate synthesis. In addition, it was demonstrated that tissue culture plates coated with hyaluronate, but not other polysaccharides, support attachment and spreading of a large fraction (60 to 70%) of embryos cultured in serum-free medium. Collectively, these studies indicate that increased hyaluronate biosynthesis accompanies decidual responses in the endometrium and may promote embryo implantation following initial penetration of the uterine epithelium.

Animals↗

Characterisation of sugar residues in glycoconjugates of pig mandibular gland by traditional and lectin histochemistry.

Sugar residues are important components of salivary gland secretion. Traditional histochemical methods and lectin histochemistry were used to characterise glycoconjugates present in the mandibular gland of normal adult pigs. Acinar cells contained abundant quantities of glycoconjugates with the terminal trisaccharide sialic acid - (alpha 2-->3, 6) galactosyl (beta 1-->3) N -acetylgalactosamine. Mandibular acinar cells also contained alpha and beta N -acetylgalactosamine and N -acetylglucosamine residues, whereas the demilunar cells contained glycoconjugates with fucose, mannose and N -acetylglucosamine residues. In the duct system a range of sugar residues were localised throughout the cell cytoplasm or limited to the apical surface. These results provide new knowledge concerning the structure of salivary glycoconjugates in normal adult pig and a basis for future pathological studies.

Acetylglucosamine↗

Selective expression of endogenous lactose-binding lectins and lactoseries glycoconjugates in subsets of rat sensory neurons.

Cell-surface glycoconjugates and endogenous lectins have been implicated in cellular interactions that contribute to embryonic development. Functional subsets of primary sensory neurons in mammalian dorsal root ganglia (DRG) have been shown recently to express specific cell-surface oligosaccharide structures. We report here that endogenous lectins with affinity for sensory neuron glycoconjugates are also synthesized by subsets of DRG neurons and are present in the dorsal horn of the developing spinal cord. The distribution of two endogenous lactose-binding lectins, RL-14.5 and RL-29 (subunit Mrs of 14,500 and 29,000, respectively), was examined by immunoblotting and by immunocytochemistry in embryonic and postnatal rat DRG and spinal cord. The two lectins appear soon after the formation of the DRG and are present in the cell bodies and terminals of subsets of DRG neurons that also express cytoplasmic and cell-surface lactoseries glycoconjugates. RL-14.5 and RL-29 are present in overlapping, but not coincident, subsets of DRG neurons that project to the superficial dorsal horn of the spinal cord. In addition, RL-14.5, but not RL-29, is expressed in spinal motoneurons from embryonic day 14. The preferential localization of lactoseries glycoconjugates and lactose-binding lectins in the DRG and the dorsal horn of the spinal cord suggests that these complementary molecules contribute to the development and function of primary sensory neurons.

Animals↗

Enzymatic large-scale synthesis of MUC6-Tn glycoconjugates for antitumor vaccination.

In cancer, mucins are aberrantly O-glycosylated, and consequently, they express tumor-associated antigens such as the Tn determinant (alpha-GalNAc-O-Ser/Thr). As compared with normal tissues, they also exhibit a different pattern of expression. In particular, MUC6, which is normally expressed only in gastric tissues, has been detected in intestinal, pulmonary, colorectal, and breast carcinomas. Recently, we have shown that the MCF7 breast cancer cell line expresses MUC6-Tn glycoproteins in vivo. Cancer-associated mucins show antigenic differences from normal mucins, and as such, they may be used as potential targets for immunotherapy. To develop anticancer vaccines based on the Tn antigen, we prepared several MUC6-Tn glycoconjugates. To this end, we performed the GalNAc enzymatic transfer to two recombinant MUC6 proteins expressed in Escherichia coli, using UDP-N-acetylgalactosamine: polypeptide N-acetylgalactosaminyltransferases (ppGalNAc-Ts), which catalyze in vivo the Tn antigen synthesis. We used either a mixture of ppGalNAc-Ts from MCF7 breast cancer cell extracts or a recombinant ppGalNAc-T1. In both cases, we achieved the synthesis of MUC6-Tn glycoconjugates at a semi-preparative scale (mg amounts). These glycoproteins displayed a high level of Tn antigens, although the overall density depends on both enzyme source and protein acceptor. These MUC6-Tn glycoconjugates were recognized by two anti-Tn monoclonal antibodies that are specific to human cancer cells. Moreover, the MUC6-Tn glycoconjugate glycosylated using MCF7 extracts as the ppGalNAc-T source was able to induce immunoglobulin G (IgG) antibodies that recognized a human tumor cell line. In conclusion, the large-scaled production of MUC6 with tumor-relevant glycoforms holds considerable promise for developing effective anticancer vaccines, and further studies of their immunological properties are warranted.

Antigens, Neoplasm↗

Glycoconjugates in sweat glands and other structures of skin from normal and cystic fibrosis subjects.

In this study, glycoconjugates of human skin varied in structure between cell types in an individual and often between individuals for a given cell type. Stored secretory material in dark cells of the sweat gland coil contained complex carbohydrate with terminal alpha-galactose, N-acetylgalactosamine, fucose, or sialic acid. The plasmalemma of clear cells in the secretory coil stained conspicuously for terminal alpha-N-acetylgalactosamine, and the cytosol of clear cells contained lectin-reactive glycogen in the majority of specimens. Superficial and deep cells of the sweat duct evidenced plasmalemmal glycoconjugate with terminal N-acetylgalactosamine. However, only the superficial cell plasmalemma in the duct stained for terminal alpha-N-acetylgalactosamine, fucose, and sialic acid. In several specimens, only the deep cells in the sweat duct revealed plasmalemmal glycoconjugate with terminal beta-galactose. Sebaceous glands alone displayed lectin affinity demonstrative of terminal alpha-galactose and like other sites stained for terminal beta-galactose and alpha-N-acetylgalactosamine. The epithelium in epidermis and hair follicles appeared similar, except for epidermis failing to evidence fucose and alpha-N-acetylgalactosamine with certain lectins. Both underwent changes in glycoconjugate composition with cell maturation. Skin from control subjects and patients with cystic fibrosis did not differ in lectin-binding properties. Abnormalities observed in cystic fibrosis specimens included decreased volume of sebaceous glands and, in two cases, increased infiltration of macrophages staining for terminal N-acetylgalactosamine.

Acetylgalactosamine↗

Cyclic nucleotides fail to affect mucus glycoconjugate secretion from canine tracheal explants.

Canine tracheal explants, incubated overnight with [3H]glucosamine, elicited an enhanced secretion of ethanol-precipitated [3H]labelled glycoconjugate when challenged with methacholine, 10 microM. Neither the beta-adrenoceptor agonist isoprenaline, 10 microM, nor the phosphodiesterase inhibitor theophylline, 10 mM, had any significant effect on glycoconjugate secretion. Dibutyryl cyclic AMP, 1 mM, and dibutyryl cyclic GMP, 1 mM, alone or in combination with theophylline, 10 mM, were devoid of activity on unstimulated or methacholine-stimulated tracheal explants. The calcium ionophore A23187, 10 microM, stimulated [3H]glycoconjugate secretion from each of the tissues tested; however, the cyclic nucleotides failed to modify this response. These data indicate that the cyclic nucleotides play little, if any, role in mucus glycoconjugate secretion by the canine trachea.

Animals↗

Cognate stimulatory B-cell-T-cell interactions are critical for T-cell help recruited by glycoconjugate vaccines.

Covalent linkage of a bacterial polysaccharide to an immunogenic protein greatly enhances the carbohydrate's immunogenicity and induces polysaccharide-specific B-cell memory in vivo. These findings have spurred the development of glycoconjugate vaccines for serious bacterial infections. The specific B-cell-T-cell interactions responsible for recruitment of T-cell help by glycoconjugate vaccines are not well defined. We used mice deficient in molecules critical for stimulatory, cognate B-cell-T-cell interactions to study how T cells improve the immunogenicity of a glycoconjugate vaccine against group B streptococcal disease. Isotype switching to immunoglobulin G (IgG) was abrogated in mice deficient in major histocompatibility complex (MHC) class II antigen (Ag)-T-cell receptor (TCR), B7-CD28, or CD40-CD40L interactions. However, expression of either the B7-1 or the B7-2 molecule on antigen-presenting cells was sufficient for optimal T-cell costimulation. T cells activated by the vaccine also played a pivotal role in determining the magnitude of the IgM response to the polysaccharide. Comparable results were obtained with pathway antagonists. These data suggest that MHC class II Ag-TCR, B7-CD28, and CD40-CD40L interactions are critical for immune responses to glycoconjugate vaccines in vivo.

Animals↗

Isolation and partial characterization of a high molecular weight anionic glycoconjugate from transforming growth factor-beta treated bovine articular chondrocyte cultures.

A high molecular weight anionic glycoconjugate was isolated from the media of the transforming growth factor-beta treated chondrocyte cultures by anion-exchange chromatography on DEAE-Sephacel and Mono Q columns and was partially characterized. This high molecular weight anionic glycoconjugate was not detected in the non-treated (control) cultures. Characterization studies showed that the glycoconjugate is a non-reducible, non-collagenous glycoprotein containing O-linked, N-linked, and sialic acid substituted carbohydrate units. The isolated glycoconjugate stained "blue" with Stains All and migrated as a single band on sodium dodecyl sulfate gradient gels (2.5-10% acrylamide - diallyl tartardiamide) at an estimated molecular weight of 540 000. Amino acid and amino sugar analyses showed that it is rich in aspartic acid--asparagine, glutamic acid--glutamine, alanine, proline, and glycine, and contains galactosamine and glucoasmine. This transforming growth factor-beta inducible glycoprotein may be involved in cell differentiation and in the cartilage repair process. It may also be used as a marker to localize the biological activity of transforming growth factor-beta in articular cartilage.

Amino Acids↗

NANC nerve pathways controlling mucus glycoconjugate secretion into feline trachea.

We used autonomic-blocking drugs to define nonadrenergic noncholinergic (NANC) vagus nerve pathways regulating tracheal mucus secretion. In anesthetized cats, mucus glycoconjugates, radiolabeled biosynthetically with [35S]sulfate and [3H]glucose, were washed from a tracheal segment in situ and dialyzed before scintillation counting and chemical assay with periodic acid-Schiff (PAS). Without autonomic blockade, vagal stimulation (9.5 Hz, 10 V, 2-ms pulse width, 10-min duration) increased outputs of radiolabeled and PAS-reactive glycoconjugates repeatably over four stimulation periods. In other animals, vagus nerves were stimulated with administration of autonomic blockers between stimulations. The first stimulation (no blockers) increased glycoconjugate output (delta 35S = 221 +/- 43.3%, delta 3H = 58 +/- 13.8%; delta PAS = +299 +/- 82.7%). Atropine, phentolamine, and propranolol reduced these responses (delta 35S = 67 +/- 15.6%; delta 3H = 26 +/- 5.3%; delta PAS = 88 +/- 25.6%). Guanethidine did not significantly lessen them further, although delta 3H was no longer significant. Ganglion blockade with hexamethonium prevented most of the remaining response to vagal stimulation (P less than 0.05 for diminution of delta 35S and delta PAS), but small effects persisted (delta 35S = 17 +/- 5.6%; delta PAS = 20 +/- 6.8%; P less than 0.05). We conclude that the main NANC vagal pathway controlling tracheal glycoconjugate secretion runs orthodromically.

Adrenergic Antagonists↗

Inhibition by vasoactive intestinal peptide of glycoconjugate and lysozyme secretion by human airways in vitro.

The effects of vasoactive intestinal peptide (VIP) were analyzed on the in vitro release of radioactively labeled mucus glycoconjugates and lysozyme by explants of human bronchial mucosa from normal subjects and from patients with chronic bronchitis. These effects were compared with the effects of VIP on the discharge of labeled macromolecules (analyzed by quantitative autoradiography) from mucous and serous cells of the airway submucosal glands. In explants of 9 mucosal specimens of normal airways, VIP (10 ng to 1 micrograms/ml) caused a dose-dependent inhibition of baseline and methacholine-stimulated release of both glycoconjugates and lysozyme. At a concentration (1 micrograms/ml) that caused maximal inhibition of glycoconjugate and lysozyme release, VIP also caused a small inhibition of baseline but not methacholine-induced discharge of labeled macromolecules from mucous and serous cells of the submucosal glands. In explants from 5 patients with chronic bronchitis, VIP did not inhibit baseline or methacholine-stimulated glycoconjugate release and mucous or serous cell discharge, even at doses greater than 1 micrograms/ml. By contrast, VIP did inhibit baseline and methacholine-stimulated release of lysozyme, but this was less marked than in explants of normal airways. In view of the proximity of neurons containing VIP to submucosal gland cells, this study supported the hypothesis that VIP may contribute to the neurohumoral regulation of mucus secretion by the human airway. It was evident, however, that the effects of VIP could not be accounted for in terms of inhibiting cell discharge alone. In chronic bronchitis, the reduction or absence of sensitivity to VIP inhibition suggests a functional difference in the regulation of mucus secretion, which may contribute to mucus hypersecretion.

Bronchi↗

Developmental changes in glycoconjugate secretion by ferret tracheas.

Surface epithelium and submucosal glands of the ferret trachea undergo extensive postnatal development. This study examined developmental changes in rates of release and types of high molecular weight glycoconjugates secreted by explanted ferret tracheas. Digestion with bovine testicular hyaluronidase separated the high molecular weight glycoconjugates into two types, hyaluronidase-resistant mucins and hyaluronidase-susceptible glycosaminoglycans. Release rates were measured under unstimulated conditions and in the presence of known secretagogues. The unstimulated rate of release of total 3H-glycoconjugates was 4-fold higher at birth than after complete maturation. The mucin content varied from 39 to 74% of total 3H-glycoconjugates; however, no age-related pattern was observed for mucin/glycosaminoglycan ratios. The rate of release of 3H-mucins was 6-fold higher at birth than in the adult but rapidly dropped to adult levels by 28 days of age. The secretory cells in the tracheal epithelium of newborn ferrets had more abundant rough endoplasmic reticulum than did mature goblet cells, suggesting increased synthesis of secretory product. Response to methacholine and trypsin, both known stimulators of mucin release, was not observed until 28 and 54 days of age, respectively. Incorporation of 35S-sulfate into mucins relative to that for 3H-glucosamine increased with age, consistent with increasingly acidic histochemical staining properties of secretory cells. These developmental differences in rates of release, modulation of release, and relative sulfation of mucins may represent changes in secretory and synthetic mechanisms of the secretory cells.

Age Factors↗

Histochemical localization of galactose-containing glycoconjugates in sensory neurons and their processes in the central and peripheral nervous system of the rat.

We studied the distribution of sugar residues in the oligosaccharide chains of complex carbohydrates in tissue sections of rat spinal cord, brainstem, and sensory ganglia using twelve lectin-horseradish peroxidase conjugates. Glycoconjugates containing terminal galactose residues were localized apparently in the Golgi apparatus in a population of predominantly small B-type neurons in spinal and trigeminal ganglia. Large A-type neurons rarely showed reactivity with galactose-binding lectins. A cells stained for glycoconjugates with N-glycosidically linked oligosaccharides and glycogen. The central and peripheral processes of the small neurons, mostly unmyelinated C fibers in sensory roots and spinal nerves, contained an abundance of glycoconjugates with terminal alpha-galactose residues. The central projections and terminals of small to medium-sized primary sensory neurons in the spinal and trigeminal ganglia were visualized in Lissauer's tract and the substantia gelatinosa in the spinal cord, and in the spinal trigeminal tract and the nucleus trigeminus in the lower medulla with lectins specific for terminal alpha-galactose residues. In addition, fibers of the solitary system and the area postrema were reactive with these lectins. The peripheral and central nervous system elements with affinity for galactopyranosyl-specific lectins correspond in distribution with neuroanatomical regions thought to be involved in the transmission and relay of somatic and visceral afferent inputs such as pain and temperature. Such specific localization of a glycosubstance to a distinct subpopulation of neurons and their peripheral and central processes suggests that the particular glycoconjugate may be of physiological significance.

Animals↗

Interspecific variation of zona pellucida glycoconjugates in several species of marsupial.

The zona pellucida glycoconjugate content of several marsupial species was investigated using differential lectin histochemistry. Ovaries from fat-tailed dunnarts, a southern brown bandicoot, grey short-tailed opossums, brushtail possums, ringtail possums, koalas and eastern grey kangaroos were fixed, embedded in paraffin wax, sectioned and stained with ten fluorescein isothiocyanate-conjugated lectins. Sections were also incubated with either neuraminidase or saponified, respectively, before incubation with the lectins to identify saccharide residues masked by sialic acids or O-acetyl groups on sialic acids. The zonae pellucidae surrounding the oocytes of the marsupials demonstrated interspecific variation in glycoconjugate content, with mannose-containing glycoconjugates exhibiting the greatest variation. Some of the zona pellucida glycoconjugates of all species, except those of the opossums, were masked by sialic acid with an increase in fluorescence with lectins from Arachis hypogea (PNA), and Glycine max (SBA), after desialylation. The disaccharide beta-galactose(1-4)N-acetyl-D-glucosamine appeared to be conformationally masked by O-acetyl groups of sialic acids in the zonae pellucidae of all species, with an increase in fluorescence with the lectin from Erythrina cristagalli (ECA), after saponification. Similar intensity and localization of beta-(1-4)-N-acetyl-D-glucosamine, as shown by staining of the lectin from Triticum vulgaris (WGA), to the inner and outer regions of the zona pellucida, were found to those reported in eutherian species. WGA fluorescence became uniform throughout the zonae pellucidae after saponification, indicating differential O-acetylation of sialic acids on the internal compartment of the zonae pellucidae.

Acetylation↗

Histochemical analysis of acidic glycoconjugates in the endothelium lining the splenic blood vessels in the rat.

An electron microscopic analysis with specific histochemical stainings for acidic glycoconjugates was carried out to examine the endothelium lining blood vessels of the rat spleen. Histochemical staining performed was the postembedding high or low iron diaminethiocarbohydrazide-silver protein-physical development (HID or LID-TCH-SP-PD) method, with or without prior digestion with acidic glycoconjugate-degrading enzymes, such as heparitinase, testicular hyaluronidase, chondroitinase B and neuraminidase. The results indicated that the acidic glycoconjugates in the basal lamina of the endothelial cells lining the four types of blood vessel (central arteries, arterial capillaries, splenic sinuses and pulp veins) were heparan sulfate, chondroitin sulfate A and/or C, chondroitin sulfate B and sialic acid residues. In the endothelial cells lining the central arteries, arterial capillaries and pulp veins, the surface coat of the luminal plasma membrane included heparan sulfate, chondroitin sulfate A and/or C, chondroitin sulfate B and sialic acid residues, whereas the corresponding ultrastructure of the splenic sinuses was devoid of detectable amounts of acidic glycoconjugates. This suggests that such characteristic histochemical features of the endothelium in the four types of the splenic blood vessel can be related to the possible physiological functions of the spleen.

Animals↗