The glycoprofile patterns of endothelial cells in usual interstitial pneumonia.

BACKGROUND: The pathological classification of cryptogenic fibrosing alveolitis has been a matter of debate and controversy for histopathologists. OBJECTIVE: To identify and specify the glycotypes of capillary endothelial cells in usual interstitial pneumonia (UIP) compared to those found in normal tissue. METHODS: Sections of formalin-fixed, paraffin-embedded blocks from 16 cases of UIP were studied by lectin histochemistry with a panel of 27 biotinylated lectins and an avidin-peroxidase revealing system. RESULTS: High expression of several classes of glycan was seen de novo in capillary endothelial cells from patients with UIP including small complex and bi/tri-antennary bisected complex N-linked sequences bolund by Concanavalin A and erythro-phytohemagglutinin, respectively, GalNAca1 residues bound by Helix pomatia and Maclura pomifera agglutinins, and L-fucosylated derivatives of type II glycan chains recognized by Ulex europaeus agglutinin-I. Glycans bound by agglutinins from Lycopersicon esculentum (ß1,4GlcNAc) and Wisteria floribunda (GalNAc) as well as GlcNAc oligomers bound by Phytolacca americana and succinylated Wheat Germ agglutinin were also seen in the capillary endothelial cells of UIP. In contrast, L-fucosylated derivatives of type I glycan chains were absent in cells from cases of UIP when Anguilla anguilla agglutinin was applied, unlike the situation in normal tissue. CONCLUSION: These results may indicate existence of two distinct populations of endothelial cell in UIP with markedly different patterns of glycosylation, reflecting a pattern of differentiation and angiogenesis, which is not detectable morphologically.

Glycophenotype of prostatic carcinomas.

The factors that affect the progression of prostatic carcinoma are poorly understood, but it is known that carbohydrate antigens on the tumour cell surface play a role in the transforming and metastatic processes. The present report aimed to perform a comparative, lectin-histochemical study of benign and carcinomatous prostates, using a battery of lectins, in combination with monoclonal antibodies against Lewis antigens, and a semi quantitative study, to investigate the changes in glycosylation patterns that occur in prostatic carcinoma. Blocks from 27 necropsy cases of prostatic carcinoma were sectioned and stained with H+E, fifteen biotinylated lectins chosen to probe for a wide range of oligosaccharide sequences within several categories of glycoprotein glycans, using a lectin-biotin avidin-peroxidase method, and monoclonal antibodies against Lewisa, sialyl Lewisa and sialyl Lewisx antigens. The glycophenotype of prostatic carcinoma differed from that of the noncancerous prostate in revealing more intense staining with the following lectins (AAA, UEA-1, DBA, WFA, VVA, HPA, BSA-1B4, MPA, ECA, AHA, and CTA), while the binding patterns of (GNA and NPA) were almost similar in both prostatic carcinoma and the noncancerous prostate. Lewis antigens are found to be expressed in prostatic carcinomas but not in the noncancerous prostate. The observations of this study suggest that the gylcophenotype of transformed prostatic cells was modified. It showed a moderate increase in, and changing patterns of, fucosylation and galactosylation, increased branching of side chains and sharp rise in 2 deoxy, 2 acetamido galactosylation and masking process by sialylation, especially by α2-3 and α2-6 linkages. All these changes in the glycosylation pattern of the transformed prostatic cells were observed on O-glycans, no changes were observed on N-glycans.

Lectin and other histochemical studies of the articular cartilage and the chondro-osseous junction of the normal human knee joint.

There are few studies on normal, adult diarthrodial joints which look in detail at the histochemical properties of the chondro-osseous junctional region. This study of the normal human knee joint was performed using lectin and other histochemical techniques. There were differences in the reactions of mineralised cartilage compared to those of hyaline cartilage with the former demonstrating more collagen and less glycosaminoglycans. Lectin histochemistry revealed more accessible terminal 2-deoxy,2-acetamido-alpha-D: -galactose and more N-acetyllactosamine but less fucosyl and alpha-2,6-linked-sialyl termini in the mineralised cartilage. The hyaline cartilage chondrocytes stained for N-glycans but those of mineralised cartilage did not. The staining patterns of prolongations and islands of uncalcified cartilage running through the calcified layer to abut bone and marrow spaces were distinct, resembling the patterns of the hyaline cartilage but with some unique features. A possible relationship was revealed between the presence of the Maclura pomifera ligand (Galbeta1,3GalNAcalpha1-) and mineralisation. Subchondral bone had a markedly restricted glycoprofile.

The tidemark of the chondro-osseous junction of the normal human knee joint.

The chondro-osseous junction includes the junction between calcified and non-calcified cartilage matrices often referred to as the tidemark. A detailed knowledge of the structure, function and pathophysiology of the chondro-osseous junction is essential for an understanding both of the normal elongation of bones and of the pathogenesis of osteoarthrosis. In this study the molecular anatomy of the tidemark was studied using histochemical techniques, including lectin histochemistry, on blocks of normal cartilage from human knee joints. The tidemark stained with H and E, picro-sirius red, toluidine blue, safranin O and methyl green, but not with alcian blue in the presence of magnesium chloride at 0.05 M or above. It stained with only four lectins, those from Datura stramonium, Maclura pomifera, Erythrina crystagalli and Helix pomatia, out of the 19 used. Therefore, it is rich in collagen and contains hyaluronan, but appears to lack the glycosaminoglycans of 'conventional' proteoglycans and it expresses a very limited and distinctive lectin staining glycoprofile, which is probably attributable to specific glycoproteins. In addition, the tidemark had a distinct microanatomical trilaminate appearance. From all of these results it is clear that this part of the chondro-osseous junctional region is chemically more complex and distinctive than has previously been described.

Application of solid-phase micro-extraction technology to drug screening and identification.

Benzodiazepines, tricyclic antidepressants and local anaesthetics are frequently involved in poisoning episodes and fatalities. A specific, sensitive and rapid procedure for identifying and quantifying such drugs in postmortem matrices has been developed using solid-phase micro-extraction (SPME) and gas chromatography-mass spectrometry. Very clean extracts were obtained in one step using SPME. The most commonly used fibre coatings were tested to select the best coating for SPME of the drugs. The appropriate fibre coating for most drugs was polyacrylate, followed by Carbowax-divinylbenzene. A Hewlett-Packard 5890 gas chromatograph in combination with a Trio 2000 mass spectrometer was used to analyse the samples. Temperature, time, pH and addition of sodium chLoride were optimized to obtain consistent extraction. The between-day and within-day coefficients of variation were less than 16% and less than 6%, respectively.

Glycosylation in the near-term epitheliochorial placenta of the horse, donkey and camel: a comparative study of interbreeding and non-interbreeding species.

Studies from this laboratory have shown great diversity in the glycosylation of tissues comprising the interhaemal barrier of species with different placental types. This diversity may be one of the factors preventing interbreeding between species. Glycan expression within the uterine epithelium and trophoblast of the interhaemal barrier was examined to test this proposition in three species with similar diffuse, microcotyledonary, epitheliochorial allantochorionic types of placenta: the horse (Equus caballus) and donkey (Equus asinus), which can interbreed with each other, and the camel (Camelus dromedarius), which cannot interbreed with either of the other two species. A panel of 14 lectins was used and it was found that glycosylation patterns were generally similar between placental tissues of the horse and donkey, except for the expression of non-bisected complex N-glycan and some sialic acids, whereas those of the camel showed striking differences in the binding of lectins to many structures carrying terminal residues of fucose, N-acetyl galactosamine and beta-galactose, as well as to complex N-glycans and sialic acids. These results are consistent with the proposition that interbreeding species carry similar glycans in tissues forming the interhaemal barrier whereas glycodiversity is one of the factors preventing implantation and subsequent placental development in interspecific hybrids.

Mycophenolic acid does not inhibit protein glycosylation in T lymphocytes.

BACKGROUND: Mycophenolic acid inhibits guanosine nucleotide synthesis and has been shown to be a potent inhibitor of lymphocyte proliferation as well as being effective at decreasing the incidence of graft rejection. Guanosine nucleosides are essential for protein glycosylation and many cell surface proteins including adhesion molecules, which are important for graft infiltration and rejection, are glycoproteins. There have been conflicting reports concerning the ability of MPA to interfere with glycosylation in lymphoid cells. Therefore, the purpose of this study was to investigate the effects of MPA on cell surface protein glycosylation in lymphoid cells. METHODS: Cells were cultured in the presence of increasing concentrations of MPA for different lengths of time and stained with fluorescent-labelled lectins specific for either mannose or fucose residues on glycoproteins. Analysis was then performed by flow cytometry. RESULTS: MPA treatment had no effect on the binding of either fucose or mannose-specific lectins to Con A stimulated human PBLs and rat lymph node lymphocytes or to a CEMC7a T cell line. CONCLUSION: The results show that, contrary to previous reports, MPA does not affect cell surface glycosylation in T cells using T cells from different sources of both human and non-human origin.

Equine placental cup cells show glycan expression distinct from that of both chorionic girdle progenitor cells and early allantochorionic trophoblast of the placenta.

Using lectin histochemistry on plastic-embedded material, the glycosylation patterns of equine girdle and cup cells, and associated endometrial glands, have been investigated from 37 to 67 days gestation. Results were compared with the glycosylation of the 50-day allantochorionic trophoblast of the established equine placenta that will later form the microcotyledons. The differentiated cup cells, which secrete equine chorionic gonadotropin (eCG), showed a pattern of glycosylation that was distinct both from the progenitor girdle cells and the allantochorionic trophoblast, with granules that bound lectins indicating high levels of alpha2,6 and alpha2,3-linked sialic acid, N-acetyllactosamine and bi/tri antennary non-bisected and bisected complex N-glycan. This is consistent with the known carbohydrate content of eCG. In contrast, the allantochorionic trophoblast at 50 days lacked detectable amounts of sialic acid and showed high levels of tri/tetra-antennary non-bisected complex N-glycan and N-acetyl galactosamine which was absent in the cup cells. During the process of girdle cell migration into maternal tissues, the uterine glands became greatly enlarged and dilated basally, with increased amounts of glycosylated secretory products revealed by lectins, which often seeped out into the extracellular space via ruptures in the apical regions of the gland wall.

A lectin binding analysis of glycosylation patterns during development of the equine placenta.

The glycosylation of the equine interhaemal barrier and areola was studied throughout the period of gestation. Placentae of 35, 37, 50, 119, 152, 200, 280 and 300 days gestation were investigated, using semithin plastic embedded sections and a panel of 15 biotinylated lectins with an avidin-peroxidase revealing system. Glycosylation of the trophoblast and maternal epithelium showed the most change during the first 50 days of gestation, being associated with the initial stages of adhesion and attachment. In the trophoblast, non-bisected tri/tetraantennary complex N-glycan was only evident after day 37 and terminal N-acetyl galactosamine, alpha2,3- and alpha2,6-linked sialic acids disappeared at the same time. The areolar trophoblast exhibited some differences from microcotyledonary areas, especially with respect to 2-deoxy, 2-acetamido alpha-galactose and tri/tetraantennary, non-bisected complex N-glycan, suggesting that the differences in function between microcotyledonary and areolar trophoblast are reflected at both the morphological and the biochemical level. Granules of the maternal uterine epithelium bound many lectins, particularly those with specificity for bisected and non-bisected bi/triantennary N-linked glycan, 2-deoxy, 2-acetamido alpha-galactosyl, beta-galactosyl and some fucosylated termini. Binding to sialic acids in alpha2,3- and alpha2,6-linkage was sparse. Maternal and fetal capillaries showed little change in glycan expression over the period studied, being rich in bisected and non-bisected bi/triantennary N-linked glycan and sialic acids, with some terminal N-acetyl galactosamine and no detectable terminal fucosyl residues.

Expression of proteoglycans by cultured chick sternal chondrocytes.

Chondrocytes isolated from the caudal and cephalic ends of the sterna of embryonic chicks were cultured in collagen gels. Differences were found, histochemically, between the proteoglycans produced by the "caudal" and "cephalic" cultures and with length of time in culture. The cultures were labelled with [14C]galactose and [35S]sulphate at 7 and 21 days in culture and labelled compounds from media, and cell and matrix extracts analysed with Sepharose CL-2B. A large aggrecan-like proteoglycan was detected in the media with some aggregated proteoglycans found in the cell extracts even under the dissociating conditions used. One group of 14C-labelled compounds, found in the cell and matrix extracts, was equivalent in size to chick aggrecan core protein. Smaller proteoglycans and glycoprotein glycans were present. The types and proportions of these proteoglycans varied between the two cell types demonstrating biosynthetic commitment.

Lectin histochemistry and cytochemistry-light microscopy : avidin-biotin amplification on resin-embedded sections.

Within the last 25 yr, the study of glycans, their structures, and their drstribution in tissues has emerged from relative obscurity to become a major theme of molecular and cellular biology: "glycobiology" (1). Glycans are major components of cellular surfaces (2,3), extracellular matrices (4,5), and secretions (6), and play important roles in cell-cell and cell-matrix recognition and adhesion (7,8). They regulate the surface environment of cells by influencing the structure of water (6,9), by modulating diffusion, by sequestering metabohtes such as metal ions, by presenting various growth factors to their receptors (10,11), by acting as ligands in recognition-adhesion systems (11-13), and by making major contributions to cell surface charge (2,14). In secretions, they are variously determinants of molecular folding (15), hydration, interaction, and targeting, and they serve in the mechanisms that monitor the aging of glycoconjugates in circulation (6,9,16,17). They are now implicated in mechanisms of molecular targeting and segregation within the endomembrane systems of cells (18-20), in calcium transport in mitochondria (21), the handling of mRNA and its export from the nucleus, the regulation of transcription (19), and in then classical role as energy stores. In all of this, the association of anatomical or ultrastructural localization with biological function is of paramount importance. specific glycans occur in specific places (22,23). The challenge for the histochemist is to reconcile the achievement of sufficiently precise anatomical localization of glycans with the maximum of chemical information about their nature. The lectins have been the main means of accompllshing this, since their first application as fluorochrome-labeled probes to paraffin sections in the early 1970s (2,24). More recently, fluorescence has been largely supplanted by nonfluorescent disclosing systems, such as biotin-avidin-peroxidase and, though paraffin sections are still widely used, resin-embedding of specimens has proved advantageous in terms of resolution and economy of material at the light microscopic level.

Environmental pollutants as aetiological agents in female reproductive pathology: placental glycan expression in normal and polychlorinated biphenyl (PCB)-exposed mink (Mustela vison).

Polychlorinated biphenyls (PCB) may cause growth retardation or fetal death in mink. Pathological changes in endotheliochorial mink placentae were examined following exposure to PCB during gestation. Placentae from six animals with average fetal crown-rump (C-R) lengths between 16 and 53 mm given 0.65 mg/day Clophen A50 (low dose), and one from five animals with an average fetal C-R length of 14 mm given 1.3 mg/day (high dose), were examined. Mink were treated from 9 to 24 days before mating until killed at day 53. Placentae were formalin-fixed with four size-matched controls and embedded in resin. Sections were stained with five biotinylated lectins to detect specific glycans. Both control and treated (low dose) mink showed degenerative changes in maternal endothelium from 13-16 mm, revealed by increased lectin binding, caused probably by high cell turnover during tissue remodelling. Controls of 47 and 50 mm exhibited fewer degenerate maternal endothelial cells. The 31-mm PCB-treated tissue showed separation of the trophoblast from the interstitial layer and, at 53 mm, loss of its normal architecture, increased damage to maternal endothelium and infarction. High-dose PCB was extremely toxic, producing fetal death or extensive placental infarction by 14 mm C-R length. Lectin staining thus revealed the effects of PCB toxicity, shown by increased injury to maternal endothelium and severe trophoblastic damage.

Localisation of glycans in the placenta: a comparative study of epitheliochorial, endotheliochorial, and haemomonochorial placentation.

Specimens of mid-term (horse), near-term (pig, cow, sheep, mink) and term (human) placentae and associated tissues have been examined with a panel of 15 biotinylated lectins combined with an avidin-peroxidase revealing system. The aim of this study has been to analyse the expression of glycans at the materno-fetal interface in order to establish whether the morphological diversity exhibited by these six species is reflected by accompanying biochemical diversity, or whether similar types of glycan are expressed in tissues performing similar functions. Lectin staining intensity was scored in the following elements of the interhaemal placental barrier: maternal capillaries, maternal uterine epithelium, the materno-fetal interdigitating microvillous membrane (brush border in the human), trophoblast, and fetal capillaries. A high degree of biochemical diversity was found in the glycan expression of the various placental components within and among placental types. Each layer showed widely differing patterns of lectin binding between species, with only a few findings in common: 1) the relative lack of simple fucosyl termini, 2) the presence of non-bisected bi/tri-antennary N-glycan in most layers, 3) an abundance of terminal N-acetyl galactosamine, and 4) the restriction of high mannose glycans to intracellular granules. This diversity may be a mechanism to avoid hybridisation, although glycan patterns may change between conception and placental development, or it may have evolved as a consequence of morphological changes. It is possible that it may also be part of the cause, rather than the result, of the structural diversity that is so characteristic of mammalian placentation.

South Asians with ulcerative colitis exhibit altered lectin binding compared with matched European cases.

Ulcerative colitis is associated with abnormalities of mucin synthesis and secretion, features that may also be associated with malignant change. It has been shown that South Asians in Britain have a high incidence of ulcerative colitis but a low incidence of colorectal carcinoma compared with their European counterparts. Previous studies have demonstrated changes in colonic mucin sialylation and sulphation in both South Asian and European cases with ulcerative colitis. This was related to disease severity, but changes were also found in quiescent disease. The aim of the present study was to determine glycoconjugate expression in the colon from South Asian cases and to compare results with those from a group of affected Europeans. Glycans were identified in formalin-fixed, paraffin-embedded tissue from 17 South Asian patients with ulcerative colitis and from 11 European patients with a similar degree of colitis, by the application of 10 biotinylated lectins. These were directed against a range of sialyl, fucosyl and 2-deoxy, 2-acetamido-galactosyl sequences, using an avidin-peroxidase revealing system and semiquantitative assessment. The South Asian group showed a reduction in the binding of agglutinins from Sambucus nigra in the apical-membranous region of enterocytes, and a decrease in apical Maackia amurensis agglutinin binding. These results suggest that South Asians with ulcerative colitis show a different distribution of terminal N-acetyl neuraminyl residues, either in their alpha-2,6 or alpha-2,3 linkage, compared with their European counterparts. The changes in sialylation observed in European cases compared with normal disease-free control subjects were present in quiescent disease, but were also related to disease activity. Their absence in Asians with ulcerative colitis may imply an inherent, genetically determined variation in this group, which may also play a part in their reduced risk of subsequent malignancy.

A comparative study of lectin binding to cultured chick sternal chondrocytes and intact chick sternum.

Cultured chondrocytes derived from the caudal and cephalic ends of embryonic chick sterna have been compared with each other and with whole sternum, by using a panel of 21 lectins to probe the distribution of oligosaccharides in glycoconjugates of cells and matrix at various times of culture or development. On culture in collagen gels, the cells changed their morphology with time, degrading glycan in the surrounding culture medium and depositing new matrix, the glycan content of which reflected the site of origin of the cells, indicating that the glycan phenotype of both cells and matrix ('glycotype') was predetermined and persistent. Sterna of embryonic chicks showed unexpected complexity in their distribution pattern of glycan, containing at least six distinct regions. Major regional temporal differences were evident among saccharides terminating in alpha-N-acetyl galactosamine and beta-galactose, while changes in glycans terminating in fucose, sialic acid and alpha-mannose were somewhat less marked. Subsets of complex N-glycans changed little.

Analysis of an HIV-infected cohort followed for as long as 15 years after seroconversion.

Data from a cohort of 62 HIV-positive individuals with hemophilia or von Willebrands disease infected for a maximum period of 15 years were analyzed. The relation between CD4+ and total lymphocyte counts and their rate of decline was analyzed with respect to age at seroconversion, time of seroconversion, and development of disease and subsequent death. As expected, the CD4+ and total lymphocyte population decline correlated with increased probability of disease and death. The patients fell into two distinct categories with respect to this decline: those whose cell count declined steadily (single slope) and those whose cell count remained steady or decreased very slowly for a variable period and then declined sharply (double slope). Within this cohort, the presence of a double slope appears to indicate a poorer prognosis, as 9 of 18 of the patients who have died showed this pattern, whereas only 6 of 42 of the remaining patients have this pattern even though more than half of this group have CD4+ lymphocyte counts < 0.2 x 10(9)/L. In addition, the ratio of CD4+ lymphocyte count to total lymphocyte count decreased with increasing cumulative frequency of the cumulative incidence of disease and death and the overall probability of death in this cohort was lower than expected, being 30% 12 years after seroconversion.

Glycans of the trabecular meshwork in primary open angle glaucoma.

AIMS: Glycan expression was compared in glaucomatous trabecular meshwork (TM) and normal TM in order to determine any differences which may reflect pathological changes underlying primary open angle glaucoma (POAG). METHODS: Resin embedded TM from trabeculectomy specimens from 15 eyes with POAG and from 12 eyes with normal anterior segments were probed with a panel of biotinylated lectins and an avidin-peroxidase revealing system at the light microscope level. Statistical analyses were performed on the comparative staining results. RESULTS: The lectins ConA and ePHA showed strong staining in all areas of both glaucomatous and normal TM; ePHA staining of Schlemm's canal (SC) from POAG TM was significantly less than that from normal TM (ePHA-SC p = 0.04). The lectins PSA, LCA, and SNA bound moderately strongly to SC endothelium and weakly to the endothelium of the corneoscleral meshwork (CSM); glaucomatous SC endothelial binding was significantly less than that of normal SC endothelium for PSA and LCA (PSA-SC p = 0.002, LCA-SC p = 0.002). STA and DSA showed moderately strong binding while WGA, ECA, AHA, and MPA bound weakly throughout the TM; for DSA and MPA this staining was significantly greater in POAG than in normal TM (DSA-SC p = 0.001, DSA-CSM p = 0.002, MPA-SC p = 0.01, MPA-CSM p = 0.02). Jac stained strongly throughout the TM and showed no significant difference in POAG compared with normal TM (Jac-SC p = 0.6, Jac-CSM p = 1). 1PHA, SBA, DBA, CTA, UEA-1 and LTA did not bind to glaucomatous TM or normal TM. There were no age-related changes seen. CONCLUSIONS: The expression of some complex and hybrid, bisected and non-bisected N-linked glycans is significantly diminished in glaucomatous TM compared with normal TM. Some glycans with multiple N-acetylglucosamine residues and O-linked glycans with terminal and subterminal galactosyl groups are significantly increased in POAG TM. Glycan expression does not change significantly with age in POAG or normal TM.

Glycoconjugates of the human trabecular meshwork: a lectin histochemical study.

Twelve specimens of resin-embedded human trabecular meshwork were probed with a panel of 21 biotinylated lectins, using an avidin-biotin peroxidase revealing system, in order to determine the normal pattern of saccharide expression in this tissue. High-mannose, intermediate and hybrid N-linked glycans, and complex N-linked bisected and non-bisected bi/tri-antennate glycans, as shown by the binding of Canavalia ensiformis (ConA), Pisum sativum (PSA), Lens culinaris (LCA) agglutinins and Phaseolus vulgaris erythroagglutinin (ePHA), were strongly expressed by the canal of Schlemm endothelium and juxtacanalicular tissue, but less so by the corneoscleal meshwork. Highly branced complex glycans were not found, as there was no binding by Phaseolus vulgaris leukoagglutinin (IPHA). Sialyl residues, especially those alpha 2,6-linked as demonstrated by strong Sambucus nigra (SNA) lectin staining, were also abundant in this area. N-acetyllactosamine sequences and some O-linked glycans were present in the trabecular meshwork, as shown by Solanum tuberosum (STA), Datura stramonium (DSA), and Jacalin (Jac) lectin binding, while fucose residues were not detected by Tetragonolobus purpureas (LTA) or Ulex europaeus-1 (UEA-1) agglutinins. These results indicate similarities with renal glomerular and vascular endothelium, although the lack of binding with UEA-1 agglutinin suggests differences which may relate to the specialized function of the trabecular meshwork. This study provides a baseline for comparative analysis of the glycans of human trabecular meshwork in pathological conditions such as primary open-angle glaucoma.

Glycans of the early human yolk sac.

The pattern of glycan distribution in the early human yolk sac has been investigated using a panel of lectins. Two 6-week and one 8-week human yolk sacs, and one 8-week fetal liver from live, ectopic pregnancies were fixed and embedded in epoxy resin. Lectin histochemistry was carried out on sections of these tissues using 23 biotinylated lectins and an avidin-biotin peroxidase revealing system. Mesothelial surfaces expressed most subsets of N-glycans (other than high mannose types), N-acetyl-lactosamine, sialic acid, and alpha 1,6-N-acetylgalactosamine. Endodermal surface and lateral membranes resembled those of mesothelium, but showed a preponderance of alpha 2,6-sialyl residues. Most intracellular granules contained N-glycan. There was a marked heterogeneity of granules in the endodermal cells, with different subsets varying in both staining and positional characteristics. The mesenchymal matrix bound most of the lectins used in the study, and expressed fucosyl residues which were also detected in the endothelium. Fetal liver parenchyma showed very similar staining patterns to those seen in the endoderm except for the distribution of N-acetylglucosamine, which was sparse. Despite some common features, each germ cell layer had a distinct 'glycotype', with some saccharides showing extreme topographical restriction.