Lectin histochemistry of galactose and N-acetyl-galactosamine glycoconjugates in normal gastric mucosa and gastric cancer and the relationship with ABO and secretor status

The histochemical binding of four lectin-peroxidase conjugates to normal human gastric mucosa and gastric carcinoma is described. The lectins were peanut agglutinin (PNA) which is specific for galactose residues and soy bean agglutinin (SBA), Dolichos biflorus agglutinin (DBA) and Helix pomatia agglutinin (HPA) which are specific for N-acetylgalactosamine. Binding of PNA to surface mucous cells or normal gastric mucosa occurred in non-secretors but not secretors and was independent of ABO blood group at all sites. PNA binding was unrelated to the immunohistochemical demonstration of Thomsen-Friedenreich (T) antigen. DBA and HPA bound selectively to surface mucous cells in normal gastric mucosa from group A secretors but binding at other sites was independent of ABO status. SBA binding showed no relationship with blood group or secretor status. In gastric cancers the major finding was the occurrence of extensive masking of lectin binding sites by sialic acid which was not seen in normal mucosa. Sialic acid masking was most marked with PNA and least marked with DBA. There was no correlation between lectin binding patterns and the stage or differentiation of tumours. Results are consistent with in vitro studies demonstrating increased sialation of membrane glycoproteins following malignant transformation. Difficulties in interpreting the histochemical demonstration of lectin binding in terms of specific glycoconjugates are discussed.     

Histochemical study of glycoconjugates in active and photoperiodically-regressed testis of hamster (Mesocricetus auratus)

The objective of the present study was to characterize glycoconjugates of hamster testis in gonadally-active and -inactive states by lectin histochemical methods. Thirteen HRP- or digoxigenin-labeled lectins were used in samples obtained from fertile and photoinhibited hamsters. In gonadally-active hamsters, spermatozoa tails were stained with Con-A, HPA, PNA, UEA-I, LTA, AAA, WGA and LFA and weakly with GNA and RCA-I. Spermatozoa acrosomes were labeled with HPA, SBA, WGA and PNA. Spermatid acrosomes were labeled with SBA, RCA-I, PNA, and WGA. Staining with GNA and Con-A was found in the Golgi phase and HPA staining was found in the Golgi phase and maturated spermatids. Cytoplasm of spermatocytes was labeled with Con-A, GNA, LTA, AAA, RCA-I, HPA, WGA and LFA, whereas spermatocyte membranes were stained with Con-A, LTA and AAA. Spermatogonia were strongly labeled with Con-A and moderately labeled with AAA, WGA and LFA. Sertoli cells were positive after staining with Con-A, AAA, WGA, and LFA. The lamina propria was positive after staining with UEA-I, LTA, AAA and LFA. Leydig cells showed strong labeling with SBA, Con-A, GNA, SNA and MAA, moderate labeling with WGA, weak labeling with RCA-I, AAA and LFA. In gonadally-inactive hamsters, spermatocytes showed increased staining with HPA, PNA and AAA, whereas staining with Con-A, GNA and LTA had disappeared. Spermatogonia showed an increased labeling with AAA and WGA, but labeling with Con-A and LFA had disappeared. Sertoli cells were strongly labeled with GNA. Con-A and GNA staining was decreased in Leydig cells of gonadally-inactive hamsters but PNA and HPA staining was increased. The lamina propria in regressed testes showed intense labeling with PNA. These results suggest that histological, morphological and hormonal changes occurring in hamster testis during exposure to a short photoperiod are reflected in altered patterns of expression and distribution of N- and O-linked glycans.     

GP-83 and GP-39, two glycoproteins secreted by human epididymis are conjugated to spermatozoa during maturation

Surface glycoconjugates of spermatozoa are modified during epididymal maturation, which is closely related to the development of sperm function. In addition, recognition of surface glycoconjugates is one of very critical events in sperm-oocyte interaction. The binding of carbohydrate-specific lectins to the human sperm surface during epididymal maturation has been investigated. However, the glycoproteins responsible for lectin binding in sperm maturation are not well documented. This study used wheat germ agglutinin (WGA), peanut agglutinin (PNA) and concanavalin A (Con-A) to identify sperm maturation-related glycoproteins in human epididymis. Histochemical localization revealed that the binding sites of WGA, PNA and Con-A were mainly in the principal cells and luminal contents of the human epididymis, but not in the interstitial regions. Each lectin displayed a fairly distinct regional localization. On Western blots probed with WGA and Con-A, glycoproteins of 83 kDa (GP-83) and 39 kDa (GP-39) were identified in the sperm extracts, epididymal fluid and tissue extracts of the corpus and cauda epididymides, but not in the caput. PNA identified GP-83 in the same manner as WGA and Con-A, but did not recognize GP-39. These results suggest that lectin-binding glycoproteins GP-83 and GP-39 found on mature spermatozoa may be secreted by the principal cells of corpus and cauda epididymis, and conjugated to spermatozoa during their transit in human epididymis.     

Light and electron microscopic studies of lectin binding on the glycocalyx of rat pancreatic cells. I. Normal tissue and isolated cells.

Lectin binding of the glycocalyx of pancreatic tissue sections as well as of isolated pancreatic acini and acinar cells was studied of healthy wistar rats by light and electron microscopy. For light microscopy, we used FITC (WGA, RCAI, LCA) or peroxidase marked (WGA, RCAI, PNA, PHA, LCA, UEAI, LPA) as well as unmarked lectins (Con A, VAA I). Gold marked lectins were used for electron microscopy (WGA, RCAI, LCA, HPA, PNA, VAAI-B). Intact acinar cells in pancreatic tissue sections and isolated acini showed a strong binding of WGA, RACI, and HPA on the apical cell surface, whereas VAAI, UEAI, LCA, and Con A reacted strongly with the basolateral glycocalyx, but not with the apical surface. The 2 main domains of the glycocalyx of pancreatic cells showed their specific lectin binding so long as the junctional complexes between the cells are intact. The polarity of the cell surface of pancreatic acinar cells is discussed in regard to the possible function of the 2 domains.     

家兔附睾管尾段吸收功能的研究

家兔输精管低位结扎后附睾管尾段结构出现明显变化，外形和光电镜观察显示其管径增大，管腔扩张，腔内精子密集。附睾的平均重量增加。主细胞顶部胞膜向胞质内凹陷增多，并形成施工吞饮小泡，核上区出现许多与吸收作用有关的小泡、大泡、多泡体、溶酶体和线粒体等超微结构。出现上述变化的时间是术后的第３个月。输精管中位结扎组的变化情况基本相似于低位结扎组。输精高位结扎后因不能排出的睾网液可在附睾的近段被大部吸收，少量的     

Epididymis of viscacha (Lagostomus maximus maximus): morphological changes during the annual reproductive cycle

Little is known about morphological changes in the epididymis in relation to the natural photoperiod or their influence on sperm maturation. The viscacha is a seasonal rodent living in the Southern Hemisphere. The adult males exhibit an annual reproductive cycle with periods of maximum gonadal activity and gonadal regression. In this work, we studied seasonal variations in the morphology and cellular population of the epididymis during both periods, and we compared these results with those recorded at the testicular level. Epididymides were removed and studied by light microscopy. Measurements of luminal diameter, epithelial height, thickness of the lamina propria, and relative cellular distribution were performed. Analysis of variance (ANOVA) or nonparametric ANOVA was used to compare the results. Striking quantitative and qualitative changes were observed. Epididymides in periods of gonadal regression showed a significant decrease in luminal diameter and epithelial height in cauda, while the thickness of the lamina propria increased. In the epididymal corpus, the number of clear cells increased, and the cytoplasm of principal cells showed numerous giant vacuoles. During the active period, the number of halo cells increased and the cytoplasm of these cells was filled with dense bodies. In conclusion, the epididymis of the viscacha exhibits important seasonal morphological changes throughout annual reproductive cycle. The epididymal corpus and cauda segments appeared to be the segments most sensitive to seasonal cyclical variations of the external environment. We therefore postulate that the epididymal morphology of the viscacha probably could be regulated by the natural photoperiod.     

Lectin histochemistry of developing submandibular glands of fetal Syrian golden hamsters

Summary Lectin binding was studied in the developing submandibular glands of fetal Syrian golden hamsters (Mesocricetus auratus) from gestational day 12 to 16 (the day of birth). The fetuses were fixed, embedded in paraffin, sectioned and stained with nine lectin-horseradish peroxidase conjugates: concanavalin A (Con A), wheat germ agglutinin (WGA), Dolichos biflorus agglutinin (DBA), Helix pomatia agglutinin (HPA), Maclura pomifera agglutinin (MPA), Griffonia simplicifolia agglutinin I-B₄ (GSA I-B4), peanut agglutinin (PNA), Ulex europens agglutinin I (UEA I) and Limulus polyphemus agglutinin (LPA). The developing glands showed dramatic morphological alterations on a daily basis, accompanied by progressive changes in lectin staining. On day 12 the primitive gland showed only trace lectin staining with WGA, HPA, MPA, PNA and UEA I, but by day 13, strong staining with these lectins, as well as with DBA, was seen at the ductal lumenal surface, after the formation of the ductal lumens. Secretory granules first appeared in cells of the primitive acini on day 14; the secretion products were stained strongly with WGA, DBA, HPA, MPA, PNA and UEA I. On day 15, the secretion products were also stained moderately with GSA I-B4. Secretory differentiation was further developed on day 16, but the staining intensity of the mucins with the different lectins varied among the secretory cells. LPA failed to stain any part of the gland throughout the observation period, and Con A stained only glycogen.This work was supported by National Institutes of Health Grant HL37640.     

Lectin histochemistry on the dorsal epidermis of the Breton dog

Expression of sugar residues and the nature of oligosaccharide linkage during keratinocyte maturation in the epidermis of the Breton dog were studied with the use of lectin histochemistry. Thirteen lectins were used. Labelling was not observed with GSA I-B4, GSA II, UEA-I, and LTA. The cytoplasm of keratinocytes reacted with PNA, HPA, Con A, and WGA from the basal layer to the granular layer. PNA and Con A showed highest reactivity in the granular cell layer. The cell surface showed increased reactivity with PNA, HPA, and WGA with maturation of keratinocytes. KOH-neuraminidase treatment (KOH-Neu) increased PNA and RCA120 staining during keratinocyte differentiation thus indicating an increase in oligosaccharides terminating with sialic acid-Galbeta(1,3)GalNAc and sialic acid-Galbeta(1,4)GlcNAc, respectively. Labelling of the glycocalyx of basal and spinous keratinocytes with SNA and MAA revealed terminal Neu5acalpha(2,6)Gal/GalNAc and Neu5acalpha(2,3)Galbeta(1,4)GlcNAc. KOH-Neu-DBA showed oligosaccharides terminating with sialic acid-GalNAcalpha(1,3)GalNAc in the spinous and granular layers. A selective glycocalyx labelling of granular keratinocytes was observed with DBA and SBA. Reactions with MAA, PNA, DBA, RCA120, SBA, HPA, and WGA disappeared after the beta-elimination reaction. Our findings indicate that Breton dog epidermis contains more O-linked than N-linked oligosaccharides and confirm that different subpopulations of keratinocytes can be distinguished by lectin histochemistry.     

Glomerulocystic kidney disease in a Belgian Malinois dog: an ultrastructural, immunohistochemical, and lectin-binding study

Renal cysts in the cortex of a juvenile Belgian Malinois dog with acute renal failure were studied by means of light, scanning and transmission electron microscopy, immunohistochemistry for intermediate filaments, and binding for wheat germ agglutinin (WGA), peanut agglutinin (PNA), and Maclura pomifera agglutinin (MPA) lectins to determine the morphological and histochemical features of the epithelial cells of these cysts. The cysts were renal corpuscles with expanded urinary space. Glomerular tufts were small with poorly developed capillary loops and increased mesangial matrix. Continuity with the proximal tubule was evident in some cystic glomeruli. Two cell types lined Bowman's capsule. One was squamous with a central cilium and microvilli. The other had morphological and histochemical features of immature podocytes (parietal podocytes). These cells were round and protruded into the urinary space; they had thick cytoplasmic projections that resembled foot processes of podocytes, microvilli, and filtration slits. The parietal podocytes expressed vimentin and cytokeratins and had affinity for WGA as do normal immature podocytes. These features suggest that the parietal podocytes are derived by metaplasia of the parietal cells. The basement membrane of Bowman's capsule was irregularly thickened and showed multifocal glycosylation changes with lectin histochemistry (WGA, PNA, MPA) in areas adjacent to the parietal podocytes. Histologic and ultrastructural findings in this dog are consistent with glomerulocystic kidney disease. This is the second report of canine glomerulocystic kidney disease. Features are similar to those of the human counterpart, but it is unclear whether genetic defects cause the disease in the dog. The presence of parietal podocytes in all cysts suggests that abnormal differentiation may play an important role in the pathogenesis of this type of polycystic kidney disease.     

Lectinohistochemistry of human bladder cancer: loss of lectin binding structures in invasive carcinomas

With the purpose of studying changes in the expression of glycoconjugate structures in urothelium, nine different lectins (PNA, WGA, VFA, GSA II, STA, UEA I, LCA, DBA and HPA) with specificity for mono- or oligo-saccharides were used on formalin-fixed, paraffin-embedded tissue sections from 47 patients who had undergone surgical resection for bladder tumors and on normal urothelial biopsies from 10 patients. The tumors were graded and a lectinohistochemical method using biotinylated lectins and avidin-biotin-peroxidase complex was used to demonstrate the lectin binding. Positive staining reactions of cells in cytoplasm and on membranes were evaluated in the basal, the intermediate, and the luminal cell layers, respectively. In both normal and atypical urothelium lectin binding predominated in the luminal cell layer and decreased towards the basal cell layer. In normal urothelium all lectins stained greater than 66% of the cells in the luminal cell layer in cytoplasm and between 5 and 100% of the cells on membranes depending on the lectin used. A gradual loss of lectin-binding structures was seen with increasing grade of atypia. The range of this decrease varied considerably from one lectin to another, but it was consistently found that the percentage of cells stained in cytoplasm and on membranes decreased. A significantly lower percentage of cells stained in cytoplasm was found in invasive tumor cell-islands compared to normal urothelium. In invasive tumor cell-islands staining of cells on membranes was completely absent, except for HPA lectin that stained less than 10% of the cells. In conclusion, we demonstrate a dramatic decrease in lectin-binding carbohydrate structures associated with urothelial malignant progression.     

Lectin-binding patterns of small lymphocytes in bone marrow,thymus and spleen: demonstration of lymphocyte subsets by quantitative radioautography

Cells from mouse bone marrow, thymus and spleen were exposed to ¹²⁵I-labeled concanavalin A (Con A), Lens culinaris lectin (LCL), soybean agglutinin (SBA), Helix pomatia agglutinin (HPA), phytohemagglutinin-P (PHA-P), peanut agglutinin (PNA), or wheat germ agglutinin (WGA) in a range of concentrations and examined radioautographically. Small lymphocytes in the three organs differed in the minimal concentration of each lectin which gave detectable surface labeling, while at optimal lectin concentrations, their labeling intensity profiles differed markedly. Inhibition by sugars demonstrated the labeling specificity. Major populations of bone marrow small lymphocytes bound WGA strongly, while Con A, SBA, HPA, PHA-P and LCL were bound only weakly, and PNA binding was lacking. Most thymus cells bound Con A, SBA, HPA, PHA-P and PNA strongly, WGA and LCL weakly. Subsets of bone marrow and thymus small lymphocytes differed from the major populations in their lectin-binding intensities. Spleen small lymphocytes were heterogeneous in the binding of each lectin. However, a major population bound LCL exceptionally strongly, while few cells bound PNA. Using a panel of lectins under standardized conditions, these studies show distinctive lectin-binding patterns for small lymphocytes in the bone marrow, thymus and spleen, respectively. Major and minor cell populations are distinguishable in each organ, providing an approach to discriminating lymphocyte lineages, subtypes and differentiation stages.     

Ultrastructural Localization of Helix pomatia Agglutinin (HPA)-Binding Sites in Human Breast Cancer Cell Lines and Characterization of HPA-Binding Glycoproteins by Western Blotting

Ultrastructurally, cells of five human breast cancer cell lines (MCF7, BT549, BT20, T47D, and HBL100) generally displayed many characteristics of their epithelial origin. The most distinctive features were observed in MCF7 cells, which consistently showed microvilli and submembranous granules. These ultrastructural findings served as a basis for localizing the binding sites of the lectin Helix pomatia agglutinin (HPA). After use of the pre-embedding method consistent HPA labeling of the cell membrane was obtained in all the cell lines, and in association with microvilli and submembranous granules in the MCF7 cells. The HBL100 cells were not labeled by HPA irrespective of the method used. In addition, lysates from these cell lines were subjected to polyacrylamide gel electrophoresis and Western blotting with HPA to analyze these binding sites further. In the Western blots, however, lysates of HBL100 cells, in common with all those from the other cell lines, revealed a number of HPA-reactive bands, indicating the greater sensitivity of Western blots compared with the histochemical preparation. The principal band was of approximately 90 kDa, and it was suggested that this could be related to the transferrin receptor.     

Lectin binding pattern in human retinal pigment epithelium.

A comparative lectin histochemical study of human retinal pigment epithelium (RPE) was performed to investigate the lectin binding pattern of normal, reactive and proliferating RPE. Normal RPE with attached sensory retina was found to bind the lectins Con A, WGA, PNA and RCA I. Reactive and proliferating RPE in retinal detachment and in photocoagulation scars revealed the same lectin binding pattern although its cellular topography changed. RPE-macrophages showed an additional reaction with SBA. In periretinal membranes of human PVR the typical lectin binding pattern of Con A, WGA, PNA and RCA I was found in pigmented and in a subpopulation of non-pigmented cells, suggesting that these lectin-positive elements were of RPE-origin. Additionally, single pigmented cells positive for SBA were found indicating macrophage differentiation. Thus lectin histochemistry provides a tool for cytochemical identification of RPE and its morphologic variants by revealing a specific combination of sugar-binding sites.     

Changes in glycoconjugates revealed by lectin staining in the developing airways of Syrian golden hamsters.

Lectin binding was studied in the developing airways of Syrian golden hamsters on gestational days 11-16 (day 16 is the day of birth). The trachea and lungs were fixed in 4% formaldehyde-1% glutaraldehyde, 6% mercuric chloride-1% sodium acetate-0.1% glutaraldehyde, and 95% ethanol; embedded in paraffin; and stained with eight lectin-horseradish peroxidase conjugates: Triticum vulgare (WGA), Dolichos biflorus (DBA), Helix pomatia (HPA), Maclura pomifera (MPA), Griffonia simplicifolia I-B4 (GSA I-B4), Arachis hypogaea (PNA), Ulex europeus I (UEA I), and Limulus polyphemus (LPA). Each lectin yielded a characteristic staining pattern, which modulated throughout development. In general, changes in staining characteristics of the tracheal epithelium preceded similar changes in the lobar bronchus, bronchiole, and alveolus. In the case of UEA I, MPA, WGA, and HPA, staining increased with time uniformly over the luminal surface of all epithelial cells. However, in the case of PNA, GSA I-B4, and LPA, after the differentiation of ciliated and secretory cells, the apical surfaces of the ciliated cells stained more intensely than the apical surfaces of the secretory cells. Neuraminidase pretreatment enhanced PNA and GSA I-B4 staining in both cell types. In the case of PNA, these light microscopic observations were confirmed by ultrastructural study. Unlike the other lectins, the pattern of staining with DBA was unusual. Staining was moderate at first, then decreased (days 13 and 14), then increased at all airway levels. This study shows that different glycoconjugates modulate in airway epithelial cells throughout fetal development.     

Binding studies of gold labelled lectins on carbohydrate compounds of the flask cells in claw-frog kidney.

The carbohydrate compounds of the mucus of flask cells in the kidney of claw-frogs (Xenopus laevis) were studied by gold marked lectins (WGA, RCA, L, LCA, HPA, PNA). We used a post-embedding technique. Seminthin or ultrathin sections of Lowieryl K H M-embedded kidney tissue were incubated. For light microscopy, a gold-silver technique was used. The mucus of the flask cells reacted strongly with WGA, RCA L and HPA, whereas LCA and PNA showed no binding. The Golgi apparatus and small cytoplasmatic vesicles reacted also positively with WGA, RCA L and HPA. The autoradiographically detected secretion routes of the glycosaminoglycan-rich secretion of flask cells are also demonstrable by lectins.     

Lectin binding sites on human sperm and spermatogenic cells

Testes of sexually mature men were studied histochemically with 20 fluorescein isothiocyanate-labeled lectins. Based on their pattern of reactivity with intratesticular spermatogenic cells, lectins were divided into five groups: 1) lectins reacting with all spermatogenic cells (Suc. ConA, WGA, LCA, PHA-E, PHA-L, STA, MPA, and RCA-II); 2) lectin reacting with spermatocytes, spermatids, and spermatozoa, but not with spermatogonia (RCA-I); 3) lectins reacting with spermatids and spermatozoa only (BPA, PNA, SBA, and VVA); 4) lectins reacting only with spermatozoa (HPA, GSA-I, UEA-II, and GSA-II); and 5) lectins with no distinct staining of spermatogenic cells (DBA, LBA, and UEA-I). All lectins from groups 1-4 were reactive with ejaculated spermatozoa. On the basis of the staining patterns of the head region of ejaculated spermatozoa, four lectin reactivity groups were defined: 1) lectins reacting with the plasma membrane of the whole head (BPA, WGA, LCA, STA, RCA-II, PHA-E, PHA-L, RCA-I, UEA-II, and GSA-II); 2) lectin reacting with the acrosomal cap and postacrosomal region of the plasma membrane (Suc. ConA); 3) lectin reacting with the acrosomal cap region of the plasma membrane (PNA); and 4) lectins reacting with the midregion of the sperm head in a bandlike manner (HPA, VVA, SBA, GSA-I, and MPA). These data provide a map of lectin binding sites on human testicular spermatogenic cells and ejaculated spermatozoa and show that the distribution of glycoconjugate domains of spermatogenic cell changes during differentiation and maturation.     

The glycosylation of thymic microenvironments. A microscopic study using plant lectins

The thymus is the principal organ for development of T-cells. Thymocyte precursors from bone marrow-derived progenitor cells enter the thymus where they differentiate involving several differentiation stages into mature T-cells that can leave the thymus to the periphery. Migration of thymocytes through the thymus and their development are tightly controlled by the interaction of thymocytes with components of the thymic microenvironments. Several studies have demonstrated the pivotal importance of glycosylation in cell-cell interactions or interactions of cells with extracellular matrix components (ECM) in various physiologic processes in the body. The knowledge on glycosylation of thymic microenvironments is however limited although the presence of C-type lectin receptors such as DC-SIGN, mannose receptor and DEC-205, which are specifically recognizing distinct carbohydrate moieties emphasize the importance of glycosylation in the thymus. In order to outline the distribution of glycoconjugates in microenvironments of the human thymus we studied the glycosylation of the human thymic microarchitecture by using plant lectins in situ. Eleven plant lectin-biotin conjugates with distinct specificity were used and analyzed by fluorescence microscopy. Mannose glycoconjugates, specifically detected by the lectins GNA and NPA, were abundant in the cortex but not in the medulla. Dendritic cells present in the thymic cortex were specifically co-stained with the galactose-specific lectins DSA and PNA. Several lectins bound to the thymic vasculature. The alpha2-fucose-specific lectin UEA stained thymic blood vessels in the interlobular space and medulla and capillaries in the cortex. In addition to UEA, thymic blood vessels and capillaries also reacted with the lectins DSA, PNA and the alpha-GalNac-specific lectin HPA. In contrast, lymph vessels present in the interlobular space do not interact with UEA, DSA and PNA, but only with HPA, revealing a disparate glycosylation pattern of lymph and blood vessels that may be important to determine the direction of thymocytes entering or leaving the thymus. In conclusion, the restricted expression patterns of carbohydrates defined microenvironments in the human thymus highlight the importance of glycosylation in various steps of T-cell development.     

Peanut lectin binding to the alveolar lining layer in hyaline membrane disease

Summary The authors have studied the histochemical pattern of Peanut lectin (PNA) binding sites in lungs of seven newborns with hyaline membrane disease (HMD) and ten controls. The alveolar lining layer was positive in HMD and no changes in the PNA pattern was noted after neuraminidase digestion. Hyaline membranes were generally unstained but occasional reactivity was encountered in some parts. No reaction with PNA was observed in control lungs, but positivity was seen after neuraminidase pretreatment. Our histochemical data document the presence of accessible galactosyl residues with absence of terminal sialic acid in the alveolar lining layer of newborns with HMD. The authors suggest that PNA reactivity in HMD reflects an histochemical feature described in fetal lungs at the pseudoglandular and canalicular stages.This work was supported by grants from MPI Rome     

Lectin cytochemistry reveals differences between hamster trachea and bronchus in the composition of epithelial surface glycoconjugates and in the response of secretory cells to neutrophil elastase

Hamsters exposed to an intratracheal instillation of human neutrophil elastase (HNE) accumulate an abnormally high number of secretory granules in bronchial but not tracheal epithelial cells. We employed lectin cytochemistry to investigate possible differences in the epithelial cell surface glycoconjugate layer in trachea compared to bronchus which might explain the regional dissimilarity in response to HNE. Portions of glutaraldehyde-fixed trachea and bronchi were incubated in one of several ferritin-labeled lectins prior to embedding for transmission electron microscopy. Lectins from Ricinus communis, Helix pomatia, and Triticum vulgaris bound to the surface of tracheal secretory cells in moderate to profuse amounts, while most bronchial secretory cells showed little or no label with these lectins. Gold-labeled Helix pomatia agglutinin (HPA), a lectin specific for secretory cells, showed a decrease in surface binding to all tracheal secretory cell types within 2 h of HNE instillation, compared to saline controls. In contrast, the majority of bronchial secretory cells showed an HNE-induced increase in surface label from extremely low levels in saline controls. The low levels of lectin binding to bronchial cells, in contrast to the trachea, may indicate the lack of a protective surface glycoconjugate coat, thus explaining the vulnerability of these cells to HNE. The rise in number of accessible HPA binding sites on the surface of bronchial secretory cells exposed to HNE may represent an important event in the pathologic accumulation of secretory granules by these cells.     

Ultrastructure and lectin cytochemistry of the cloacal pelvic glands in the male newt Triturus marmoratus marmoratus

The cloacal organ of Salamandridae species contains four glands: pelvic, dorsal, ventral, and Kingsbury's glands. Pelvic glands have been studied only by light microscopy with conventional methods, and consist of multiple tubular serous glands with a prismatic epithelium which contains numerous PAS positive secretory granules. The present report is an ultrastructural and lectin cytochemistry characterization of the pelvic glands of Triturus marmoratus marmoratus throughout the reproductive cycle. Our methods consisted of conventional electron microscopy, and colloidal‐gold lectin cytochemistry of the following lectins: WGA, ConA, LcA, UEA‐I, PNA, SBA, and HPA. In the prereproductive period, the glands showed a tall epithelium which consisted of two cell types, dark and clear cells, surrounded by elongated, myoepithelial cells. Both dark and clear cells showed the ultrastructural characteristics of secretory cells, and exhibited many secretory granules in the apical cytoplasm. Areas showing densely packed, degenerating cell organelles—which were not surrounded by membrane—were observed in the dark cells whereas the clear cells showed large heterolysosomes. In the postreproductive period the number of secretory granules decreased, the rough endoplasmic reticulum was less developed, and areas of degenerating organelles were absent. In addition, small basal cells appeared. The results of the lectin histochemistry study were similar in both reproductive periods. In the epithelial cells, the rough endoplasmic reticulum, the Golgi complex, and secretory granules exclusively labeled to ConA. In all cell types, the nuclei reacted to all lectins while the cytosol only reacted to LcA lectin. The ultrastructural and histochemical characteristics of the pelvic glands of T. marmoratus suggest that these glands could be homologous to the mammalian seminal vesicles and prostate. Anat Rec 254:196–204, 1999. © 1999 Wiley‐Liss, Inc.