Griffonia Simplicifolia agglutinin-I Research Articles

Localization of Different Lectins and E-cadherin in the kidney of a Balady Egyptian Duck

The purpose of our present study is to determine the spatial pattern of different lectins and E-cadherin in the duck kidney, which has never been done before. We checked the localization of six different lectins in the adult duck kidney of both sexes by histochemistry using wheat germ agglutinin (WGA), ricinus communis agglutinin (RCA), peanut agglutinin (PNA), dolichos Biflorus Agglutinin (DBA), concanavalin (Con A) and griffonia simplicifolia agglutinin-I (GSA-I). We found different locations of lectins in the duck nephron and this difference may be related to the function of each lectin. WGA lectin had intense reactions in the proximal and distal convoluted tubules (PCT and DCT) but was not present in the renal corpuscle of the renal cortex. WGA was also intense in the medulla especially in the collecting ducts (CD) and collecting tubules (CT). RCA was intense in the renal corpuscle, moderate in in the renal cortex tubules, and intense in the medulla tubules. PNA was moderate in the tubules of renal cortex and renal corpuscle while in the medulla, PNA had intense reaction. Con A, GSA-I and DBA lectins had mild to moderate reactions in the renal cortex and also in the medulla. Con A and GSA-I had intense reactions in the renal corpuscles. E-cadherin, which is responsible for maintaining cell polarity, was located in the lateral epithelial cells of all tubes and ducts in the renal cortex and medulla and not present in the renal corpuscle. There are no previous studies describing the location of different lectins and E-cadherin in duck kidneys. Our study clarifies the distribution pattern of various lectins and E-cadherin, which may be useful for comprehending how these lectins and E-cadherin work in the duck kidney.

Apical tubules in marginal cells of the differentiating stria vascularis.

Apical tubules (ATs) in marginal cells (MCs) of the stria vascularis appear in limited stages of differentiation of the MCs, but their origin and roles remain uncertain. The present study was designed to solve the problem of whether the ATs are intracellular compartments derived from the Golgi apparatus (GA). The cochleae of Wistar rats at ages of postnatal days 1, 3, and 5 were prepared for electron microscopy and cytochemistry using thiamine pyrophosphatase (TPPase) and coenzyme A phosphatase (CoA-Pase) as marker enzymes of trans Golgi cisterns and fluorescent labelled lectin, griffonia simplicifolia agglutinin-I (GS-1). The ATs appeared in the apical cytoplasm of the MCs between postnatal days 1 and 5. Reaction products of TPPase and CoA-Pase activities were localized in the trans-Golgi cisterns and the ATs, which were occasionally in a close apposition to the GA. The reaction was found along the apical plasma membrane of the MCs only in case of TPPase. Heavy reactions to GS-1 were seen in the supranuclear region as well as along the apical plasma membrane of the MCs. The present ultrastructural and cytochemical studies indicate that the ATs, which appear in the MCs at limited perinatal stages, originate from the trans-Golgi cisterns. These ATs may be involved in the apical plasma membrane supply for the differentiation of the MCs prior to the generation of EP.

Genital lesions following long-term administration of clenbuterol in female pigs.

Pathologic findings, lectin histochemistry, and nuclear estrogen receptors were studied in the reproductive organs of gilts treated with clenbuterol. A ration containing 1 ppm of clenbuterol was fed for 40 days to four Landrace x Large white, 9-month-old gilts, weighing 134 to 172 kg at slaughter (gilt Nos. 5-8). Four gilts (Nos. 1-4) served as controls. Treated animals had macroscopic lesions characterized by microcystic ovaries and uterine atrophy. Histopathologic lesions included atretic degeneration of many ovarian follicles, complete absence of functional corpora lutea, a reduction in the number of endometrial glands, and a decrease in cytoplasmic volume of endometrial and glandular epithelial cells. In ovaries, uterus, and vagina lectin histochemistry, performed with thirteen different biotinylated lectins, revealed a different staining distribution between control and treated gilts. The binding pattern of Ricinus communis agglutinin-I (RCA-I) and -II (RCA-II) in the ovaries of control gilts, displayed labeling of cytoplasm in theca interna cells of Graafian follicles. There was no labeling of the same cells in treated gilts. Labeling patterns with Griffonia simplicifolia agglutinin-I (GS-I), Phaseolus vulgaris agglutinin (PHA), RCA-I and RCA-II documented a difference in the vascularity of the theca interna between Graafian follicles of control and treated gilts. The GS-I and Ulex europaeus agglutinin-I (UEA-I) binding patterns in uterus and vagina of treated gilts when compared to control gilts suggested that there was a block of the cycling activity in the proliferative stage. Immunohistochemical staining for estrogen receptors in the endometrium was positive in all but one treated gilts, and negative to weakly positive in control gilts. Serum progesterone concentrations were decreased in treated animals when compared to control; estradiol concentrations were similar in both group of gilts. Cystic ovaries, uterine atrophy, and reduction in progesterone concentrations suggested that clenbuterol changed ovarian hormonal activity in treated animals.

Lectin histochemical study of the kidney in uninephrectomized and diabetic (UN-D) mice.

Lectin histochemical characteristics of the kidney of experimentally induced diabetic mice were investigated. Mice were divided into the following 4 groups: control (C), streptozotocin (SZ)induced diabetes (D), unilateral nephrectomy (UN), and unilateral nephrectomy-SZ-induced diabetes (UN-D) groups, and killed at 12 weeks after completion of the SZ-injection. No lectin histochemical differences were observed in the glomeruli of all groups. Stainabilities of Bowman's capsules lined with cuboidal to columnar epithelia seen in C and UN groups were the same as those of proximal tubules, and they were different from those of Bowman's capsules lined with flattened epithelia generally seen in D and UN-D groups. In D and UN-D groups, distal tubules and collecting ducts increased their affinities to Canavalia ensiformis A, Dolichos bifiorus agglutinin, Griffonia simplicifolia agglutinin-I, Glycine max agglutinin, and Triticum vulgaris agglutinin, compared with C and UN groups. Conclusively, this study showed that unilateral nephrectomy enhanced SZ-induced kidney lesions without a prominent histochemical modification.

Lectin Histochemistry on Squamous Metaplasia in Different Epithelial Tumors of Dogs

Biotinylated lectins and avidin-biotin-peroxidase complex were used to study the correlation between cellular glycoconjugates' expression and squamous maturation in normal canine skin and in various epithelial neoplasms. Normal skin tissue was obtained from five, male, random-source dogs, 5 to 7 years old. The tumors tested, selected from the files of our Department, were fifteen squamous cell carcinomas from different tissue origin, five hepatoid perianal gland adenocarcinomas with squamous metaplasia, and fourteen solid mammary carcinomas with and without histologic evidence of squamous metaplasia. Except for mammary gland carcinomas, all tumors had been surgically excised from male dogs. Intermediate filament aggregation of twelve solid mammary gland carcinomas were studied electron microscopically. The basal and the lower spinous cells in normal skin and the less differentiated cells in squamous cell carcinomas stained moderately with Griffonia simplicifolia agglutinin-I. Spinous and granular cell layers stained strongly with Phytolacca americana mitogen and Arachis hypogaea agglutinin. Both lectins stained well-differentiated cells in squamous cell carcinomas. The electron microscopic study carried out in solid carcinomas of mammary glands revealed some relationship between the presence of intracytoplasmic tonofibrils and the binding of Griffonia simplicifolia agglutinin-I and Phytolacca americana mitogen to the tumors tested. Our results suggest that the glycosylation pattern occurring during normal keratinocyte differentiation is conserved in squamous cell carcinomas and that Griffonia simplicifolia agglutinin-I and Phytolacca americana mitogen may represent useful tools in distinguishing poorly differentiated squamous cell carcinomas from other poorly differentiated mammary epithelial tumors.

Lectin-Binding Patterns in the Spermatogenic Cells of the Shiba Goat Testis.

Lectin-binding patterns in the testis of the sexually mature goat were investigated by light and transmission electron microscopy. Dolichos biflorus agglutinin (DBA) and Griffonia simplicifolia agglutinin-I (GS-I) were negative in the seminiferous epithelium, but soybean (Glycine max) agglutinin (SBA), Griffonia simplicifolia agglutinin-II (GS-II) and peanut (Arachis hypogaea) agglutinin (PNA) were positive in the acrosomal vesicle of the Golgi-phase spermatids and in the acrosome of the cap-, acrosome- and maturation-phase spermatids. In addition, PNA was positive also in the plasma membrane and cytoplasm of the spermatogenic cells from the late pachytene spermatocytes to the maturation-phase spermatids. This finding indicates that glycoconjugates containing D-galactose residue appear at this period. Therefore, PNA may be a useful marker of developing spermatogenic cells. Electron microscopically, the positive reactions of SBA, GS-II and PNA were demonstrated in the central dense area (acrosomal granule) of the cap-phase spermatid acrosome, but not in the peripheral low-dense area. These results indicate that the lectin-bindings in the goat testis not only are common features, but also have patterns somewhat different from those in other mammals in such a way that GS-II can be detected in the acrosomal region of all phases of spermatids and that PNA is positive in the late spermatid acrosome.

Lectin histochemical characteristics of the canine female mammary gland

SUMMARY Twelve biotinylated lectins and an avidin-biotin-peroxidase method were used to detect and localize specific carbohydrate residues on formalin-fixed, paraffin-embedded female canine mammary gland sections. Histologic sections from 3 lactating and 7 nonlactating mixed-breed dogs (age 5.6 ± 0.35 years) were incubated with Arachis hypogea agglutinin (peanut agglutunin; pna), Concanavalia ensiformis agglutinin (conA), Dolichos biflorus agglutinin (dba), Glycine max agglutinin (sba), Griffonia simplicifolia agglutinin-I (gs-I), Lens culinaris agglutinin (lca), Lycopersicon esculentum agglutinin (lea), Phytolacca americana mitogen (pokeweed mitogen; pwm), Ricinus communis agglutinin-I and-II (rca-I and -II), Triticum vulgaris (wga), and Ulex europaeus agglutinin-I (uea-I). Each lectin had a specific binding pattern, except sba and dba. In nonlactating glands, pna, conA, lea, and uea-I stained duct cells in a linear-binding pattern, with a mean percentage of positive ducts per section of 28.7 (± 0.6), 65.7 (± 0.3), 100 (± 0), and 8.4 (± 0.2), respectively. Strong apical, lateral, basal, and cytoplasmic positivity on duct cells was seen after incubation of the sections with rca-I, rca-II, and wga in all ducts. In acinar cells, the binding pattern and the staining distribution of all the lectins studied were similar to those in duct cells. However, for pna, conA, and uea-I, the mean percentage of positive lobules per section was 33.7 (± 0.9), 62 (± 0.5), and 10.5 (± 0.2), respectively. In glands from lactating dogs, conA and uea-I did not stain. The cytoplasm of all myoepithelial cells was moderately stained with rca-I, rca-II, and wga. Endothelial cells stained with gs-I, pwm, rca-I, rca-II, wga, conA, and lca. The extracellular matrix, especially the periacinar and periduct regions, and the interstitial fibroblasts were positive for lca, rca-I, rca-II, and wga. Peripheral unmyelinated nerve fibers of the nipple were strongly positive for GS-I, pwm, rca-I, rca-II, and wga. Some of the lectins used (ie, pna, conA, uea-I, gs-I, pwm, and lea) appear to have selective staining of mammary gland structures that seems to be correlated with various physiologic functions. The contrasting binding pattern of lectins specific for the same sugar indicates a lack of knowledge of interactions between lectins and carbohydrate residues in tissue sections.

Difference in the ability of blood group-specific lectins and monoclonal antibodies to recognize the ABH antigens in human tissues

Twelve different kinds of blood group-specific lectins have been used along with monoclonal anti-A, -B and -H antibodies for detecting the corresponding antigens in selected human tissues. Although most of the lectins recognized the antigens in the tissue sections examined, they displayed marked differences in their recognition patterns in certain tissues. Helix asparsa agglutinin (HAA), Helix pomatia agglutinin (HPA) and monoclonal anti-A antibody recognized A antigens in the mucous cells of salivary glands from blood group A or AB nonsecretor as well as secretor individuals, whereas Dolichos biflorus agglutinin (DBA), Griffonia simplicifolia agglutinin-I (GSA-I), Sophora japonica agglutinin (SJA) and Vicia villosa agglutinin (VVA) did not bind to them from nonsecretors. A antigens in endothelial cells, lateral membrane of pancreatic acinar cells and small mucouslike cells of submandibular glands from some individuals were likewise recognized by HAA and HPA but not by other blood group A-specific lections. In contrast, both HAA and HPA did not recognize the A antigens in mucous cells of Brunner's glands while other A-specific lectins and monoclonal anti-A antibody reacted specifically with the antigens. Such a difference was not observed with lectins specific for blood group B. However, the B antigens in Brunner's glands were recognized by these lectins but not with monoclonal anti-B antibody. The difference in labelling ability was also noted among the blood group H-specific lectins and monoclonal anti-H antibody in endothelial cells of blood vessels. Ulex europaeus agglutinin-I reacted with these cells irrespective of ABO and the secretor status of the individuals, while Anguilla anguilla agglutinin and monoclonal anti-H antibody reacted only with those cells from blood group O individuals. No reaction was observed with Lotus tetragonolobus agglutinin in these tissue sites. These results suggest a great diversity of blood group antigens in different human tissues.

Cytochemistry for Lectins, Actin, Nucleotide Tetrazolium Reductases and Several Phosphatases in the Porcine Semitendinosus Muscle: Vascular Development in Young Pigs

An ontogeny study of the porcine semitendinosus muscle was conducted to meet two objectives: 1) to evaluate enzyme histochemistry, lectin cytochemistry and actin staining for usefulness as quantitative markers of muscle capillaries and 2) to describe the ontogeny of capillary density changes in developing porcine muscle. Muscle samples were obtained from three to eight crossbred pigs at each of the following ages: newborn, 1 to 2 d and 1, 2, 3, 4.5 and 24 wk. Cryostat sections were stained or reacted for alkaline and neutral phosphatase, dehydrogenase, actin, a panel of nine plant lectins (fluorescein isothiocyanate conjugated) and routine cytochemical tests for muscle fiber typing. Capillaries were quantitatively marked by reactions for dehydrogenase activity in the young pigs but not in the oldest animals. The lectins, soybean agglutinin, Bauhinia purpuria agglutinin, Ulex europeus agglutinin-I and Griffonia simplicifolia agglutinin-I (GS-I) quantitatively stained capillaries at all ages. Histological observations of thin sections of epon-embedded tissues served to validate the lectin and cytochemical capillary staining. Micrographs of sections stained with the lectin GS-I were used to count capillaries and muscle fibers so the capillary:fiber ratio (C/F) could be calculated. Deep (red) and superficial (white) aspects of muscle sections had different C/F at birth, 2, 4 and 24 wk. The deep aspects (DA) had higher C/F than superficial (SA) aspects (at all four ages), and there were age-dependent increases (P less than .001) in C/F of DA and SA. This study demonstrates the usefulness of lectin staining for determining C/F in porcine muscle.

Cytochemical localization of blood group substances in human salivary glands using lectin-gold complexes.

We investigated localization of blood group antigens and their related substances in human labial salivary and submandibular glands by application of a post-embedding cytochemical staining procedure using lectin- or glycoprotein-gold complexes. Surgical tissue was obtained from 10 patients. Blood group-specific lectins, such as Dolichos biflorus agglutinin or Helix pomatia agglutinin (group A-specific), Griffonia simplicifolia agglutinin-I B4 (group B-specific), and Ulex europaeus agglutinin I (group H-specific) could recognize A, B, and H antigens, respectively, only in mature secretory granules (mature SG), which were found preferentially in cells in the late phase of the maturation cycle. In immature secretory granules (immature SG), which were found in cells in the early or middle phase of the maturation cycle, no binding with these lectins was observed. The Golgi complexes and endoplasmic reticula also were not labeled with these lectins. In blood group O and B secretors, blood group antigens were uniformly distributed throughout all the mature SG examined. However, in blood group A secretors, the distribution was heterogeneous, i.e., in some granules only H antigen was demonstrated, whereas in others both A antigens and a small amount of H antigens were detected. Among the blood group-nonspecific lectins, wheat germ agglutinin (WGA) was found to bind more preferentially to immature SG than to mature SG. This was demonstrated irrespective of the blood group and secretor status of the tissue donor, except that in blood group A secretors WGA bound strongly to some mature SG which possessed A antigen. We discuss the significance of cellular and subcellular mosaic distribution of blood group antigens in connection with morphological differences of secretory granules and the maturation cycle of mucous cells.

Lectin histochemistry and ultrastructure of feline kidneys from six different storage diseases.

We have compared the pattern of lectin staining with the ultrastructural features of kidneys from normal cats and 19 cats with 6 different lysosomal storage diseases. The diseases studied include GM1 and GM2 gangliosidosis, mucopolysaccharidosis (MPS)-I and MPS-VI, sphingomyelin-lipidosis (i.e., Niemann-Pick disease) and mannosidosis. Ten different biotinylated lectins were used as histochemical probes for carbohydrate residues and avidin-biotin-peroxidase complex as visualant. Concanavalia ensiformis agglutinin (Con A) stained mesangial cells in all storage diseases but GM1, epithelial cells in sphingomyelin-lipidosis and mannosidosis, endothelial cells in GM1 and mannosidosis and Bowman's capsule cells in all but GM2. Griffonia simplicifolia agglutinin I (GS-I) stained the glomerular endothelium in all six diseases, but not in control kidneys. Ricinus communis agglutinin-I (RCA-I) stained the glomerular epithelium only in GM1 and MPS-I. Succinylated wheat germ agglutinin (SWGA) stained the glomerular endothelium and epithelium in mannosidosis, and the glomerular epithelium and Bowman's capsule in MPS-I. Ultrastructure studies demonstrated an accumulation of oligosaccharides in cases of mannosidosis and GM1 gangliosidosis, a mixture of oligosaccharides and lipids in MPS-I, MPS-VI and GM2 gangliosidosis and only lipid storage in sphingomyelin lipidosis. These studies show that morphologic and histochemical changes are manifested in some kidney cell types in lysosomal storage diseases, even though the enzyme deficiency occurs in all cell types. Furthermore, we show that the nature of the undegraded stored material is complex and that other factors, such as rate of membrane turn over, membrane composition, and cell function may influence the amount and nature of the "stored" material.

Histochemical demonstration of sugar residues by lectin and immunocytochemical techniques for blood group antigens in human colon.

Goblet cell mucin in 39 human colons was studied by methods specific for various sugar residues, including staining with three lectins, Dolichos biflorus agglutinin (DBA, specific for blood group A antigen), Griffonia simplicifolia agglutinin-I (GSA-I, B) and peanut agglutinin (PNA, T antigen), and immunostaining for A, B, H and T. Isoantigens A, B or H were found only in the right colon. GSA-I reactive goblet cells occurred in the right colon of both blood group A and B patients and possibly contained isoantigens. However DBA reactive cells were found in all cases. Prior neuraminidase digestion imparted anti-A, GSA-I and DBA reactivities to the cells lining the lower crypts in all cases. This pretreatment also imparted PNA and anti-T reactivities to goblet cells, only the latter reactivity being eliminated by galactose oxidase. Goblet cell mucin in transitional mucosa revealed decreased A and B, and increased H antigens. Enhanced galactose oxidase-Schiff (GOS) and anti-T reactivities were also noted. The present results revealed that some lectin reactions of goblet cells might be related to blood group antigens but others were not, and that different techniques for demonstrating reputedly the same sugar residues produced different results, indicating a need for proper evaluation of their specificity.

The glycoprotein-bound large carbohydrates from embryonal carcinoma cells carry receptors for several lectins recognizing N-acetylgalactosamine and galactose.

When various lectins were mixed with radioactively labeled embryoglycan (polylactosamine-type glycoprotein-bound carbohydrates from early embryonic cells) isolated from F9 embryonal carcinoma cells and the resulting complex was precipitated with ammonium sulfate, the glycan was found to react with the following lectins: Helix pomatia agglutinin (HPA), soybean agglutinin (SBA), Sophora japonica agglutinin (SJA), and Ricinus communis agglutinin-1 (RCA-1). Furthermore, affinity chromatography on lectin-agarose revealed that receptors for Griffonia simplicifolia agglutinin-I (GS-I) were also carried by the glycan. Together with the previous finding that the glycan carries receptors for Dolichos biflorus agglutinin (DBA) and peanut agglutinin (PNA), the present result established that the glycan has receptors for a variety of lectins recognizing N-acetylgalactosamine and/or galactose in teratocarcinoma cells. Intact molecules carrying GS-1 receptors and SJA receptors were isolated from F9 cells and teratocarcinoma OTT6050 and were shown to be high-molecular weight glycoproteins similar to DBA receptors isolated from the same sources.