Role Of Glycosaminoglycans Research Articles

A role of GAGs in ECM on morphogenesis of megakaryocytes.

The morphogenesis of megakaryocytes that results in the formation of cytoplasmic processes is thought to be the final maturation step before liberation of platelets. We studied the in vitro effects of glycosaminoglycans (GAGs) which are abundant in the bone marrow extracellular matrices, on the morphogenesis of murine megakaryocytes and compared them with those of thrombopoietic cytokines. Heparin, heparan sulfate, chondroitin-6 sulfate, and dermatan sulfate promoted the formation of megakaryocytic processes. Hyaluronic acid failed to support this phenomenon, suggesting that sulfated GAGs in extracellular matrices are involved in the morphogenesis of megakaryocytes. Sulfated GAGs began to act on megakaryocytes with a higher ploidy (16N-32N) from 6 to 24h after incubation, whereas neither rhIL-6 nor rhIL-11 affected this early phase. Our findings indicate that sulfated GAGs promote the morphogenesis of murine megakaryocytes and participate in thrombopoiesis in a different manner from that of cytokines such as rhIL-6 and rhIL-11.

Time and position-specific expression of glycosaminoglycans in rat molar cementum related to physiological tooth movement.

The role of glycosaminoglycans (GAGs) and proteoglycans during cementogenesis is not known. In this study, we have analysed the temporal and spacial expression of GAGs in the cellular cementum of 10-30 weeks old rats, immunohistochemically using monoclonal antibodies 2B6 and 3B3, specific for chondroitin 4-sulfate/dermatan sulfate and chondroitin 6-sulfate, respectively. Both 2B6- and 3B3-epitopes were expressed at similar position and time in the rat cellular cementum. Two types of cellular cementum were identified; GAG-positive and GAG-negative cementum. The former corresponded to the lightly stained and the latter to the darkly stained cementum in sections stained with haematoxylin and eosin. The GAG-positive cementum was seen at the distal side of dentine surface and appeared most thick at, middle of the apical half roots, whereas the other parts of the cementum were the GAG-negative. Distribution of GAG-positive cementum showed changes with age of animals. In 10-15 week old rats, the GAG-positive cementum occupied most of the cementum layer, covering a thin layer of the GAG-negative cementum. The cellular cementum of 20-30 week old rats consisted of three layers; GAG-negative, GAG-positive and GAG-negative cementum from dentine to cementum surface, reducing the GAG-positive area. Because our previous study has demonstrated that the lightly stained cementum is uncalcified, the present result suggests a correlation between calcification and contents of GAGs in the cellular cementum. Further, time- and position-specific expression of GAGs indicates their relation to the physiological tooth movement, which has been known in the rat molars.

Extracellular matrix in neuronal regeneration. Selective role of glycosaminoglycans

Extracellular matrix in neuronal regeneration. Selective role of glycosaminoglycans

High-molecular-weight proteins of nontypeable Haemophilus influenzae mediate bacterial adhesion to cellular proteoglycans

A family of high-molecular-weight (HMW) surface-exposed proteins of nontypeable Haemophilus influenzae (NT H. influenzae) mediated adherence of these organisms to human epithelium. To better understand the molecular basis for this adherence, the role of glycosaminoglycans (GAGs), substances commonly expressed on cell surfaces, was examined. Bacterial adherence to cells with specific deficiencies in GAG biosynthesis was measured. HMW protein-dependent bacterial adherence to normal cells was significantly greater than adherence to cells deficient in sulfated GAGs or to cells deficient in heparan sulfate but overexpressing chondroitin sulfate. Cells expressing undersulfated heparan sulfate exhibited intermediate levels of bacterial adherence. The addition of exogenous dextran sulfate or heparin inhibited over 70% of the adherence of NT H. influenzae to normal cells, whereas hyaluronic acid and chondroitin sulfate tested at the same concentration (100 micrograms/ml) inhibited bacterial adherence by less than 11%. Treatment of cells with heparinase significantly reduced bacterial adherence. Following electrophoretic separation, HMW proteins were shown to bind directly to radiolabeled heparin. These results indicate that HMW protein-dependent adherence of NT H. influenzae is mediated by cellular sulfated GAGs and that heparan sulfate may be the predominant GAG involved in this process. However, the decreased adherence of bacteria to cells expressing undersulfated heparan sulfate and the inhibition of bacterial adherence by the addition of exogenous dextran sulfate suggest that bacterial adhesion to mammalian cells is likely to be influenced by a variety of factors, including the degree of sulfation and the specificity of the carbohydrate moieties contained in the cellular proteoglycans.

Structure, function, and pathology of proteoglycans and glycosaminoglycans in the urinary tract.

The roles of glycosaminoglycans and proteoglycans in the physiology of the urinary tract are reviewed. The structures of proteoglycans and glycosaminoglycans are reviewed together with their role in control of epithelial differentiation through stromal-epithelial interactions and as modulators of cytokines. Heparan sulfate proteoglycans appear to be important in maintaining selectivity of the kidney tubular basement membrane, and the majority of the glycosaminoglycan found in the urine appears to come from the upper tract. Evidence suggesting that a dense layer of glycosaminoglycans on the urothelial surface is important to maintaining urothelial impermeability is reviewed and new data showing a high density of proteoglycans on the lumenal surface of the urothelium is presented. The role of this layer in maintaining antibacterial adherence and impermeability was discussed together with data suggesting that failure of this layer is an etiologic factor in interstitial cystitis. A model of the bladder surface is also presented to illustrate the role of proteoglycans and exogenous glycosaminoglycans in the defenses of normal bladder lumen and the failure of these defenses in the interstitial cystitis bladder.

Glycosaminoglycan Antibodies in Endocrine Ophthalmopathy

An increased accumulation of glycosaminoglycans (GAG) in the orbita has been reported in endocrine ophthalmopathy (EO). In this study we investigated whether antibodies directed against GAG are present in the sera of 52 EO patients and 47 healthy controls. Three out of 52 patients exhibited low titers of antinuclear antibodies and all patients were negative for antibodies against extractable nuclear antigens. Isotype IgG antibodies were detected by means of an ELISA using hyaluronic acid and dermatan sulfate as antigens. Values were expressed as optical density at 405 nm. In comparison to the control group (0.445, 0.364, 0.588; median, 25th, 75th percentile) significantly (p < 0.001) higher hyaluronic acid antibody levels were found in EO patients (0.626, 0.473, 0.801). Untreated patients had significantly higher hyaluronic acid antibody values compared to controls (0.770 vs 0.437, median, p < 0.005). Thyroid status or antithyroid treatment did not influence binding of hyaluronic acid antibodies. When screening for dermatan sulfate antibodies no such difference could be observed between patients and controls. Thus, the detection of GAG antibodies in EO patients emphasizes the important role of glycosaminoglycans in the pathogenesis of thyroid eye disease.

A role for heparan sulfate in androgen-induced deoxyribonucleic acid synthesis of mouse mammary carcinoma (Shionogi carcinoma 115)-derived SC-3 cells.

In response to androgen stimuli, SC-3 cells cloned from mouse mammary carcinoma (Shionogi carcinoma 115) secrete heparin-binding growth factor, which is able to bind to fibroblast growth factor (FGF) receptor, ultimately resulting in SC-3 cell proliferation. A role for glycosaminoglycan in the process of this androgen-induced autocrine loop was examined. When quiescent SC-3 cells were stimulated with testosterone, enhanced DNA synthesis was demonstrated even at the early phase (12-24 h) of stimulation. During this early period, autocrine growth-promoting activity was found to be associated with cell surface and extracellular matrix, but not to be present in the conditioned medium. This bound form of growth-promoting activity was able to be extracted with 2 M NaCl (pH 7.5) and absorbed onto a heparin-Sepharose affinity column, from which it was eluted at a concentration of 1.1-1.3 M NaCl. Extracted growth factor, whose activity was partially blocked by anti-basic FGF antibody, up-regulated the expression of FGF receptor-1 mRNA. These characteristics were similar to those of a soluble form of SC-3-derived growth factor previously reported from our laboratory. Androgen-induced enhancement of DNA synthesis was inhibited by simultaneous treatment of SC-3 cells with 10 mIU/ml heparitinase or 25 mM sodium chlorate (an inhibitor of phosphoadenosine sulfate synthesis). However, chlorate treatment did not affect the synthesis and distribution of androgen-induced growth factor, suggesting that the response of chlorate-treated cells to this growth factor was impaired. These results indicate that heparan sulfate has important roles in concentrating androgen-induced heparin-binding growth factor on or very close to the cell surface and in potentiating its bioactivity.

P9. Control of chondrocyte maturation: role of glycosaminoglycans, c-myc and TGFβ in the development of avian tibial dyschondroplasia

P9. Control of chondrocyte maturation: role of glycosaminoglycans, c-myc and TGFβ in the development of avian tibial dyschondroplasia

Role of glycosaminoglycans in the regulation of T cell proliferation induced by thymic stroma-derived T cell growth factor.

The present study investigates the regulatory effects of glycosaminoglycans such as heparin and heparan sulfate on T cell proliferation induced by thymic stromal cell monolayer or its derived T cell growth factor (TCGF). A thymic stromal cell clone (MRL104.8a) supported the growth of Ag-specific, IL-2-dependent Th cell clone (9-16) in the absence of Ag and IL-2 by producing a unique TCGF designated as thymic stroma-derived T cell growth factor (TSTGF). The addition of heparin to cultures in which the growth of 9-16 Th cells was otherwise stimulated by the MRL104.8a monolayer or a semipurified sample of the TSTGF resulted in heparin dose-dependent inhibition of 9-16 Th proliferation. The dose of heparin required for inducing 50% reduction of TSTGF-induced proliferation of Th at a given cell number was found to be proportional to the magnitude of the TSTGF added to cultures, suggesting that heparin exerted its inhibitory effect by binding to the TSTGF rather than by acting on Th cells. A similar growth-inhibiting effect of heparin was observed in IL-7-dependent proliferation of pre-B cell line or Th, but not in IL-2-dependent T cell proliferation or IL-3-dependent myeloid cell proliferation. A strong affinity of TSTGF and IL-7 for heparin was confirmed by the fact that both TSTGF and IL-7 adhered to columns of heparin-agarose and were eluted by salt. When various glycosaminoglycans were tested for the heparin-like Th growth-regulatory capacity, heparan sulfate exhibited Th growth-inhibiting ability comparable to that observed for heparin. These results indicate that the activity of thymic and/or bone marrow stroma-derived lymphocyte growth factor (TSTGF/IL-7) but not of Th-producing TCGF (IL-2) is negatively regulated by heparin or heparan sulfate, which would represent major glycosaminoglycans in the extra-cellular matrix of stromal cells.

Visualization of glycosaminoglycans in rat incisor extracellular matrix using a hyaluronidase-gold complex

The enzyme-gold technique was used on dental tissues. Hyaluronidase was complexed with gold, and ultrathin sections of rat incisors were incubated with the hyaluronidase-gold complex to localize chondroitin-sulphate and hyaluronic acid at the ultrastructural level. The hyaluronidase-gold complex was found in predentine and dentine, especially at the mineralization front, in interglobular spaces and around dentinal canaliculi. The very young enamel was labelled, but not the later stages of formation. This method allowed a very precise localization of hyaluronic acid and/or chondroitin sulphate in rat incisors extracellular matrices. These findings support the important role of glycosaminoglycans in dentine mineralization.

Chondroitin sulfates in developing mouse tooth germs

Abstract The role of glycosaminoglycans and proteoglyans during ontogenesis is not known. The developing tooth offers a potentially important model for studies of structure-function relationships. In this study, we have analysed the temporal and spatial expression of chondroitins of differing sulfation patterns in embryonic molars and incisors. For this purpose, we have used monoclonal antibodies (Mabs) specific for unsulfated, 4-sulfated, and 6-sulfated forms of chondroitin in conjunction with indirect immuno-fluorescence or immunoperoxidase labeling. Unsulfated chondroitin was not detected in embryonic teeth. Chondroitin 4- and chondroitin 6-sulfates were present in the stellate reticulum but otherwise they were confined to the dental mesenchyme. The 3B3 and MC21C-epitope, which are markers of 6-sulfated chondroitin, were uniformly distributed in the dental mesenchyme during the bud stage; they disappeared from the dental papilla of the cusps and of the anterior region of the incisor as development proceeded. These epitopes were absent from the basement membrane and from the predentin. In the odontoblastic cell lineage, the 3B3 and MC21C-epitopes were detected only between preodontoblasts at an early stage of differentiation. The monoclonal antibody 2B6 served as a probe to localize chondroitin 4-sulfate. This glycosaminoglycan was detected as early as the dental lamina stage but its expression was restricted to the basement membrane of the teeth until the late bell stage. After the onset of cusp formation, strong staining was also observed over the occlusal region of the dental papilla while the cervical region of the dental papilla remained 256-negative. Incisors at the bell stage exhibited a decreasing gradient of immunostaining by 2B6 from their anterior region to their posterior end. The extracellular matrix surrounding preodontoblasts reacted with 2B6 and the predentin, produced by the odontoblasts, was also intensely labeled with this antibody. Comparison between immunostaining with 3B3 and 2B6, on consecutive sections revealed a mutually exclusive pattern of distribution of the corresponding epitopes during odontogenesis. Furthermore, in the continuously growing incisor, a striking positive correlation was found between the immunostaining patterns produced by 3B3 and MC21C and the mitotic indices along the anterior-posterior axis of the tooth. Hence, sulfation of chondroitin seems developmentally regulated. We postulate that changes in the sulfation pattern of chondroitin might play a role in ontogenesis by locally altering the functional properties of the extracellular matrix.

Neural tube defects. Some remarks on the possible role of glycosaminoglycans in the genesis of the dysraphic state, the anomaly in the configuration of the posterior cranial fossa, and hydrocephalus.

Recent developments in the field of experimentally induced neural tube defects (NTD) indicate that specific substances, namely the glycosaminoglycans (GAGs) may play a role in the genesis of spinal malformations. The authors report the results obtained by evaluating the GAGs in rat fetuses with NTD, secondary to the administration of Trypan Blue during pregnancy. A characteristic decrease in GAGs formation in the spinal and cranial structures as well as in the subependymal regions of the brain was found in the malformed fetuses. The authors hypothesize that this anomaly in GAGs formation is responsible for both the NTD and the associated malformations, namely hydrocephalus and hypoplasia of the posterior cranial fossa.

Role of glycosaminoglycan and fibronectin in endothelial cell growth

This study was undertaken to clarify the biological significance of fibronectin and GAG synthesized in human endothelium. The synthesis of GAG and fibronectin was proved morphologically and biochemically in endothelial cells. The conditioned media of fibroblasts and smooth muscle cells have promoted endothelial cell proliferation and production of extracellular matrix in the endothelium. These results suggest that the extracellular matrix which is composed of fibronectin and GAG may play an important role in endothelial cell regeneration.

Role of glycosaminoglycans in calcium metabolism of human platelets

Changes in intracellular Ca 2+, [Ca 2+] i, were measured in control and chondroitin ABC lyase-pretreated platelets. [Ca 2+] i was measured with the fluorescent calcium probe Quin2. Chondroitin ABC lyase removed chondroitin 4-sulfate from the platelet surface without inducing shape change or release of serotonin. Compared to similarly prepared controls, enzyme treated platelets showed an increase of [Ca 2+] i in response to stimulation by various agonists at high (1 mM) extracellular Ca 2+ concentration. At low Ca 2+ in the medium (1 mM EGTA), such platelets responded to agonists with a decreased rise in [Ca 2+] i compared to the controls. These studies indicate that selective removal of glycosaminoglycans may sensitize platelets to the action of platelet aggregating agents. In addition, glycosaminoglycans may have a calcium storage function.

The role of glycosaminoglycans in anuran pigment cell migration

The migratory pathways of neural crest-derived pigment cells were examined in two anurans, Xenopus laevis and Discoglossus pictus, and correlated with the distribution of glycosaminoglycans (GAG) in the extracellular matrix (ECM) of these pathways. In Xenopus, melanophores in the trunk reach the dermis by initially migrating ventrally, between the neural tube and somites, and then by migrating through the somites to reach the subectodermal space. In Discoglossus, melanophores, iridophores, and xanthophores migrate laterally over the dorsal margin of the somites to reach the dermis. GAG was identified in the light microscope using alcian blue staining and in the electron microscope using ruthenium red staining. The ECM at the dorsal entrance to the lateral pathway in Xenopus and in young Discoglossus (at a stage prior to invasion by pigment cells) is filled with 25-50 nm chondroitin sulphate proteoglycan aggregates. When this ECM in Xenopus is digested in vivo with chondroitinase ABC, melanophores enter the lateral pathway. In older Discoglossus embryos, the migration of pigment cells into the lateral pathway is correlated with increases in the space between the ectoderm and somites and in the number of hyaluronate microfibrils. These observations suggest that chondroitin sulphate proteoglycan in the subectodermal ECM restricts the migration of pigment cells into the lateral pathway by limiting the amount of space for migration and possibly by acting as a less adhesive migratory substratum than the ventral pathway, and that in Discoglossus hyaluronate opens spaces permitting the migration of pigment cells directly over the dorsal margin of the somites.

The lung parenchyma--a dynamic matrix. J. Burns Amberson lecture.

The matrix of the lung parenchyma is composed predominantly of collagen, elastin, glycosaminoglycans, and fibronectin. This presentation focuses on the newer insights into injury and repair of lung elastin and glycosaminoglycans. Elastin and glycosaminoglycans respond in characteristic patterns to destructive and proliferative forms of lung injury. Elastin degradation induced by elastases induces a prompt and marked response by elastin resynthesis in situ. The signal for the stimulus remains unknown and deserves to be understood. The measurement of elastin peptides in plasma, urine, or bronchoalveolar lavage fluid can provide an objective index of elastin degradation in disease. Elastin depletion may be augmented by factors that interfere with elastin resynthesis such as exposure to tobacco smoke. Glycosaminoglycans participate in similar fashion in many different forms of lung injury as an early response to injury, which may then determine the subsequent cellular processes of repair. Studies of injury and repair in organ systems, other than the lung, suggest a general role for glycosaminoglycans in interstitial tissue and repair. The mediators that alter matrix chemical structure and function could provide powerful instruments for controlling the process of tissue injury and repair in the lung.

A role for glycosaminoglycans in cellular adhesion of relevance to the cancer state

Conference Article| April 01 1985 A role for glycosaminoglycans in cellular adhesion of relevance to the cancer state DAVID GRANT; DAVID GRANT 1Department of Biochemistry, University of Aberdeen, Marischal College, Aberdeen AB9 1AS, Scotland, U.K. Search for other works by this author on: This Site PubMed Google Scholar WILLIAM F. LONG; WILLIAM F. LONG 1Department of Biochemistry, University of Aberdeen, Marischal College, Aberdeen AB9 1AS, Scotland, U.K. Search for other works by this author on: This Site PubMed Google Scholar FRANK B. WILLIAMSON FRANK B. WILLIAMSON 1Department of Biochemistry, University of Aberdeen, Marischal College, Aberdeen AB9 1AS, Scotland, U.K. Search for other works by this author on: This Site PubMed Google Scholar Biochem Soc Trans (1985) 13 (2): 389. https://doi.org/10.1042/bst0130389 Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Facebook Twitter LinkedIn MailTo Cite Icon Cite Get Permissions Citation DAVID GRANT, WILLIAM F. LONG, FRANK B. WILLIAMSON; A role for glycosaminoglycans in cellular adhesion of relevance to the cancer state. Biochem Soc Trans 1 April 1985; 13 (2): 389. doi: https://doi.org/10.1042/bst0130389 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsBiochemical Society Transactions Search Advanced Search Keywords: NIR spectroscopy, near-infrared spectroscopy This content is only available as a PDF. © 1985 Biochemical Society1985 Article PDF first page preview Close Modal You do not currently have access to this content.

Role of glycosaminoglycans and collagen in the development of a fibronectin-rich extracellular matrix in cultured embryonic corneal epithelial cells.

Corneal epithelial cells from 15-day chick embryos produce a fibronectin-rich extracellular matrix when cultured on glass, plastic and fibronectin-coated substrata. Cell culture in the presence of Streptomyces hyaluronidase or chondroitinase ABC resulted in considerable reduction of the matrix; collagenase had a lesser effect but nevertheless also reduced the matrix. In all enzyme treatments the cells attached and spread to form characteristic epithelial cell islands, but the marginal cells of these islands showed a marked reduction in the number of lamellipodia and focal contacts. Also, the immunofluorescent staining pattern for fibronectin was considerably reduced. Control cells cultured on a fibronectin-coated surface were able to reorganize the fibronectin into fibrils, whereas cells cultured in enzymes showed little or no ability to do so. The cellular reorganization of fibronectin could also be inhibited by the addition of L-azetidine-2-carboxylic acid (LACA), an inhibitor of collagen secretion. Cells plated out in the presence of LACA spread much better on collagen substrata than on plastic, glass or fibronectin. However, in all cases very little fibronectin matrix was detectable in the epithelial islands. The results suggest that components of the extracellular matrix (ECM) such as collagen, hyaluronic acid and chondroitin sulphates are not essential for the initial attachment and spreading of corneal epithelial cells in culture, but are important in the development of the ECM, and in maintaining a flattened morphology and spreading behaviour. It is suggested that fibronectin plays an important role in these interactions and that the ability of cells to organize fibronectin into fibrils is dependent on the presence of other ECM components such as glycosaminoglycans and collagen.

Type, location and role of glycosaminoglycans in cloned differentiated chick retinal pigmented epithelium

In clonal culture, colonies of 3–4 week old chick retinal pigmented epithelial cells exhibit Alcian Blue positive extracellular matrix (ECM) material on the surface of the cells. Alcian blue positive ECM is located between undifferentiated cells at the edges of the disc-shaped colonies and beneath the differentiated cells in the colony center. The latter material is associated with the basement membrane. The staining properties suggest that glycosaminoglycans (GAG) are present in these regions. Extraction of GAG from homogenates of colonies, followed by electrophoresis on cellulose acetate strips, results in three bands with mobilities similar to those of hyaluronic acid, heparan sulfate, and chondroitin sulfate, respectively. All three bands label with [ 3H]glucosamine, and the last two also label with [ 35S]sulfate. The composition appeared to differ when colonies were grown in different media. Digestion of the GAG preparations with various enzymes suggests that bands II and III represent heparan sulfate and chondroitin sulfate, respectively, in colonies grown in Ham's F10g medium. The composition of band I is as yet undetermined. In minimal Eagle's medium (MEM), bands I and III consisted of hyaluronic acid and chondroitin sulfate, respectively, while band II had properties suggestive of a copolymer of heparan sulfate and an unidentified GAG. Cells release only one [ 3H]glucosamine-labelled GAG into the medium. This material has a mobility similar to hyaluronic acid and is digested by Streptomyces hyaluronidase, suggesting that it is hyaluronic acid. Staining with Alcian Blue at different pH suggests that it may represent the material associated with the upper surface of the cells. Some of the ECM located between the undifferentiated cells and associated with the basement membrane in the differentiated regions of the colonies stains with Alcian Blue at pH 1.0 and 0.2 suggesting that it may contain GAGs found in bands I and II. Colonies treated with medium containing 6-diazo-5-oxo-L-norleucine (DON), an inhibitor of GAG synthesis, for 48 hr showed a reduced Alcian Blue staining of the ECM in the undifferentiated regions. After 72 hr of treatment with DON, the undifferentiated cells had detached from the plate, whereas the differentiated cells remained intact. The results suggest that the GAG may be involved in cellular adhesion, particularly of the undifferentiated cells.

A role for glycosaminoglycans in the development of collagen fibrils

Extensive data on the glycosaminoglycan (GAG) composition and the collagen fibril diameter distribution have been collected for a diverse range of connective tissues. It is shown that tissues with the smallest diameter collagen fibrils (mass-average diameter < 60 nm) have high concentrations of hyaluronic acid and that tissues with the largest diameter collagen fibrils (mass-average diameter ∼200 nm) have high concentrations of dermatan sulphate. It is suggested that the lateral growth of fibrils beyond a diameter of about 60 nm is inhibited by the presence of an excess of hyaluronic acid but that this inhibitory effect may be removed by an increasing concentration of chondroitin sulphate and/or dermatan sulphate. It is also postulated that high concentrations of chondroitin sulphate will inhibit fibril growth beyond a mass-average diameter of ∼150 nm. Such an inhibition may in turn be removed by an increasing concentration of dermatan sulphate such that it becomes the dominant GAG present in the tissue.