scholarly journals Relationship of transformation, cell density, and growth control to the cellular distribution of newly synthesized glycosaminoglycan.

1976 ◽  
Vol 71 (1) ◽  
pp. 280-294 ◽  
Author(s):  
R H Cohn ◽  
J J Cassiman ◽  
M R Bernfield
Keyword(s):  
Hyaluronic Acid ◽  
Cell Surface ◽  
Heparan Sulfate ◽  
Chondroitin Sulfate ◽  
Cell Density ◽  
Growth Control ◽  
Cellular Distribution ◽  
High Cell ◽  
Density Dependent ◽  
Relationship Of

Mouse 3T3 cells and their Simian Virus 40-transformed derivatives (3T3SV) were used to assess the relationship of transfromation, cell density, and growth control to the cellular distribution of newly synthesized glycosaminoglycan (GAG). Glucosamine- and galactosamine-containing GAG were labeled equivalently by [3H=A1-glucose regardless of culture type, allowing incorporation into the various GAG to be compared under all conditions studied. Three components of each culture type were examined: the cells, which contain the bulk of newly synthesized GAG and are enriched in chondroitin sulfate and heparan sulfate; cell surface materials released by trypsin, which contain predominantly hyaluronic acid; and the media , which contain predominantly hyaluronic acid and undersulfated chondroitin sulfate. Increased cell density and viral transformation reduce incorporation into GAG relative to the incorporation into other polysaccharides. Transformation, however, does not substantially alter the type or distribution of newly synthesized GAG; the relative amounts and cellular distributions were very similar in 3T3 and 3T3SV cultures growing at similar rates at low densities. On the other hand, increased cell density as well as density-dependent growth inhibition modified the type and distribution of newly synthesized GAG. At high cell densities both cell types showed reduced incorporation into hyaluronate and an increase in cellular GAG due to enhanced labeling of chondroitin sulfate and heparan sulfate. These changes were more marked in confluent 3T3 cultures which also differed in showing substantially more GAG label in the medium and in chondroitin-6-sulfate and heparan sulfate at the cell surface. Since cell density and possibly density-dependent inhibition of growth but not viral transformation are major factors controlling the cellular distribution and type of newly synthesized GAG, differences due to GAG's in the culture behavior of normal and transformed cells may occur only at high cell density. The density-induced GAG alterations most likely involved are increased condroitin-6-sulfate and heparan sulfate and decreased hyaluronic acid at the cell surface.

Effect of 12-O-tetradecanoylphorbol-13-acetate on cell-surface glycosaminoglycans in human diploid fibroblasts

1984 ◽  
Vol 62 (12) ◽  
pp. 1354-1357
Author(s):  
Bruce R. Wolff ◽  
Bernard R. Glick ◽  
Niels C. Bols
Keyword(s):  
Hyaluronic Acid ◽  
Cell Surface ◽  
Heparan Sulfate ◽  
Chondroitin Sulfate ◽  
Human Fibroblasts ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
The Other ◽  
Human Skin Fibroblasts ◽  
Human Diploid Fibroblasts

The effect of 12-O-tetradecanoylphorbol-13-acetate (TPA) on the loss of glycosaminoglycans (GAGs) from the pericellular compartment of human skin fibroblasts was studied. GAGs from this compartment were analyzed by anion-exchange chromatography after cultures had been labelled with both [3H]glucosamine and [35S]sulfate and then chased in either the presence or absence of TPA. In both control and TPA cultures radioactivity was found in glycopeptides, hyaluronic acid, heparan sulfate (HS), and chondroitin sulfate. The amount of radioactivity that was found in HS was reduced in the TPA cultures, whereas the amounts in the other GAGs were essentially unchanged when control and treated cultures were compared. These results suggest that TPA stimulates the loss of HS from the surface of human fibroblasts.

scholarly journals Cell Density-Dependent Fibroblast Growth Factor-2 Signaling Regulates Syndecan-4 Expression in Cultured Vascular Endothelial Cells

2020 ◽  
Vol 21 (10) ◽  
pp. 3698 ◽  
Author(s):  
Takato Hara ◽  
Shiori Yabushita ◽  
Chika Yamamoto ◽  
Toshiyuki Kaji
Keyword(s):  
Endothelial Cells ◽  
Growth Factor ◽  
Heparan Sulfate ◽  
Fibroblast Growth Factor ◽  
Cell Density ◽  
Vascular Endothelial Cells ◽  
Fibroblast Growth Factor 2 ◽  
Vascular Endothelial ◽  
Fibroblast Growth ◽  
Density Dependent

Syndecan-4 is a member of the syndecan family of transmembrane heparan sulfate proteoglycans, and is involved in cell protection, proliferation, and the blood coagulation-fibrinolytic system in vascular endothelial cells. Heparan sulfate chains enable fibroblast growth factor-2 (FGF-2) to form a complex with its receptor and to transduce the cell growth signal. In the present study, bovine aortic endothelial cells were cultured, and the intracellular signal pathways that mediate the regulation of syndecan-4 expression in dense and sparse cultures by FGF-2 were analyzed. We demonstrated the cell density-dependent differential regulation of syndecan-4 expression. Specifically, we found that FGF-2 upregulated the synthesis of syndecan-4 in vascular endothelial cells via the MEK1/2-ERK1/2 pathway in dense cell cultures, with only a transcriptional induction of syndecan-4 at a low cell density via the Akt pathway. This study highlights a critical mechanism underlying the regulation of endothelial cell functions by proteoglycans.

scholarly journals Codistribution of heparan sulfate proteoglycan, laminin, and fibronectin in the extracellular matrix of normal rat kidney cells and their coordinate absence in transformed cells.

1982 ◽  
Vol 94 (1) ◽  
pp. 28-35 ◽  
Author(s):  
E G Hayman ◽  
A Oldberg ◽  
G R Martin ◽  
E Ruoslahti
Keyword(s):  
Cell Surface ◽  
Heparan Sulfate ◽  
Chondroitin Sulfate ◽  
Rat Kidney ◽  
Heparan Sulfate Proteoglycan ◽  
Chondroitin Sulfate Proteoglycan ◽  
Transformed Cells ◽  
Kidney Cells ◽  
Sulfate Proteoglycan ◽  
Normal Rat

We used antibodies raised against both a heparan sulfate proteoglycan purified from a mouse sarcoma and a chondroitin sulfate proteoglycan purified from a rat yolk sac carcinoma to study the appearance and distribution of proteoglycans in cultured cells. Normal rat kidney cells displayed a fibrillar network of immunoreactive material at the cell surface when stained with antibodies to heparan sulfate proteoglycan, while virally transformed rat kidney cells lacked such a surface network. Antibodies to chondroitin sulfate proteoglycan revealed a punctate pattern on the surface of both cell types. The distribution of these two proteoglycans was compared to that of fibronectin by double-labeling immunofluorescent staining. The heparan sulfate proteoglycan was found to codistribute with fibronectin, and fibronectin and laminin gave coincidental stainings. The distribution of chondroitin sulfate proteoglycan was not coincidental with that of fibronectin. Distinct fibers containing fibronectin but lacking chondroitin sulfate proteoglycan were observed. When the transformed cells were cultured in the presence of sodium butyrate, their morphology changed, and fibronectin, laminin, and heparan sulfate proteoglycan appeared at the cell surface in a pattern resembling that of normal cells. These results suggest that fibronectin, laminin, and heparan sulfate proteoglycan may be complexed at the cell surface. The proteoglycan may play a central role in assembly of such complexes since heparan sulfate has been shown to interact with both fibronectin and laminin.

scholarly journals Regulation of haemopoiesis in long-term bone marrow cultures. IV. Glycosaminoglycan synthesis and the stimulation of haemopoiesis by beta-D-xylosides.

1983 ◽  
Vol 96 (2) ◽  
pp. 510-514 ◽  
Author(s):  
E Spooncer ◽  
J T Gallagher ◽  
F Krizsa ◽  
T M Dexter
Keyword(s):  
Bone Marrow ◽  
Cell Surface ◽  
Heparan Sulfate ◽  
Chondroitin Sulfate ◽  
Glycosaminoglycan Synthesis ◽  
Fivefold Increase ◽  
Bone Marrow Cultures ◽  
Marrow Cultures ◽  
Stimulation Of

Sulfated glycosaminoglycans (GAGs) are distributed in consistent and distinctive patterns between the cell surface and the growth medium of haemopoietically active long-term bone marrow cultures. Heparan sulfate is the main cell surface component and chondroitin sulfate is the major sulfated species in the medium. When the cultures are supplemented with beta-D-xylosides a significant increase in chondroitin sulfate synthesis is observed but no stimulation of heparan sulfate synthesis occurs. The chondroitin sulfate accumulates in the culture medium in beta-D-xyloside-treated cultures but the composition of sulfated GAGs in cell-surface derived material is unaffected. beta-D-xylosides also stimulate the production of haemopoietic cells without any apparent alteration in the adherent stromal cells of the marrow cultures. Equivalent increases are obtained in cells at all stages of development so that a fivefold increase in pluripotent stem cells (CFU-S) is matched by fivefold increase in the granulocyte-macrophage progenitors (GM-CFC) and in mature granulocytes. The stimulation persists for many weeks in beta-D-xyloside-treated cultures. These results indicate that the sulfated GAGs may play an important role in the regulation of haemopoiesis.

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