Density-dependent expression of ganglioside GM3 by human skin fibroblasts in an all-or-none fashion, as a possible modulator of cell growth in vitro

Rösner, Harald; Greis, Ch.; Rodemann, H. Peter

doi:10.1016/0014-4827(90)90180-i

Cited by 29 publications

(11 citation statements)

References 45 publications

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“…2) indicates a low number of Gb3Cer negative cells in the subconfluent culture. The same phenomenon, i.e., a non-homogeneous GSL-expression in pre-confluent cell cultures, has been reported for the immunochemical detection of GM3 in human fibroblasts by Rösner et al [57]. Thus, a cell density-and/or cell cycle-dependent expression could explain the observed faint heterogeneity of Stx1-receptor expression and cytotoxicity response in EA.hy 926 cells.…”

Section: Discussionsupporting

confidence: 79%

Glycosphingolipids in vascular endothelial cells: relationship of heterogeneity in Gb3Cer/CD77 receptor expression with differential Shiga toxin 1 cytotoxicity

et al. 2008

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Shiga toxin (Stx) 1 binds to the glycosphingolipid (GSL) globotriaosylceramide (Gb3Cer/CD77) and injures human endothelial cells. In order to gain insight into Stx1-induced cellular impairment, we analysed in detail the molecular heterogeneity of Stx1 receptors in two endothelial cell lines differing in their Stx1-sensitivity. We observed a moderate sensitivity to Stx1 of human brain microvascular endothelial cells (HBMECs, CD(50) > 200 ng/ml), but a considerably higher mortality rate in cultures of EA.hy 926 cells, a cell line derived from human umbilical vein endothelial cells (CD(50) of 0.2 ng/ml). Immunofluorescence microscopy demonstrated the presence of Gb3Cer in both cell lines, but showed an enhanced content of Gb3Cer in EA.hy 926 cells. Solid phase overlay binding assays of isolated GSLs combined with nanoelectrospray ionization quadrupole time-of-flight mass spectrometry demonstrated a balanced proportion of Gb3Cer and globotetraosylceramide (Gb4Cer) in HBMECs, but an increase of Gb3Cer and absence of Gb4Cer in EA.hy 926 cells. Gb3Cer species with C24:1/C24:0 fatty acids were found to dominate over those with C16:0 fatty acids in EA.hy 926 cells, but were similarly distributed in HBMECs. Reverse transcriptase polymerase chain reaction indicated the concomitant presence of Gb3Cer and Gb4Cer synthases in HBMECs, whereas EA.hy 926 cells expressed Gb3Cer synthase, but completely lacked Gb4Cer synthase. This deficiency, resulting in the accumulation of Gb3Cer in EA.hy 926 cells, represents the most prominent molecular reason that underlies the different Stx1 sensitivities of HBMECs and EA.hy 926 endothelial cells.

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Section: Discussionsupporting

confidence: 79%

Glycosphingolipids in vascular endothelial cells: relationship of heterogeneity in Gb3Cer/CD77 receptor expression with differential Shiga toxin 1 cytotoxicity

et al. 2008

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“…The role of a physical mechanism to inhibit cell division is a novel finding that has not been previously reported and may contribute to the well-known molec- The mechanism of contact inhibition has been previously attributed to an interaction of plasma membrane glycoproteins such as gangliosides with membrane receptor protein tyrosine phosphatases on an adjacent cell (25). Plasma membrane gangliosides are also known to be lost in transformed cancer cells (29). In addition to activation of receptor protein tyrosine phosphatases, cell-cell contact is thought to activate a protein called contactinhibin, which in turn inhibits mitogenic pathways (25,37).…”

Section: Discussionmentioning

confidence: 99%

Inhibition of fibroblast proliferation in cardiac myocyte cultures by surface microtopography

Boateng¹,

Hartman²,

Ahluwalia³

et al. 2003

American Journal of Physiology-Cell Physiology

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Cardiac myocyte cultures usually require pharmacological intervention to prevent overproliferation of contaminating nonmyocytes. Our aim is to prevent excessive fibroblast cell proliferation without the use of cytostatins. We have produced a silicone surface with 10-microm vertical projections that we term "pegs," to which over 80% of rat neonatal cardiac fibroblasts attach within 48 h after plating. There was a 50% decrease in cell proliferation by 5 days of culture compared with flat membranes (P < 0.001) and a concomitant 60% decrease (P < 0.01) in cyclin D1 protein levels, suggesting a G1/S1 cell cycle arrest due to microtopography. Inhibition of Rho kinase with 5 or 20 microM Y-27632 reduced attachment of fibroblasts to the pegs by over 50% (P < 0.001), suggesting that this signaling pathway plays an important role in the process. Using mobile and immobile 10-microm polystyrene spheres, we show that reactive forces are important for inhibiting fibroblast cell proliferation, because mobile spheres failed to reduce cell proliferation. In primary myocyte cultures, pegs also inhibit fibroblast proliferation in the absence of cytostatins. The ratio of aminopropeptide of collagen protein from fibroblasts to myosin from myocytes was significantly reduced in cultures from pegged surfaces (P < 0.01), suggesting an increase in the proportion of myocytes on the pegged surfaces. Connexin43 protein expression was also increased, suggesting improved myocyte-myocyte interaction in the presence of pegs. We conclude that this microtextured culture system is useful for preventing proliferation of fibroblasts in myocyte cultures and may ultimately be useful for tissue engineering applications in vivo.

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“…In addition, anti-Gg 4 Cer and anti-Gb 4 Cer antibodies did not react with B-cell-dependent zones in the secondary lymphoid organs, whereas they bound to B-cells in flow cytometry. A possible explanations for such discrepancy is a change in the surface density and composition of membrane GSLs and gangliosides during preparation of single-cell suspensions, which may affect recognition by anti-GSL antibodies (Nores et al 1987;Rösner et al 1990;Lloyd et al 1992;Hildebrandt 1996;Tatewaki et al 1997 ). Another explanation could lie in the influence of the fixative on the reliability of immunohistochemical studies (Prasadarao et al 1990;Schwarz and Futerman 1997).…”

Section: Discussionmentioning

confidence: 99%

Immunohistological and Flow Cytometric Analysis of Glycosphingolipid Expression in Mouse Lymphoid Tissues

Kovačić

Müthing

Marušić

2000

J Histochem Cytochem.

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Expression of neutral glycosphingolipids (GSLs) and gangliosides in normal lymphoid tissues and cells has been studied mostly by biochemical and immunochemical analysis of lipid extracts separated by thin-layer chromatography. GSLs and gangliosides involved in the GM1b biosynthetic pathway were assigned to T-lymphocytes, whereas B-cell gangliosides and GSLs have been poorly characterized in former publications. We used specific polyclonal antibodies in immunohistochemistry and flow cytometry to analyze the distribution of globotriaosylceramide (Gb3Cer), globoside (Gb4Cer), gangliotriaosylceramide (Gg3Cer), gangliotetraosylceramide (Gg4Cer), and gangliosides GM3 and GalNAc-GM1b in the mouse thymus, spleen, and lymph node. Immature thymocytes expressed epitopes recognized by all antibodies, except for anti-Gb4Cer. Mature thymocytes bound only antibodies to GalNAc-GM1b, Gg4Cer, and Gb4Cer. In secondary lymphoid organs, antibodies to globo-series GSLs bound to vascular spaces of secondary lymphoid organs, whereas the ganglio-series GSL antibodies recognized lymphocyte-containing regions. In a Western blotting analysis, only Gal-NAc-GM1b antibody recognized a specific protein band in all three organs. Flow cytometric analysis of spleen and lymph node cells revealed that B-cells carried epitopes recognized by all antibodies, whereas the T-cell GSL repertoire was mostly oriented to ganglio-series-neutral GSLs and GM1b-type gangliosides. The results of immunohistochemistry and flow cytometry were not always identical, possibly because of crossreactivity to glycoprotein-linked oligosaccharides and/or differences between cell surface carbohydrate profiles of isolated cells and cells in a tissue environment.

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Density-dependent expression of ganglioside GM3 by human skin fibroblasts in an all-or-none fashion, as a possible modulator of cell growth in vitro

Cited by 29 publications

References 45 publications

Glycosphingolipids in vascular endothelial cells: relationship of heterogeneity in Gb3Cer/CD77 receptor expression with differential Shiga toxin 1 cytotoxicity

Glycosphingolipids in vascular endothelial cells: relationship of heterogeneity in Gb3Cer/CD77 receptor expression with differential Shiga toxin 1 cytotoxicity

Inhibition of fibroblast proliferation in cardiac myocyte cultures by surface microtopography

Immunohistological and Flow Cytometric Analysis of Glycosphingolipid Expression in Mouse Lymphoid Tissues

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