Murine Neuroblastoma Cells Express Ganglioside Binding Sites on Their Cell Surface

Abstract

The ability of S20Y cholinergic, and N115 adrenergic, murine neuroblastoma cells to adhere to immobilized gangliosides was studied. Viable S20Y cells adhered more strongly to GM1-coated plastic wells than to those coated with GM2, GD1a, or GT1b. The oligosaccharide portion of GM1 inhibited adherence of S20Y cells to GM1-coated wells, indicating that the carbohydrate moiety of GM1 bore the recognition site. Analysis of S20Y cell adherence to wells coated with derivatives of GM1 indicated that the cells did not adhere to asialo-GM1 and adherence to the methyl ester or de-N-acetyl derivatives was significantly reduced. Expression of the GM1 binding sites by S20Y cells appears to be density dependent; cells harvested at the confluent stage of growth were more adherent than those harvested at the preconfluent stage. Trypsin treatment of the S20Y and N115 cells resulted in a loss of binding to GM1-coated wells, suggesting that the cell surface GM1 binding site is a protein. In contrast, N115 cells showed no significant difference in their adherence to wells coated with GM1, GD1a, GT1b, Gal-Cer, asialo-GM1, or the methyl ester of GM1 when assayed under the same conditions as those imposed on the S20Y cells. The N115 cells did show a reduction in adherence to GM2-coated wells, suggesting that they recognized the terminal galactosyl moiety.

Galectin-1 Is a Major Receptor for Ganglioside GM1, a Product of the Growth-controlling Activity of a Cell Surface Ganglioside Sialidase, on Human Neuroblastoma Cells in Culture

“…Based on the application of the GM 1 -neoganglioprotein and the binding of cells to surfaceimmobilized GM 1 in the absence and presence of galectin-1-specific antibody, as well as the total number of immunologically assessed galectin-1 molecules and glycocytochemically quantitated neoganglioprotein-binding sites, it appears reasonable to conclude that the murine and the human lines differ in their GM 1 receptors. In this context it is remarkable that the murine adrenergic N115 neuroblastoma line failed to distinguish GM 1 from other gangliosides in a cell adherence assay, disclosing differences in the expression of ganglioside receptors even between two murine cell lines (62). Overall, our present results coalesce into the notion that galectin-1 is a major receptor for the carbohydrate portion of ganglioside GM 1 exposed on the surface of cultured human SK-N-MC neuroblastoma cells.…”

“…60). However, the enhancement of neuritogenesis and cell adherence by the sugar portion of GM 1 has been tentatively ascribed to the presence of a 71-kDa membrane protein in the case of murine cholinergic S20Y neuroblastoma cells (61)(62)(63). Based on the application of the GM 1 -neoganglioprotein and the binding of cells to surfaceimmobilized GM 1 in the absence and presence of galectin-1-specific antibody, as well as the total number of immunologically assessed galectin-1 molecules and glycocytochemically quantitated neoganglioprotein-binding sites, it appears reasonable to conclude that the murine and the human lines differ in their GM 1 receptors.…”

Review

“…Employing neoglycoconjugates with lysogangliosidesorsialyllactose, presence of sialic acid-binding receptor sites in the nervous system, blood cells, granulosa cells and tumours has been demonstrated (Tiemeyeref a/., 1989, Brought to you by | University of California Authenticated Download Date | 8/24/15 8:00 AM 1990; Gabius and Gabius, 1991;Kayserefa/., 1992;Hattori ei a/., 1995;Timoshenkoef a/., 1996). Independently, the monitoring of binding of gangliosides to neural or muscle cells with dependence on protein-carbohydrate recognition confirms the assumption that the sugar moiety of gangliosides displays ligand properties (Fueshko and Schengrund, 1990;Chan and Liu, 1991;Saqref a/., 1993). As already indicated with respect to l-type lectins, the sugar-binding event can be coupled with biosignalling, involving e.g.…”

Shedding and uptake of gangliosides and glycosylphosphatidylinositol-anchored proteins

“…Gangliosides added exogenously to epithelial cell cultures are taken up by the apical membranes, but do not pass the tight junction to the basolateral membranes of the cell [85]. Pretreatment of cells with trypsin reduces ganglioside uptake [86,87] and prevents adhesion of cells to GM1-coated wells [88]. The recovery of gangliosideuptake ability requires de novo synthesis of proteins [70].…”