Tenascin-C inhibits  1 integrin-dependent cell adhesion and neurite outgrowth on fibronectin by a disialoganglioside-mediated signaling mechanism

Abstract

We have previously shown that the extracellular matrix molecule tenascin-C inhibits fibronectin-mediated cell adhesion and neurite outgrowth by an interaction with a cellular RGDindependent receptor which interferes with the adhesion and neurite outgrowth promoting activities of the fibronectin receptor(s). Here we demonstrate that the inhibitory effect of tenascin-C on β 1 integrin-dependent cell adhesion and neurite outgrowth is mediated by the interaction of the protein with membrane-associated disialogangliosides, which interferes with protein kinase C-related signaling pathways. First, in substratum mixtures with fibronectin, an RGD sequencecontaining fragment of the molecule or synthetic peptide, tenascin-C inhibited cell adhesion and spreading by a disialoganglioside-dependent, sialidase-sensitive mechanism leading to an inhibition of protein kinase C. Second, the interaction of intact or trypsinized, i.e., cell surface glycoproteinfree, cells with immobilized tenascin-C was strongly inhibited by gangliosides or antibodies to gangliosides and tenascin-C. Third, preincubation of immobilized tenascin-C with soluble disialogangliosides resulted in a delayed cell detachment as a function of time. Similar to tenascin-C, immobilized antibody to GD2 (3F8) or sphingosine, a protein kinase C inhibitor, strongly inhibited RGD-dependent cell spreading. Finally, the degree of tenascin-C-induced inhibition of cell adhesion was proportional to the degree of disialoganglioside levels of expression by different cells suggesting the relevance of such mechanism in modulating integrin-mediated cell-matrix interactions during pattern formation or tumor progression.

Functional role of a glycolipid in directional movements of neurons

“…However, based on the data summarized in the previous sections and also in the literature (Cheresh 1987, Pande 2000, Armulik et al 1999, Iwabuchi et al 1998, Probstmeier and Pesheva 1999, Hakomori and Igarashi 1995, Lloyd and Furukawa 1998, Dulabon et al 2000, Boldt et al 1977, Tiemeyer et al 1989 it is possible to suggest at least three mechanisms to explain the role of gangliosides (Fig. 1).…”

Lactosylceramide Is Essential for the Osteoclastogenesis Mediated by Macrophage-Colony-stimulating Factor and Receptor Activator of Nuclear Factor-κB Ligand

“…54) and to modulate integrin-mediated adhesion (55,56). In neuronal cells, binding to disialogangliosides by tenascin or anti-GD2 antibody causes inhibition of protein kinase C and prevention of integrin-dependent adhesion to fibronectin, resulting in inhibition of neurite outgrowth (57,58). Therefore, GM1, which is not expressed in GM2/GD2-TϪ/Ϫ mice, increased after RANKL stimulation; this might be important to TRAP-positive mononuclear and multinuclear cell formation, especially for cell-to-cell attachment and fusion.…”

Myelin-associated Glycoprotein Interacts with Ganglioside GT1b

“…To examine whether GD1a and GT1b could compete with endogenous neuronal MAG receptors for MAG binding, we preincubated Fc protein with gangliosides prior to addition into neurite outgrowth assays (37). Preincubation of MAG-Fc with a mixture of brain gangliosides, purified GT1b or GD1a but not asialo GM1 or GM1, blocked the inhibitory action of MAG (Fig.…”

“…In neuronal cells, binding to disialogangliosides by tenascin-C, tenascin-R, or an anti-GD2 antibody causes inhibition of protein kinase C and prevention of integrin-dependent adhesion to fibronectin, resulting in inhibition of neurite outgrowth (37,67).…”