Add time:08/21/2019         Source:sciencedirect.com

Gangliosides and extracellular matrix molecules influence neurite outgrowth, but the combinatorial effects of these endogenous agents on outgrowth are unclear. Exogenous gangliosides inhibit neurite outgrowth from SH-SY5Y cells stimulated with platelet-derived growth factor-BB, and different isoforms of the ceramide analog threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) stimulate (L-PDMP) or inhibit (D-PDMP) glycosphingolipid biosynthesis. In this study, we determined whether altering the endogenous ganglioside levels with PDMP in SH-SY5Y cells regulates neurite outgrowth on the outgrowth-supporting extracellular matrix molecule, laminin. In cells stimulated with 20 ng/ml platelet-derived growth factor-BB to promote outgrowth, we used image analysis to evaluate neurite outgrowth from SH-SY5Y cells grown on endogenous matrix or laminin and exposed to L- or D-PDMP. Both L- and D-PDMP decreased neurite initiation (the number of neurites/cell, the percent of neurite-bearing cells), elongation (the length of the longest neurite/cell, the total neurite length/cell), and branching (the number of branch points/neurite) from SH-SY5Y cells on endogenous matrix or laminin in a dose-dependent manner in serum-free or serum-containing medium. The inhibitory effects of each PDMP isoform were reversible. Inhibition of neurite outgrowth by L-PDMP could be mimicked by addition of exogenous gangliosides or C2-ceramide.Our analyses of neurite outgrowth in SH-SY5Y cells, a model of developing or regenerating noradrenergic neurons, demonstrate that increasing or decreasing endogenous ganglioside levels decreases neurite outgrowth. These results may indicate that SH-SY5Y cells undergo tight regulation by gangliosides, possibly through modulation of growth/trophic factor- and/or extracellular matrix-activated signaling cascades.

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