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ARF6 Regulates Neuron Differentiation through Glucosylceramide Synthase

Journal article
Authors Lu Li
Marcus Ståhlman
Mikael Rutberg
Liliana Håversen
Per Fogelstrand
Linda Andersson
Malin Levin
Jan Borén
Published in Plos One
Volume 8
Issue 3
Pages Article Number: e60118
ISSN 1932-6203
Publication year 2013
Published at Institute of Medicine, Department of Molecular and Clinical Medicine
Pages Article Number: e60118
Language en
Keywords glycosphingolipid synthesis, neurite outgrowth, cells, inhibitor, arno, endocytosis, activation, transport, endosome, dynamics,
Subject categories Medical Biotechnology


The small GTPase ADP ribosylation factor 6 (ARF6) mediates endocytosis and has in addition been shown to regulate neuron differentiation. Here we investigated whether ARF6 promotes differentiation of Neuro-2a neuronal cells by modifying the cellular lipid composition. We showed that knockdown of ARF6 by siRNA in Neuro-2a cells increased neuronal outgrowth as expected. ARF6 knockdown also resulted in increased glucosylceramide levels and decreased sphingomyelin levels, but did not affect the levels of ceramide or phospholipids. We speculated that the ARF6 knockdown-induced increase in glucosylceramide was caused by an effect on glucosylceramide synthase and, in agreement, showed that ARF6 knockdown increased the mRNA levels and activity of glucosylceramide synthase. Finally, we showed that incubation of Neuro-2a cells with the glucosylceramide synthase inhibitor D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-PDMP) normalized the increased neuronal outgrowth induced by ARF6 knockdown. Our results thus show that ARF6 regulates neuronal differentiation through an effect on glucosylceramide synthase and glucosylceramide levels.

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