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Volume 17, Number 18,
Issue of September 15, 1997
pp. 6952-6960
Copyright ©1997 Society for Neuroscience
Involvement of Sphingosine 1-Phosphate in Nerve Growth
Factor-Mediated Neuronal Survival and Differentiation
Received March 7, 1997; revised June 12, 1997; accepted June 27, 1997.
Lisa Cseh Edsall,
Grisha G. Pirianov, and
Sarah Spiegel
Department of Biochemistry and Molecular Biology, Georgetown
University Medical Center, Washington, DC 20007
Sphingolipid metabolites, such as ceramide and
sphingosine-1-phosphate (SPP), are emerging as a new class of second
messengers involved in cellular proliferation, differentiation, and
apoptosis. Nerve growth factor (NGF), a neurotrophic factor for
pheochromocytoma PC12 cells, induced a biphasic increase in the
activity of sphingosine kinase, the enzyme that catalyzes the formation
of SPP. This activation was blocked by K252a, an inhibitor of tyrosine
kinase A (trkA). A rapid 1.7-fold increase was followed by a marked
prolonged increase reaching a maximum of fourfold to fivefold
stimulation with a concomitant increase in SPP levels and a
corresponding decrease in endogenous sphingosine levels. Levels of
ceramide, the precursor of sphingosine, were only slightly decreased by
NGF in serum-containing medium. However, NGF decreased the elevation of
ceramide induced by serum withdrawal. Treatment of PC12 cells with SPP
did not induce neurite outgrowth or neurofilament expression, yet it
enhanced neurofilament expression elicited by suboptimal doses of NGF. Moreover, SPP also protected PC12 cells from apoptosis induced by serum
withdrawal. To further substantiate a role for SPP in the
cytoprotective actions of NGF, we found that
N,N-dimethylsphingosine, a competitive inhibitor of
sphingosine kinase, also induced apoptosis and interfered with the
survival effect of NGF. These effects were counteracted by exogenous
SPP. Moreover, other structurally related compounds, such as
dihydrosphingosine 1-phosphate and lysophosphatidic acid, had no
significant protective effects. Our results suggest that activation of
sphingosine kinase and subsequent formation of SPP may play an
important role in the differentiation and survival effects induced by
NGF.
Key words:
NGF;
sphingolipid metabolites;
sphingosine 1-phosphate;
apoptosis;
neuronal differentiation;
trkA;
signal transduction
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