Abstract
Sphingosine-1-phosphate (S1P) and its receptors are important in nervous system development. Reliable in vitro human model systems are needed to further define specific roles for S1P signaling in neural development. We have described S1P-regulated signaling, survival, and differentiation in a human embryonic stem cell-derived neuroepithelial progenitor cell line (hNP1) that expresses functional S1P receptors. These cells can be further differentiated to a neuronal cell type and therefore represent a good model system to study the role of S1P signaling in human neural development. The following sections describe in detail the culture and differentiation of hNP1 cells and two assays to measure S1P signaling in these cells.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Pitson SM, Pebay A (2009) Regulation of stem cell pluripotency and neural differentiation by lysophospholipids. Neurosignals 17:242–254
McGiffert C, Contos JJ, Friedman B, Chun J (2002) Embryonic brain expression analysis of lysophospholipid receptor genes suggests roles for s1p(1) in neurogenesis and s1p(1-3) in angiogenesis. FEBS Lett 531:103–108
Mizugishi K, Yamashita T, Olivera A, Miller GF, Spiegel S, Proia RL (2005) Essential role for sphingosine kinases in neural and vascular development. Mol Cell Biol 25:11113–11121
Blanc CA, Grist JJ, Rosen H, Sears-Kraxberger I, Steward O, Lane TE (2015) Sphingosine-1-phosphate receptor antagonism enhances proliferation and migration of engrafted neural progenitor cells in a model of viral-induced demyelination. Am J Pathol 185:2819–2832
Miron VE, Schubart A, Antel JP (2008) Central nervous system-directed effects of FTY720 (fingolimod). J Neurol Sci 274:13–17
Kimura A, Ohmori T, Ohkawa R, Madoiwa S, Mimuro J, Murakami T, Kobayashi E, Hoshino Y, Yatomi Y, Sakata Y (2007) Essential roles of sphingosine 1-phosphate/S1P1 receptor axis in the migration of neural stem cells toward a site of spinal cord injury. Stem Cells 25:115–124
Harada J, Foley M, Moskowitz MA, Waeber C (2004) Sphingosine-1-phosphate induces proliferation and morphological changes of neural progenitor cells. J Neurochem 88:1026–1039
Guo H, Zhao Z, Yang Q, Wang M, Bell RD, Wang S, Chow N, Davis TP, Griffin JH, Goldman SA (2013) An activated protein C analog stimulates neuronal production by human neural progenitor cells via a PAR1-PAR3-S1PR1-Akt pathway. J Neurosci 33:6181–6190
Dhara SK, Hasneen K, Machacek DW, Boyd NL, Rao RR, Stice SL (2008) Human neural progenitor cells derived from embryonic stem cells in feeder-free cultures. Differentiation 76:454–464
Hurst JH, Mumaw J, Machacek DW, Sturkie C, Callihan P, Stice SL, Hooks SB (2008) Human neural progenitors express functional lysophospholipid receptors that regulate cell growth and morphology. BMC Neurosci 9:118
Callihan P, Zitomer NC, Stoeling MV, Kennedy PC, Lynch KR, Riley RT, Hooks SB (2012) Distinct generation, pharmacology, and distribution of sphingosine 1-phosphate and dihydrosphingosine 1-phosphate in human neural progenitor cells. Neuropharmacology 62:988–996
Callihan P, Ali MW, Salazar H, Quach N, Wu X, Stice SL, Hooks SB (2014) Convergent regulation of neuronal differentiation and Erk and Akt kinases in human neural progenitor cells by lysophosphatidic acid, sphingosine 1-phosphate, and LIF: specific roles for the LPA1 receptor, ASN Neuro 6(6):1–18
Tuggle K, Ali M, Salazar H, Hooks S (2014) Regulator of G protein signaling transcript expression in human neural progenitor differentiation: R7 subfamily regulation by DNA methylation. Neurosignals 22:43–51
Hettinger-Smith BD, Leid M, Murray TF (1996) Chronic exposure to adenosine receptor agonists and antagonists reciprocally regulates the A1 adenosine receptor-adenylyl cyclase system in cerebellar granule cells. J Neurochem 67:1921–1930
Salomon Y (1979) Adenylate cyclase assay. Adv Cyclic Nucleotide Res 10:35–55
Hepler JR, Nakahata N, Lovenberg TW, DiGuiseppi J, Herman B, Earp HS, Harden TK (1987) Epidermal growth factor stimulates the rapid accumulation of inositol (1,4,5)-trisphosphate and a rise in cytosolic calcium mobilized from intracellular stores in A431 cells. J Biol Chem 262:2951–2956
Shin S, Mitalipova M, Noggle S, Tibbitts D, Venable A, Rao R, Stice SL (2006) Long-term proliferation of human embryonic stem cell-derived neuroepithelial cells using defined adherent culture conditions. Stem Cells 24:125–138
Dhara SK, Stice SL (2008) Neural differentiation of human embryonic stem cells. J Cell Biochem 105:633–640
Dan H, Sanes TAR, Harris WA (2006) Axon growth and guidance. Development of the nervous system, 2nd edn. Elsevier Academic Press, London, UK
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media New York
About this protocol
Cite this protocol
Callihan, P., Alqinyah, M., Hooks, S.B. (2017). Sphingosine-1-Phosphate (S1P) Signaling in Neural Progenitors. In: Pébay, A., Turksen, K. (eds) Sphingosine-1-Phosphate. Methods in Molecular Biology, vol 1697. Humana Press, New York, NY. https://doi.org/10.1007/7651_2017_3
Download citation
DOI: https://doi.org/10.1007/7651_2017_3
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7412-2
Online ISBN: 978-1-4939-7413-9
eBook Packages: Springer Protocols