title: Assessment of sphingosine-1-phosphate receptor expression and associated intracellular signaling cascades in primary cells of the human central nervous system.
abstract: Measuring the effects of sphingosine-1-phosphate (S1P) receptor modulators on human primary neural cells is of particular interest given the recent application of these central nervous system-accessible agents to the treatment of neurodegenerative diseases, such as multiple sclerosis. Issues to consider in experimental studies include the ability of some of these modulators to bind multiple S1P receptor subtypes simultaneously, the nonspecificity of commercially available S1P receptor antibodies, and activation of multiple intracellular signaling cascades by a given S1P receptor. Here, we discuss how to assay S1P receptor expression and activation using multiple agonists/antagonists, by linking the results of real-time reverse transcriptase polymerase chain reaction with the assessment of intracellular signaling derived from Western blot analyses.
authors.affiliation: Centre for Multiple Sclerosis Research, Scottish Centre for Regenerative Medicine, The University of Edinburgh, Edinburgh, UK.
authors.forename: Veronique E
authors.initials: VE
authors.lastname: Miron
authors.forename: Bryce A
authors.initials: BA
authors.lastname: Durafourt
authors.forename: Jack P
authors.initials: JP
authors.lastname: Antel
authors.forename: Timothy E
authors.initials: TE
authors.lastname: Kennedy
mesh_terms: D015153:Blotting, Western; D002478:Cells, Cultured; D002490:Central Nervous System / Q000166:cytology*; D006801:Humans; D058953:Neural Stem Cells / Q000166:cytology / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D009836:Oligodendroglia / Q000166:cytology; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D049349:Receptors, Lysosphingolipid / Q000819:agonists / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction
chemical_list: D012333:RNA, Messenger; D049349:Receptors, Lysosphingolipid