Assessment of sphingosine-1-phosphate receptor expression and associated intracellular signaling cascades in primary cells of the human central nervous system

Veronique E Miron, Bryce A Durafourt, Jack P Antel, Timothy E Kennedy

Research output: Contribution to journalArticlepeer-review

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.
Original languageEnglish
Pages (from-to)141-54
Number of pages14
JournalMethods in Molecular Biology
Volume874
DOIs
Publication statusPublished - 2012

Keywords

  • RNA, Messenger
  • Blotting, Western
  • Oligodendroglia
  • Cells, Cultured
  • Neurons
  • Humans
  • Receptors, Lysosphingolipid
  • Neural Stem Cells
  • Reverse Transcriptase Polymerase Chain Reaction
  • Central Nervous System
  • Signal Transduction

Fingerprint

Dive into the research topics of 'Assessment of sphingosine-1-phosphate receptor expression and associated intracellular signaling cascades in primary cells of the human central nervous system'. Together they form a unique fingerprint.

Cite this