Sphingosine-1-Phosphate Methods and Protocols

Sphingosine-1-phosphate is a bioactive lysophospholipid which has become, in recent years, the focus of much research interest as it has widespread developmental and physio-pathological actions, controlling events within the nervous, reproductive, gastrointestinal, vascular, respiratory, and immune systems, in addition to having a prominent role in cancer, early mammalian embryogenesis, and stem cells.  In Sphingosine-1-Phosphate: Methods and Protocols, worldwide experts in the S1P field describe in-depth techniques in an array of cell types and with various physiological applications, showcasing the important effects of S1P in development and in physiopathology.  As a volume in the Methods in Molecular Biology™ series, chapters contain introductions to their respective topics, lists of the relevant materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls.

Biochemical Methods for Quantifying Sphingolipids: Ceramide, Sphingosine, Sphingosine Kinase-1 Activity, and Sphingosine-1-Phosphate.- Isoform-Selective Assays for Sphingosine Kinase Activity.- Quantification of Sphingosine-1-Phosphate and Related Sphingolipids by Liquid Chromatography Coupled to Tandem Mass Spectrometry.- A Clean-up Method for Mass Spectrometric Analysis of Sphingosine-1-Phosphate in Blood and Solid Tissues Using a Phosphate Capture Molecule.- Immunohistochemical Detection of Sphingosine-1-Phosphate and Sphingosine Kinase-1 in Human Tissue Samples.- Assessment of Sphingosine-1-Phosphate Activity in Biological Samples by Receptor Internalization and Adherens Junction Formation.- High Throughput Assays to Measure Intracellular Ca²⁺ Mobilization in Cells that Express Recombinant S1P Receptor Subtypes.- Quantifying Sphingosine-1-Phosphate-Dependent Activation of the RhoGTPases.- Optimized Protocols to Analyze Sphingosine-1-Phosphate Signal Transduction Pathways During Acrosomal Exocytosis in Human Sperm.- Use of Intravital Microscopy and In Vitro Chemotaxis Assays to Study the Roles of Sphingosine-1-Phosphate in Bone Homeostasis.- Assessment of Sphingosine-1-Phosphate Receptor Expression and Associated Intracellular Signaling Cascades in Primary Cells of the Human Central Nervous System.- Sphingosine-1-Phosphate Signaling in Skeletal Muscle Cells.- Maintenance of Human Embryonic Stem Cells by Sphingosine-1-Phosphate and Platelet-Derived Growth Factor.- Ceramide and Sphingosine-1-Phosphate Signaling in Embryonic Stem Cell Differentiation.- Sphingosine-1-Phosphate: An Alternative to Serum for Culture and Multipotency Maintenance of Adult Human Mesenchymal Stem Cells.- Sphingosine-1-Phosphate Signaling in Neural Progenitors.- Utilizing Sphingosine-1-Phosphate to Stimulate Sprouting Angiogenesis.