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. Authoritative and timely, Sphingosine-1-Phosphate: Methods and Protocols is a key resource for scientists working in this expanding and dynamic field.
1. Biochemical Methods for Quantifying Sphingolipids: Ceramide, Sphingosine, Sphingosine Kinase-1 Activity, and Sphingosine-1-Phosphate Brizuela Leyre and Cuvillier Olivier
2. Isoform-Selective Assays for Sphingosine Kinase Activity Melissa R. Pitman, Duyen H. Pham, and Stuart M. Pitson
3. Quantification of Sphingosine-1-Phosphate and Related Sphingolipids by Liquid Chromatography Coupled to Tandem Mass Spectrometry Constantin Bode and Markus H. Graler
4. A Clean-up Method for Mass Spectrometric Analysis of Sphingosine-1-Phosphate in Blood and Solid Tissues Using a Phosphate Capture Molecule Jun-ichi Morishige, Tamotsu Tanaka, and Kiyoshi Satouchi
5. Immunohistochemical Detection of Sphingosine-1-Phosphate and Sphingosine Kinase-1 in Human Tissue Samples Barbara Visentin, Gary Reynolds, and Roger Sabbadini
6. Assessment of Sphingosine-1-Phosphate Activity in Biological Samples by Receptor Internalization and Adherens Junction Formation Hideru Obinata and Timothy Hla
7. High Throughput Assays to Measure Intracellular Ca2+ Mobilization in Cells that Express Recombinant S1P Receptor Subtypes William J. Valentine and Gabor Tigyi
8. Quantifying Sphingosine-1-Phosphate-Dependent Activation of the RhoGTPases Matt D. Medlin, Joan M. Taylor, and Christopher P. Mack
9. Optimized Protocols to Analyze Sphingosine-1-Phosphate Signal Transduction Pathways During Acrosomal Exocytosis in Human Sperm Silvia A. Belmonte and Laila Suhaiman
10. Use of Intravital Microscopy and In Vitro Chemotaxis Assays to Study the Roles of Sphingosine-1-Phosphate in Bone Homeostasis Taeko Ishii, Shunsuke Kawamura, Issei Nishiyama, Junichi Kikuta, and Masaru Ishii
11. 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, and Timothy E. Kennedy
12. Sphingosine-1-Phosphate Signaling in Skeletal Muscle Cells Elisabetta Meacci, Francesca Bini, and Chiara Battistini
13. Maintenance of Human Embryonic Stem Cells by Sphingosine-1-Phosphate and Platelet-Derived Growth Factor Raymond C.B. Wong, Martin F. Pera, and Alice Pebay
14. Ceramide and Sphingosine-1-Phosphate Signaling in Embryonic Stem Cell Differentiation Erhard Bieberich
15. Sphingosine-1-Phosphate: An Alternative to Serum for Culture and Multipotency Maintenance of Adult Human Mesenchymal Stem Cells He Xiaoli, Sathish Jean, and Alirio J. Melendez
16. Sphingosine-1-Phosphate Signaling in Neural Progenitors Phillip Callihan and Shelley B. Hooks
17. Utilizing Sphingosine-1-Phosphate to Stimulate Sprouting Angiogenesis Shih-Chi Su and Kayla J. Bayless