Cover -- Title page -- Copyright page -- Contents -- Preface -- Contributors -- CHAPTER 1: Lysophosphatidic Acid (LPA) Receptor Signaling -- 1.1. Introduction -- 1.2. LPA Metabolism -- 1.3. Autotaxin -- 1.4. LPA Receptors -- 1.4.1. LPA1 -- 1.4.2. LPA2 -- 1.4.3. LPA3 -- 1.4.4. LPA4 -- 1.4.5. LPA5 -- 1.4.6. LPA6 -- 1.5. LPA Receptor Agonists and Antagonists -- References -- CHAPTER 2: Sphingosine 1-Phosphate (S1P) Receptors -- 2.1. Introduction -- 2.2. S1P Metabolism/Enzyme, and Transport -- 2.2.1. S1P Metabolism and Enzymes -- 2.2.2. Sphingosine Kinases -- 2.2.3. S1P Phosphatases and S1P Lyase -- 2.3. S1P Receptor Subtypes, and Physiological Functions -- 2.3.1. S1P1 -- 2.3.2. S1P2 -- 2.3.3. S1P3 -- 2.3.4. S1P4 -- 2.3.5. S1P5 -- 2.4. Concluding Remarks -- References -- CHAPTER 3: Global Gene Expression Program of Lysophosphatidic Acid (LPA)-Stimulated Fibroblasts -- 3.1. Introduction -- 3.2. The Global Transcriptional Response of MEFs to LPA -- 3.3. Upregulated Genes -- 3.4. Downregulated Genes -- 3.5. Induction of Genes that Encode Secreted Factors -- 3.6. Overlap between the Expression Profiles of LPA and EGF -- 3.7. Conclusions -- Acknowledgments -- References -- CHAPTER 4: Identification of Direct Intracellular Targets of Sphingosine 1-Phosphate (S1P) -- 4.1. Introduction -- 4.2. Intracellular Targets for S1P -- 4.3. Methods to Identify Intracellular S1P Targets -- 4.3.1. S1P Immobilized on Agarose Beads -- 4.3.2. Binding of 32P-Labeled S1P to Targets -- 4.3.3. Mass Measurement of Endogenous S1P in Immunoprecipitates of Target Proteins -- 4.4. Other Potentially Useful Methods to Identify Lipid Binding Proteins -- 4.4.1. Lipid Strips for Identification of Binding Proteins (Protein-Lipid Overlay) -- 4.4.2. Detection of Lipid Binding Proteins by Enzyme-Linked Immunadsorbent Assays -- 4.4.3. Liposome Pull Down -- 4.5. Concluding Remarks.

Acknowledgments -- References -- CHAPTER 5: Lysophospholipid Receptor Signaling Platforms: The Receptor Tyrosine Kinase-G Protein-Coupled Receptor Signaling Complex -- 5.1. Introduction -- 5.2. Lysophospholipid Receptor-Receptor Tyrosine Kinase Complexes -- 5.3. Other Lysophospholipid Receptor Signaling Platforms -- 5.4. Other Examples of RTK-GPCR Signaling Platforms -- 5.5. Interaction of RGS Proteins with Receptor Tyrosine Kinase-Lysophospholipid Receptor Signaling Complexes -- 5.6. S1P and RTK Transactivation -- 5.7. Approaches for the Study of Receptor Tyrosine Kinase-Lysophospholipid Receptor Signaling Complexes -- 5.8. Some Useful Protocols for Studying RTK-Lysophospholipid Receptor Signaling Platforms -- 5.8.1. Compounds -- 5.8.2. Cells -- 5.8.3. Immunoprecipitation -- 5.8.4. Immunofluorescence -- 5.8.5. GTP-γ-S Binding Assay -- 5.8.6. Cell Migration -- 5.9. Conclusion -- Acknowledgment -- References -- CHAPTER 6: Autotaxin: A Unique Ecto-Type Pyrophosphodiesterase with Diverse Functions -- 6.1. History of Autotaxin (ATX) -- 6.2. Structure of ATX -- 6.3. Expression of ATX -- 6.4. ATX Knockout Mice and Transgenic Mice -- 6.5. Role of ATX in Blood Vessel Formation -- 6.6. Role of ATX in Cancer -- 6.7. Clinical Aspects of ATX -- 6.8. ATX Inhibitors -- 6.9. Autotaxin Research Perspectives -- References -- CHAPTER 7: Studies on Autotaxin Signaling in Endocytic Vesicle Biogenesis and Embryonic Development Using Whole Embryo Culture and Electroporation -- 7.1. Introduction -- 7.2. Results and Discussion -- 7.2.1. Lysosomes Are Fragmented in the Yolk Sac VE Cells of Enpp2-/- Embryos -- 7.2.2. WEC Is Useful to Dissect Signaling Pathways Involved in the Defects of Enpp2-/- Embryos -- 7.2.3. LPA Receptors and Rho-ROCK-LIM Kinase (LIMK) Signaling Are Required for Lysosome Biogenesis -- 7.2.4. Autotaxin Regulates Actin Turnover Dynamics.

7.2.5. Electroporation-Mediated Inhibition of the Rho-ROCK-LIMK-Cofilin Pathway Induces Lysosome Defects in VE Cells -- 7.2.6. Electroporation-Mediated Activation of the Rho-ROCK-LIMK-Cofilin Pathway Induces Lysosome Defects in VE Cells -- 7.2.7. Head Cavity Is Formed by the Blockade of Rho-ROCK Signaling -- 7.2.8. Yolk Sac VE Cells Provide a Unique Opportunity to Study Vesicle Trafficking -- 7.3. Methods -- 7.3.1. Generation of Enpp2-Deficient Mice -- 7.3.2. Histological Analysis -- 7.3.3. Imaging of Lysosomes -- 7.3.4. Ex Vivo WEC -- 7.3.5. Electroporation of the Yolk Sac -- Acknowledgments -- References -- CHAPTER 8: Standardization and Quantification of Lysophosphatidic Acid Compounds by Normal-Phase and Reversed-Phase Chromatography-Tandem Mass Spectrometry -- 8.1. Preparation and Handling of Lysophosphatidic Acid (LPA) Compounds -- 8.2. 1H and 31P NMR Characterization -- 8.3. LC/MS/MS of LPA Compounds -- 8.3.1. Reversed-Phase UPLC/MS/MS -- 8.3.2. Normal-Phase LC/MS/LIT -- 8.4. Discussion -- Acknowledgments -- References -- CHAPTER 9: Sphingosine Kinases: Biochemistry, Regulation, and Roles -- 9.1. Introduction -- 9.2. SphK Structure, Isoforms, and Characteristics -- 9.2.1. Structural Insights from Bacterial Homologs -- 9.2.2. Human SphK Isoforms -- 9.3. Physiologic and Pathophysiologic Roles of SphKs -- 9.4. Posttranslational Regulation of SphK1 -- 9.4.1. Activation of SphK1 by Phosphorylation -- 9.4.2. SphK1 Translocation to the Plasma Membrane -- 9.4.3. Activation of SphK1 by Protein-Protein Interactions -- 9.5. Posttranslational Regulation of SphK2 -- 9.6. Transcriptional Regulation of SphKs -- 9.7. Extracellular SphKs -- 9.8. SphK Inhibitors -- 9.9. Conclusions -- 9.10. Common SphK Methods -- 9.10.1. In Vitro Assay of SphK Activity -- 9.10.2. Assay of SphK Activity in Intact Cells -- References.

CHAPTER 10: Functional and Physiological Roles of Sphingosine 1-Phosphate Transporters -- 10.1. Introduction -- 10.2. Physiological Release of S1P -- 10.3. Involvement of ABC Transporters in S1P Release -- 10.3.1. ABCA1 -- 10.3.2. ABCC1 -- 10.3.3. ABCG2 -- 10.4. The Role of the Spns2 Transporter in S1P Release -- 10.5. Human SPNS2 Functions as an FTY720-P Transporter -- 10.6. Conclusion -- 10.7. Methods -- Acknowledgments -- References -- CHAPTER 11: Lipid Phosphate Phosphatases and Signaling by Lysophospholipid Receptors -- 11.1. Introduction and Historical Dimension -- 11.1.1. Structure of LPPs -- 11.1.2. Proteins Related to Mammalian LPPs -- 11.1.3. Measurement of PAP and LPP Activities -- 11.1.4. Role of the LPPs in the Degradation of Extracellular LPA -- 11.1.5. Role of the LPPs in the Degradation of Extracellular S1P -- 11.1.6. Role of the LPPs in the Degradation of Other Extracellular Lipid Phosphates -- 11.1.7. Noncatalytic Actions of LPPs on the Cell Surface -- 11.1.8. Intracellular Functions of LPPs -- 11.1.9. Animal Models of LPP Activity -- 11.2. Conclusions -- Acknowledgments -- References -- CHAPTER 12: Lipid Phosphate Phosphatases: Recent Progress and Assay Methods -- 12.1. Introduction -- 12.2. LPP Nomenclature, Gene Structures, and Expression Patterns -- 12.3. Physiological Functions of Mammalian LPP Enzymes Revealed by Gene Targeting Studies in Mice -- 12.3.1. LPP2/PPAP2C -- 12.3.2. LPP1/PPAP2A -- 12.3.3. LPP3/PPAP2B -- 12.4. Genetic Studies of LPPs in Nonmammalian Systems -- 12.4.1. LPPs in Fly Development -- 12.4.2. Germ Cell Migration in Flies -- 12.4.3. Role of LPPs in the Eye Phototransduction Cycle in Flies -- 12.4.4. Role of LPPs in Arabidopsis Injury Responses -- 12.5. Lipid Phosphatase-Related Proteins and PA Phosphatase Domain Containing Proteins.

12.5.1. PPAPDC Proteins Are Lipid Diphosphate Preferring Enzymes with Roles in Regulation of Cell Growth, Tumorigenesis, and Differentiation -- 12.5.2. Lipid Phosphatase-Related Proteins Are Regulators of Cell Morphology -- 12.6. Concluding Comments -- 12.7. Determination of LPP Activity -- 12.7.1. Quantitation of Lipid Phosphate Substrates -- 12.7.2. Determination of LPP Activity -- 12.7.3. Quantitation of LPP Substrates and Dephosphorylation Products by HPLC ESI MS/MS -- 12.7.4. Measurement of Fluorescent Lipid Uptake by Cultured Cells -- Acknowledgments -- References -- CHAPTER 13: Lysophosphatidic Acid (LPA) Signaling and Cardiovascular Pathology -- 13.1. Introduction -- 13.2. Circulating LPA Levels -- 13.2.1. Protocols for LPA Measurement in Plasma -- 13.3. LPA Signaling in Blood and Vascular Cells -- 13.4. Regulation of Blood Pressure -- 13.4.1. Protocol for Arterial Pressure Measurements -- 13.5. Blood Vessel and Lymphatic Formation -- 13.5.1. Protocol for Matrigel Angiogenesis Assay -- 13.6. Vascular Permeability -- 13.6.1. Vascular Permeability Protocol -- 13.7. Vascular Inflammation -- 13.7.1. Inflammation Models -- 13.8. Atherothrombosis -- 13.8.1. Experimental Models for Studying the Development of Intimal Hyperplasia and Atherosclerosis -- 13.9. Future Directions -- Acknowledgments -- References -- CHAPTER 14: Sphingosine 1-Phosphate (S1P) Signaling in Cardiovascular Physiology and Disease -- 14.1. Sphingosine 1-Phosphate (S1P) in Plasma: Sources and Carriers -- 14.2. S1P Producing and Degrading Enzymes in the Heart and the Vasculature -- 14.3. S1P Receptors in the Heart and Vessel Wall -- 14.4. S1P Signaling in Cardiac Development -- 14.5. S1P in Vascular Morphogenesis -- 14.6. S1P in Myocardial Reperfusion Injury and Preconditioning: S1P Receptors, Sphingosine Kinases, and S1P Lyase.

14.7. Control of Arterial Tone and Tissue Perfusion by S1P.