Copyright Page	5
	DEDICATION	6
	PREFACE	7
	ABOUT THE EDITORS...	8
	ABOUT THE EDITORS...	9
	PARTICIPANTS	10
	Table of Contents	15
	ACKNOWLEDGEMENTS	21
	Chapter 1 An Overview of SphingolipidMetabolism:From Synthesis to Breakdown	22
	Sphingolipid Properties in Membranes	23
	De Novo Synthesis in the ER	24
	Serine Palmitoyltransferase and 3-Ketodihydrosphingosine Reductase	25
	Dihydroceramide Synthases/Ceramide Synthases and Dihydroceramide Desaturase	26
	Ceramide Transport from the ER to the Golgi	27
	Synthesis of Complex Sphingolipids	28
	Ceramide Galactosyltransferase and Galactosphingolipids	28
	Glucosylceramide Synthase and Derivatives of Glucosylceramide	29
	Sphingomyelin Synthesis	29
	Ceramide Kinase and Ceramide-1-Phosphate	30
	Catabolizing Complex Sphingolipids and Sphingomyelins into Ceramide	31
	The Catabolism of Ceramides and the Final Common Breakdown Pathway	33
	Acid, Neutral and Alkaline Ceramidases	33
	Sphingosine-1-Phosphate and Sphingosine Kinases 1 and 2	35
	Lipid Phosphate Phosphatases, S1P Phosphatases and the Salvage Pathway	36
	S1P Lyase in the Removal of Sphingoid Bases	37
	Conclusion	38
	References	38
	Chapter 2 Sphingolipid Transport	45
	Introduction	45
	Intramembrane Sphingolipid Movements	46
	Lateral Diffusion and Lateral Phase Separation of Sphingolipids	46
	Sphingolipid Raft Dynamics	47
	Transbilayer Transport	48
	Transbilayer Transfer of Sphingomyelin and Complex Glycosphingolipids	49
	Transbilayer Transfer of Monohexosylsphingolipids	52
	Transbilayer Transfer of Ceramide and Sphingoid Bases	52
	Transbilayer Transfer of Sphingosine-1-Phosphate	53
	Intermembrane Sphingolipid Transport	53
	Protein-Mediated Sphingolipid Transport	54
	CERT-Mediated Transport of Ceramides	54
	FAPP2-Mediated Transport of Glucosylceramide	57
	Glycolipid Transfer Proteins	57
	Membrane Contacts	58
	Sphingolipid Vesicular Transport	58
	Biosynthetic Vesicular Pathway	58
	Endocytic Pathway	59
	Conclusion	60
	References	61
	Chapter 3 Sphingolipid Analysisby High Performance LiquidChromatography-Tandem MassSpectrometry (HPLC-MS/MS)	67
	Introduction	67
	Sphingolipids: Structure and Composition	68
	LC-MS Methods for Detection and Analysis of Bioactive Sphingolipids	70
	Lipidomic Approach	70
	Sample Preparation	70
	Analysis of Intact Sphingolipids by Mass Spectrometry	71
	Mechanism of Electrospray Ionization Mass Spectrometry (ESI/MS)	72
	MS Scan Modes	72
	Specific Scan Modes for MS/MS Instrumentation	73
	Product Ion Scan	73
	Neutral Loss (NL)	73
	Precursor Ion Scan (PI)	73
	Multiple Reaction Monitoring (MRM)	73
	Sphingolipid Identification	73
	HPLC-MS/MS Methodology	74
	Quantitation	75
	Selection of Internal Standards (ISs)	76
	Quantitative Calibration	76
	Data Handling	76
	Alternative Methodology	77
	Conclusion	77
	References	77
	Chapter 4 Ceramide Synthases:Roles in Cell Physiology and Signaling	81
	Introduction	81
	Fatty Acid Specificity, Kinetics and Tissue Distribution	83
	Inhibitors	84
	Fumonisins	84
	Australifungin	85
	FTY720	85
	Posttranslational Modifications	85
	Membrane Topology	85
	Why Are There So Many Mammalian CerS?	87
	Roles of CerS in Signal Transduction and Disease	88
	Conclusion	89
	References	89
	Chapter 5 Tales and Mysteries of the EnigmaticSphingomyelin Synthase Family	93
	Sphingomyelin Biosynthesis: An Historical Perspective	93
	Initial Milestones	93
	Localization of SM Synthase Activity in Cells	94
	Discovery of a Ceramide Transfer Protein with a Key Role in SM Biosynthesis	96
	Alternative Pathways of SM Biosynthesis and Analogous Reactions	96
	Physicochemical Properties of SM	97
	The Multigenic Sphingomyelin Synthase (SMS) Family	97
	SMS Cloning Strategies	97
	Structural Organization and Reaction Chemistry of SMS Family Members	98
	SMS Family Members Display Striking Variations in Substrate Specificity	99
	Differential Expression of SMS1 and SMS2	100
	Cellular Functions of SMS Family Members	100
	SMS1 and SMS2 as Regulators of SM Homeostasis and Receptor-Mediated Signaling	100
	SMS1 and SMS2 as Regulators of Lipid-Based Signaling	101
	Conclusion	102
	References	103
	Chapter 6 Ceramide in Stress Response	107
	Introduction	107
	Chemical Structure and Biophysical Properties of Ceramide	108
	Changes in Ceramide Mass during Stress	108
	Mechanisms for Ceramide Generation during Stress	112
	Role of the De Novo Pathway for Ceramide Generation in Cellular Stress Response	112
	Heat Stress	113
	Septic Shock	113
	Lipotoxicity and Insulin Desensitization	114
	Programmed Cell Death	114
	Autophagy	115
	Mechanisms of Activation of De Novo Synthesis of Ceramide during Stress	115
	Role of the Sphingomyelinases in Cellular Stress Response	115
	Neutral Sphingomyelinase	116
	Hepatic Acute Phase Response	116
	Vascular Inflammation	116
	Apoptosis	116
	Growth Arrest	116
	Aging and Cancer	117
	Mechanisms of Activation of NSMase	117
	Acid Sphingomyelinase	118
	Endotoxic Shock	118
	Apoptosis (reviewed in ref. 162)	118
	Viral and Bacterial Infections	118
	Mechanisms of Activation of ASMase	118
	Evidence for a Coordinated Regulation of Multiple Pathways for Ceramide Generation	118
	Mechanisms of Ceramide Effects on Cellular Functions	119
	Ceramide-Interacting Molecules	119
	PKC (reviewed in ref. 114)	119
	PP2A (reviewed in ref. 181)	119
	Cathepsin D	119
	Indirect Targets of Ceramide	120
	Modulators of Apoptosis (reviewed in ref. 184)	120
	Regulators of Cell Cycle	120
	Regulators of Inflammation	120
	Ceramide Effects on Membrane Organization	121
	Conclusion	121
	References	122
	Chapter 7 Animal Models for Studyingthe Pathophysiology of Ceramide	130
	Introduction	130
	Sphingosine Kinase 1/2	130
	Ceramidases	131
	Acid Ceramidase	131
	Neutral Ceramidase	132
	Sphingomyelinases (SMase) and Sphingomyelin Synthases (SMS)	132
	Acid Sphingomyelinase (ASMase)	132
	Neutral Sphingomyelinase (nSMase) 1/2	133
	Sphingomyelin Synthases (SMS)	133
	S1P Lyase	133
	The Other GEM for Sphingolipid-Related Enzymes	134
	Conclusion	134
	References	135
	Chapter 8 Ceramide-1-Phosphate in Cell Survivaland Inflammatory Signaling	139
	Introduction	139
	Ceramide-1-Phosphate Synthesis and Degradation	140
	Ceramide-1-Phosphate: A Key Regulator of Cell Growth and Survival	142
	Ceramide-1-Phosphate and the Control of Inflammatory Responses	144
	Ceramide-1-Phosphate Mediates Macrophage Migration	145
	Conclusion	146
	References	147
	Chapter 9 Ceramide-1-Phosphate in Phagocytosisand Calcium Homeostasis	152
	Ceramide-1-Phosphate in Phagocytosis	152
	Ceramide-1-Phosphate as a Regulator of Calcium Homeostasis	155
	Conclusion	158
	References	159
	Chapter 10 Extracellular and Intracellular Actionsof Sphingosine-1-Phosphate	162
	Introduction	162
	Sphingolipid Metabolism	162
	Sphingosine Kinases	163
	SphK1	164
	SphK2	164
	SphK1 vs. SphK2	164
	S1P Receptors	166
	S1P1	166
	S1P2	167
	S1P3	167
	S1P4 and S1P5	167
	Evidence for Intracellular Targets of S1P	168
	S1P in Saccharomyces cerevisiae	168
	S1P in Arabidopsis thaliana	168
	S1P in Mammalian Cells	169
	Implications, Future Directions, and Conclusion	171
	References	171
	Chapter 11 Glucosylceramide in Humans	177
	Introduction	177
	Glucosylceramide Synthesis and Degradation	177
	Multiple Functions of Glucosylceramide	179
	Template for Higher Order Glycosphingolipids	179
	Membrane and Lipid Raft Constituent	179
	Cellular Protection in the Skin	180
	Cellular Protection in the Cardiovasculature	180
	Cellular Protection in the Brain	181
	Cellular Protection in the Immune System	181
	Cellular Protection in Carcinomas	182
	Conclusion	182
	References	183
	Chapter 12 Gangliosides as Regulators of CellMembrane Organization and Functions	186
	Introduction	186
	Segregation of Membrane Lipids and Detergent-Resistant Membrane Domains	190
	Lipid Membrane Domain Functions	192
	Gangliosides and Lipid Membrane Domains in the Nervous System	192
	The Glycosynapse	193
	GM3 and EGF Receptor	194
	GM3, Caveolae and the Regulation of Insulin Receptor and PDGF Receptor	195
	The Regulation of Glycosphingolipid Composition of the Plasma Membranes	195
	Conclusion	197
	References	197
	Chapter 13 Cancer Treatment Strategies TargetingSphingolipid Metabolism	206
	Introduction	206
	Sphingolipid Metabolism	207
	Ceramide Generated via Different Biochemical Routes Can Induce Apoptosis	208
	Ceramide as a Mediator of Cell Death by Chemopreventive Agents	209
	Ceramide Influences Both the Intrinsic and Extrinsic Apoptotic Pathways	209
	Sphingosine-1-Phosphate as a Counterbalance to Ceramide	210
	Inhibitory Effects of S1P on Apoptotic Pathways	211
	Sphingolipids Regulate Key Signaling Pathways That Control Cell Fate	211
	Sphingolipids and Autophagy	213
	Other Signaling Pathways Influenced by Sphingolipids	214
	Ceramide Regulates Cell Cycle Progression	214
	Ceramide and Telomerases	215
	Ceramide and S1P in Cancer Stem Cells	215
	Effects of S1P on Migration and Metastasis	215
	Cancer Cells Exhibit Molecular and Genetic Changes in Sphingolipid Metabolism	215
	Targeting Sphingolipids for Cancer Therapy	217
	S1P Signaling to Protect Normal Tissues from Therapy-Related Cytotoxicity	219
	Conclusion	219
	References	219
	Chapter 14 Therapeutic Strategies for Diabetesand Complications:A Role for Sphingolipids?	227
	Diabetes and Insulin Resistance	227
	Insulin Resistance and Altered Sphingolipid Metabolism	228
	Diabetic Pancreatic Dysfunction and Sphingolipids	230
	Diabetic Cardiovascular Dysfunction and Sphingolipids	230
	Diabetic Nephropathy and Sphingolipids	231
	Diabetic Retinopathy and Sphingolipids	231
	Therapeutics That Target Sphingolipid Metabolism or Sphingolipid Signaling in Diabetes	232
	Conclusion	233
	References	234
	Chapter 15 Roles for Sphingolipidsin Saccharomyces cerevisiae	238
	Introduction	238
	Sphingolipid Metabolism in S. cerevisiae	238
	Membrane-Associated Functions and Processes	240
	Signal Transduction Pathways That Require Sphingolipids	243
	Longevity and Cellular Aging	245
	Regulation of Sphingolipid Biosynthesis	246
	Conclusion and Future Developments	247
	References	248
	Chapter 16 Sphingolipid Signaling in FungalPathogens	253
	Sphingolipid Synthesis	253
	Cryptococcus Neofomans: Model of Sphingolipid Signaling in Fungi	254
	Sphingolipid Signaling in Other Pathogenic Fungi	256
	Conclusion	256
	References	257
	Chapter 17 Sphingolipids in Parasitic Protozoa	259
	Introduction	259
	Leishmania	259
	SL Pathway Genetics	261
	SL Salvage by Amastigotes	261
	Inhibition of SL Synthetic Pathways	262
	Trypanosoma brucei (ssp) and Trypanosoma cruzi	262
	Trypanosoma brucei	262
	Trypanosoma cruzi	263
	Trypanosomatid Sphingolipid Synthases	264
	Plasmodium falciparum	265
	Toxoplasma gondii	265
	Trichomonas vaginalis and Giardia lamblia	266
	Conclusion	266
	References	267
	Chapter 18 Biosynthesis of Sphingolipids in Plants(and Some of Their Functions)	270
	Introduction	270
	Pathway of Plant Sphingolipid Biosynthesis	271
	Functional Characterization of Genes and Enzymes Involved in Plant Sphingolipid Biosynthesis (2004-2008)	276
	Serine Palmitoyltransferases	276
	Long-Chain Base C4-Hydroxylases	277
	Ceramidase	277
	Fatty Acyl a-Hydroxylase	277
	Sphingolipid- 4(E)-Desaturase	278
	Sphingolipid- 8(E/Z)-Desaturases	278
	Inositolphosphorylceramide Synthase (IPCS)	278
	Long-Chain Base Kinase and Long-Chain Base Phosphate Phosphatase	279
	Long-Chain Base Phosphate Lyase	281
	Conclusion	281
	References	282
	Chapter 19 Computational Analysis of SphingolipidPathway Systems	285
	Introduction	285
	Sphingolipid Models and Their Potential Uses	289
	Conclusion	294
	References	295
	Appendix Introduction to Tools and Techniquesfor Ceramide-Centered Research	297
	Lipid Extraction	297
	Identification and Quantification of Steady State Levels of Ceramide	297
	Analysis of Ceramide Metabolism	299
	The Use of Ceramide Analogues	300
	Short-Chain Ceramides	300
	Fluorescent Ceramide Analogues	300
	Pharmacological Tools	300
	Genetic Tools	300
	RNA Interference	300
	Knockout Mice	300
	Conclusion	303
	References	303
	Index	307