The Zebrafish: Disease Models and Chemical Screens

Part I: Adipose Tissue

Chapter 1. In vivo imaging and quantification of regional adiposity in zebrafish

• Introduction
• 1. Rationale
• 2. Materials
• 3. Methods
• 4. Summary

Part II: Innate and Adaptive Immune Systems

Chapter 2. Innate immune cells and bacterial infection in zebrafish

• Introduction
• 1. Quantifying the Innate Immune Cell Response to Bacterial Infection
• 2. Bioassays for Assessing Neutrophil Bactericidal Function
• 3. Bioassays for Assessing Macrophage Bactericidal Function
• 4. Drug Discovery Platform to Identify New Immunomodulatory Drugs

Chapter 3. Best practices for germ-free derivation and gnotobiotic zebrafish husbandry

• Introduction
• 1. Experimental Procedures
• 2. Prospectus
• 3. Solutions

Chapter 4. Infectious disease models in zebrafish

• Introduction
• 1.. Methods for Systemic Bacterial and Viral Infections
• 2.. Methods for Localized Bacterial and Viral Infections
• Conclusion

Chapter 5. Live imaging the earliest host innate immune response to preneoplastic cells using a zebrafish inducible KalTA4-ERT2/UAS system

• Introduction
• 1. Generation of Preneoplastic Cell Clones Using the KalTA4-ERT2/UAS System in Zebrafish Larvae
• 2. Live Imaging Preneoplastic Cell: Neutrophil Interaction Using Confocal Microscopy
• 3. Image Analysis and 4D Reconstruction
• Conclusion

Chapter 6. Studying the adaptive immune system in zebrafish by transplantation of hematopoietic precursor cells

• Introduction
• 1. Methodology for the Transplantation of Hematopoietic Cells
• 2. Discussion
• 3. Future Directions

Part III: Blood and Lymph

Chapter 7. Hematopoietic stem cell development: Using the zebrafish to identify extrinsic and intrinsic mechanisms regulating hematopoiesis

• Introduction to Hematopoietic Development
• 1. Use of Zebrafish to Investigate Hematopoietic Stem Cell Development
• 2. Zebrafish Tools and Protocols

Chapter 8. Studying disorders of vertebrate iron and heme metabolism using zebrafish

• 1. Overview of Vertebrate Cellular Iron and Heme Metabolism
• 2. Advantageous Properties of Zebrafish to Study Genetics
• 3. Tools to Study Iron and Heme Metabolism Using Zebrafish
• Conclusions and Future Directions

Chapter 9. The lymphatic vasculature revisited—new developments in the zebrafish

• 1. Development of the Lymphatic System in the Zebrafish Trunk
• 2. Molecular Mechanisms Regulating Lymphatic Cell Fate Specification
• 3. Vegfc Signaling and Sprouting From the Posterior Cardinal Vein
• 4. Parachordal Lymphangioblast Migration at the Level of the Horizontal Myoseptum
• 5. Development of Lymphatic Structures in the Head and the Gut
• Concluding Remarks

Part IV: Visceral Organs

Chapter 10. Modeling intestinal disorders using zebrafish

• 1. Intestinal Development, Morphology, and Physiology
• 2. Intestinal Microbiota and Host–Microbe Interactions
• 3. Intestinal Inflammatory Conditions
• 4. Enteric Nervous System and Motility Disorders
• 5. Intestinal Tumorigenesis and Cancer
• Concluding Remarks

Chapter 11. Analysis of pancreatic disease in zebrafish

• Introduction
• 1. Method 1. Fasting Adult Zebrafish
• 2. Method 2. Weighing Live, Swimming Zebrafish Without Anesthetic
• 3. Method 3. Glucose Delivery to Adult Zebrafish Using Intraperitoneal Injection
• 4. Method 4. Dissection of the Pancreas, En Bloc, for Histology
• 5. Method 5. Sterile Dissection and Culture of the Principal Islet

Part V: Musculoskeletal System

Chapter 12. Using the zebrafish to understand tendon development and repair

• Introduction
• 1. Tendon Structure
• 2. Tendon Formation and Differentiation
• 3. Tissue Interactions Within the Developing Musculoskeletal System
• 4. Methods to Study the Embryonic Tendon Program in Zebrafish
• Conclusion

Chapter 13. Small teleost fish provide new insights into human skeletal diseases

• Introduction
• 1. A Fresh View on the Teleost Skeleton and Its Special Characters
• 2. Analyzing Skeletal Phenotypes of Small Fish
• 3. Mutant and Transgenic Fish Open New Directions in Skeletal Research
• 4. Genetic Phenocopies of Human Skeletal Diseases
• 5. Lifelong Tooth Replacement
• Concluding Remarks

Chapter 14. Muscular dystrophy modeling in zebrafish

• Introduction
• 1. Muscular Dystrophies and the Dystrophin-Associated Glycoprotein Complex
• 2. Skeletal Muscle Properties in Zebrafish
• 3. Models of Muscular Dystrophy in Zebrafish
• 4. Small Molecule Screens
• Conclusions

Part VI: Central and Sensory Nervous Systems

Chapter 15. Analysis of myelinated axon formation in zebrafish

• Introduction
• 1. Visualization of Myelinating Glia in Zebrafish
• 2. Genetic Analysis of Myelin Development in Zebrafish
• 3. Pharmacological Manipulation of Myelinated Axons in Zebrafish
• 4. Plasticity, Maintenance, and Regeneration of Myelinated Axons in Zebrafish
• Conclusions

Chapter 16. Zebrafish models of human eye and inner ear diseases

• Introduction
• 1. Zebrafish Models of Eye Disease
• 2. Zebrafish Models of Ear Disease
• 3. Zebrafish Models of Syndromes Affecting Eye and/or Ear
• Conclusion

Part VII: Cancer

Chapter 17. A zebrafish xenograft model for studying human cancer stem cells in distant metastasis and therapy response

• Introduction
• 1. Establishment of Human Cancer Stem Cells Xenograft Model
• 2. Analysis of Interaction Between Cancer Stem/Progenitor-Like Cells and Microenvironment
• 3. Genetic and Chemical Targeting of Tumor-Stroma Interactions in Cancer Stem/Progenitor-Like Cells–Xenograft
• Conclusion

Chapter 18. Zebrafish as a model for von Hippel Lindau and hypoxia-inducible factor signaling

• Introduction
• 1. Hypoxic Signaling
• 2. Hypoxic Signaling: Overview of the Zebrafish Orthologues
• 3. The Zebrafish HIF Genes
• 4. HIF Protein Expression
• 5. The HIF Hydroxylases
• 6. The VHL Genes
• 7. HIF Binding Sites in the Genome
• 8. Hypoxic/HIF Target Gene Comparison
• 9. Hypoxic Signaling Reporters in Zebrafish
• 10. Links Between HIF and the Circadian Clock
• 11. Zebrafish VHL Mutants as Models of Hif Hyperactivation
• 12. Hematopoietic and Angiogenic Phenotypes in VHL Mutants
• 13. Vhl/HIF Effects on Metabolism
• 14. Kidney Defects in VHL Mutants and Cancer
• 15. Links Between VHL HIF and P53
• Appendix A. Supplementary data

Chapter 19. Discovering novel oncogenic pathways and new therapies using zebrafish models of sarcoma

• Introduction
• 1. Rhabdomyosarcoma
• 2. Malignant Peripheral Nerve Sheath Tumors
• 3. Ewing's Sarcoma
• 4. Chordoma
• 5. Hemangiosarcoma
• 6. Liposarcoma
• 7. Future Perspectives and Opportunities

Chapter 20. Zebrafish models of leukemia

• 1. T-Cell Acute Lymphoblastic Leukemia
• 2. B-Cell Acute Lymphoblastic Leukemia
• 3. Myeloid Malignancies
• Conclusions

Chapter 21. Investigating microglia-brain tumor cell interactions in vivo in the larval zebrafish brain

• Introduction
• 1. Methods
• 2. Summary

Part VIII: Transplantation

Chapter 22. Transplantation in zebrafish

• Introduction
• 1. Rationale
• 2. Methods
• 3. Discussion

Part IX: Chemical Screening

Chapter 23. Chemical screening in zebrafish for novel biological and therapeutic discovery

• Introduction
• 1. Rationale
• 2. Materials and Methods
• 3. Discussion/caveats
• Summary