This is the first in a two-volume, comprehensive treatment of the methodologies used in researching the Zebrafish, an emerging vertebrate model system. The text includes discussions on development, genetic methodologies, and model applications.
Key Features
* Details state-of-the-art zebrafish protocols in a single-source reference
* Presents methods and reagents in user-friendly format
* Delineates critical steps and pitfalls of he procedures
* Illustrates techniques with full-color plates
* Summarizes many new and interesting developmental mutants
* Includes appendices with strain information and a compendium of zebrafish World Wide Web sites
* Relevant to clinicians interested in vertebrate models of human congenital diseases
Contributors xiii(4) Preface xvii PART I Introduction 3(26)
1. Overview of the Zebrafish System 3(8) H. William Detrich, III Monte Westerfield Leonard I. Zon I. Introduction 3(1) II. History of the Zebrafish System and Its Advantages and Disadvantages 4(2) III. Cell and Developmental Biology, Organogenesis, and Human Disease 6(1) IV. Genetics and Genomics 7(1) V. Future Prospects 8(1) VI. Conclusion 8(1) References 8(3)
2. Cell Cycles and Development in the Embryonic Zebrafish 11(18) Donald A. Kane I. Introduction 11(1) II. Terminology and the Staging Series 12(1) III. The Zygote Period 12(2) IV. The Cleavage Period 14(2) V. The Blastula Period 16(8) VI. The Gastrula Period 24(1) VII. The Segmentation Period 24(1) References 25(4) PART II Cell Culture and General Methods 29(34)
3. Zebrafish Embryonal Cell Culture 29(10) Angela Helmrich David Barnes I. Introduction 29(1) II. General Cell Culture Considerations 30(1) III. Primary and Multipassage Zebrafish Embryo Cell Culture 31(2) IV. Transfection 33(1) V. Applications to Other Fish Models 34(1) VI. Conclusion 35(1) References 36(3)
4. Primary Fibroblast Cell Culture 39(7) Barry H. Paw Leonard I. Zon I. Introduction 39(1) II. Material and Methods 40(1) III. Results and Discussion 41(2) References 43(3)
5. Production of Haploid and Diploid Androgenetic Zebrafish (Including Methodology for Delayed in Vitro Fertilization) 46(17) Graham E. Corley-Smith Bruce P. Brandhorst Charline Walker John H. Postlethwait I. Introduction 46(2) II. Equipment and Materials 48(2) III. Methods 50(4) IV. Results and Discussion 54(4) V. Conclusions and Perspectives 58(1) References 59(4) PART III Gene Expression and Function in Development 63(96)
6. Analysis of Protein and Gene Expression 63(24) Trevor Jowett I. In Situ Hybridization to RNA and Immunolocalization of Proteins 63(1) II. Probe Synthesis 64(3) III. Fixation 67(1) IV. Hybridization to Whole-Mount Embryos 68(3) V. Immunolocalization of Probes 71(2) VI. Two-Color in Situ Hybridization 73(4) VII. Double-Fluorescent in Situ Hybridization 77(1) VIII. Simultaneous Localization of Transcription and Translation Gene Products 78(4) IX. Embedding and Sectioning Whole-Mount Embryos 82(1) X. Solutions and Reagents 83(1) References 84(3)
7. Strategies to Perturb Zebrafish Development 87(30) Matthias Hammerschmidt Patrick Blader Uwe Strahle I. Introduction 87(1) II. Administration of Reagents 88(2) III. Reagents 90(5) IV. Experiments to Modify the Activity of Specific Gene Products 95(9) V. Perspectives: Inducible Systems 104(4) References 108(9)
8. Vectors and Techniques for Ectopic Gene Expression in Zebrafish 117(11) Tana M. Hyatt Stephen C. Ekker I. Introduction 117(1) II. Misexpression from DNA 118(1) III. Misexpression from RNA 118(7) References 125(3)
9. Analysis of Zebrafish Development Using Explant Culture Assays 128(31) Yevgenya Grinblat Mary Ellen Lane Charles Sagerstrom Hazel Sive I. Introduction 128(3) II. Zebrafish Explants: General Considerations 131(2) III. Materials Required 133(7) IV. Guide to Explant Isolation and Culture 140(10) V. Using Explants to Assay Induction 150(3) VI. Illustrations of Specification and Induction Assays 153(2) VII. Future Directions 155(1) References 155(4) PART IV Early Embryonic Development 159(94)
10. Embryonic Axis Formation in the Zebrafish 159(21) Mary C. Mullins I. The First Signs of Dorsal 159(1) II. Establishment of Dorsal-Ventral Polarity in Blastula Stages 160(1) III. Dorsal-Ventral Patterning 161(6) IV. Development of Midline Mesodermal Tissues 167(7) References 174(6)
11. Confocal Microscopic Analysis of Morphogenetic Movements 180(26) Mark S. Cooper Leonard A. DAmico Clarissa A. Henry I. Introduction 180(1) II. Confocal Imaging of Embryos 181(1) III. General Principles of Vital Staining 181(11) IV. Mounting Embryos for Imaging 192(1) V. Imaging Procedures 193(2) VI. Multilevel Time-Lapse Confocal Analysis 195(5) VII. Distribution of Visual Information 200(1) VIII. Confocal Imaging of Embryos Expressing Green Fluorescent Protein (GFP) 201(1) IX. Summary 201(1) References 202(4)
12. Cytoskeletal Dynamics of the Zebrafish Embryo 206(21) Jacek Topezewski Lilianna Solnica-Krezel I. Introduction 206(1) II. Cytoskeleton of the Unfertilized Egg 206(3) III. Organization and Function of the Cytoskeleton in the Zygote 209(3) IV. Cleavage and Blastula Period 212(3) V. Yolk Cell Microtubules during Epiboly 215(3) VI. Tubulin Dynamics in Neuronal Axons of Living Zebrafish Embryos 218(1) VII. Methods 219(5) References 224(3)
13. Kinesin-like Microtubule Motors in Early Development 227(26) Ming-Chyuan Chen H. William Detrich, III I. Introduction 227(2) II. Reverse Transcription-Polymerase Chain Reaction (RT-PCR) Cloning Strategy for Zebrafish Kinesin-like Protein Genes (zKLPs) 229(6) III. Production of Anti-zKLP Antibodies 235(7) IV. Expression of GFP-Tagged zKLPs in Cultured Cells 242(5) V. General Considerations 247(1) References 248(5) PART V Organogenesis 253(134)
14. Techniques in Neural Development 253(20) Cecilia B. Moens Andreas Fritz I. Introduction 253(1) II. Organizers of Anterior-Posterior Pattern in the Neural Plate 254(5) III. Zebrafish Mutations That Disrupt A-P Pattern in the Neural Tube 259(3) IV. Techniques for the Study of Neural Patterning Mutants 262(7) Appendix: Codetection of Lineage Marker and Gene Expression in Whole-Mount Embryos 269(1) References 270(3)
15. Development of the Retina 273(29) Jarema Malicki I. Introduction 273(2) II. Development of the Zebrafish Retina 275(3) III. Embryological Analysis of Wild-Type and Mutant Zebrafish Visual Systems 278(7) IV. Genetic Analysis of the Zebrafish Retina 285(9) V. Summary 294(1) References 295(7)
16. Growth Control in the Ontogenetic and Regenerating Zebrafish Fin 302(12) Stephen L. Johnson Paul Bennett I. Introduction 302(2) II. Mutations Affect the Growth, Development, and Regeneration of Fins 304(3) III. Developmental Checkpoints in Fin Growth 307(1) IV. Cellular Basis of Growth in the Fin 308(2) V. Conclusion 310(1) References 310(4)
17. Vascular and Blood Gene Expression 314(23) Leon H. Parker Leonard I. Zon Didier Y.R. Stainier I. Introduction 314(1) II. Embryonic Vascularization 314(4) III. Embryonic Hematopoiesis 318(2) IV. Vascular and Hematopoietic Gene Expression during Development 320(7) V. Mutations Affecting Vascularization and Hematopoiesis 327(5) VI. Conclusion and Future Directions 332(1) References 333(4)
18. Analysis of Hemostasis in the Zebrafish 337(22) Pudur Jagadeeswaran Yuan C. Liu John P. Sheehan I. Introduction 337(6) II. Methods 343(9) III. Future Assay Development and Applications 352(3) References 355(4)
19. Cell Lineage Tracing in Heart Development 359(8) Fabrizio C. Serluca Mark C. Fishman I. Introduction 359(1) II. Methodology 360(2) III. Analysis 362(2) IV. Summary 364(1) References 364(3)
20. Neurogenesis in Zebrafish Embryos 367(20) Ajay B. Chitnis Igor B. Dawid I. The System 367(1) II. Early Zebrafish Embryos Have a Simple Nervous System 368(2) III. Origins and Morphogenesis of the Nervous System 370(1) IV. Antagonists of Bone Morphogenetic Protein (BMP) Activity Lead to a Neural Fate in the Ectoderm 371(1) V. Fibroblast Growth Factor-8 (FGF-8) Defines Dorsoventral Positional Information in the Ectoderm 372(1) VI. Interactions with the Germ-Ring Provide Posteriorizing Signals 373(1) VII. Ablation of Cells in the Anterior Ectoderm Identifies a Novel Organizer 373(1) VIII. Patterning of the Neural Plate by Midline Signals: The Role of Factors in the Hedgehog, EGF, and TGF-Beta Families 373(2) IX. BMPs Induce Dorsal Neurons 375(1) X. Controlling the Number of Neurons: Proneuronal Domains and Proneural and Neurogenic Genes 375(3) XI. Position with Respect to Somite Boundaries Determines Motorneuron Subtype 378(2) XII. Patterning Neurogenesis in the Epiphysial Neurons 380(1) XIII. valentino (val) Suggests That Rhombomere Organization Is Preceded by the Formation of Protosegments 381(1) XIV. Perspectives 382(1) References 383(4) Index 387
Daugiau apie elektronines knygas