Avian Embryology (eBook)

Cover 
1 
Avian Embryology 
4 
Copyright Page 5
TOC$Contents 
6 
Contributors 12
Preface 16
CH$Chapter 1: Operations on Primitive Streak Stage Avian Embryos 20
I. Introduction 20
II. Staging of Embryos 21
III. Anatomy of the Primitive Streak Stage Embryo 23
IV. New Culture 25
V. Hensen's Node Grafts and Other Induction Assays 27
VI. Other Types of Graft 31
VII. Fixing Operated Embryos for Analysis 32
References 33
Part I: Embryological Microsurgery and Tissue Culture Methods 18
CH$Chapter 2: Quail-Chick Transplantations 36
I. Introduction 37
II. Differential Diagnosis of Quail and Chick Cells 40
III. Material and Equipment 44
IV. Egg Holders 45
V. Preparation and Sealing of Eggs 48
VI. Neural Tissue Transplantations 48
VII. Early Transplantations in Blastodiscs 56
VIII. Transplantation of Epiblast or Primitive Streak Fragments 57
IX. Hemopoietic Organ Rudiment Transplantations 57
X. Genetic Manipulation by Electroporation in the Quail-Chick System 64
XI. Results, Discussion, and Perspectives 66
Acknowledgments 69
References 69
CH$Chapter 3: Other Chimeras: Quail-Duck and Mouse-Chick 76
I. Introduction 77
II. Preparation of Quail, Duck, Chick, and Mouse Embryos 79
III. Generation and Analysis of Quail-Duck Chimeras 82
IV. Generation and Analysis of Mouse-Chick Chimeras 84
V. Conclusion 88
References 88
CH$Chapter 4: Manipulations of Neural Crest Cells or Their Migratory Pathways 92
I. Introduction 93
II. Preparation of Avian Neural Crest Cultures 93
III. Induction and Specification Assays for Neural Crest Cells 101
IV. Microinjection of Cells and Antibodies into Embryos 105
V. Labeling of Neural Crest Cells In Vivo with Vital Dyes 107
VI. Grafting Techniques 109
VII. Conclusions 111
References 112
CH$Chapter 5: Embryo Slices and Strips: Guidance and Adhesion Assays in the Avian Embryo 114
I. Introduction 115
II. Preparing and Culturing Embryo Slices 115
III. Preparing and Culturing Somite Strips 121
IV. Perspectives 125
References 129
CH$Chapter 6: Neural Crest, Sensory Neuron, and Muscle Cultures 132
I. Introduction 133
II. Materials 134
III. Methods 136
IV. Neural Crest Culture 137
V. Sensory Neuron Culture 139
VI. Pectoral Muscle Culture 143
VII. Results and Discussions 144
References 147
CH$Chapter 7: Methods in Avian Embryology Experimental and Molecular Manipulation of the Embryonic Chick Limb 152
I. Introduction 153
II. Visualization of the Limb Bud and Analysis of the Experimental Outcome 154
III. Proximal-Distal Limb Development 156
IV. Anterior-Posterior Patterning 164
V. Dorsal-Ventral (D-V) Patterning 167
References 167
CH$Chapter 8: Cell Division, Differentiation, and Death in Avian Embryos 170
I. Introduction 170
II. Studies in Cell Death 171
III. Studies in Cell Proliferation and Differentiation 172
IV. Selected Protocols 173
References 181
Part II: Labeling and Transgenesis Approaches 184
CH$Chapter 9: In Situ Hybridization Analysis of Chick Embryos in Whole-Mount and Tissue Sections 186
I. Introduction 187
II. Solutions 188
III. Single and Multiple Detection of RNA in Floating Sections or Whole-Mount Embryos 189
IV. Photography and Sectioning 196
V. Whole-Mount Fluorescent In Situ Hybridization 196
VI. Photography and Sectioning 199
VII. In Situ Hybridization to Tissue Sections 199
References 202
CH$Chapter 10: Vital Labeling of Embryonic Cells Using Fluorescent Dyes and Proteins 204
I. Introduction 205
II. Iontophoretic Microinjection of Lineage Tracers 206
III. Iontophoretic Application of DiI 215
IV. Relative Advantages of Dextran and DiI 217
V. Photoactivation of Fluorescent Proteins in Single Cells 217
VI. Conclusions and Emerging Technologies 225
References 225
CH$Chapter 11: Time-Lapse Imaging of the Early Avian Embryo 228
I. Introduction 229
II. The Technology of Time-Lapse Imaging 231
III. Considerations Before Time-Lapse Imaging 235
IV. Methods 239
References 250
CH$Chapter 12: Gain- and Loss-of-Function Approaches in the Chick Embryo 254
I. Gain-of-Function by Electroporation 256
II. Loss-of-Function by Electroporation 260
III. Retrovirus-Mediated Protein Expression 268
References 271
Chapter 13: Manipulation and Electroporation of CH$the Avian Segmental Plate and Somites In Vitro 274
I. Introduction 275
II. Rationale 276
III. Methods 277
IV. Materials 283
V. Discussion 284
VI. Summary 286
Acknowledgments 286
References 286
CH$Chapter 14: Transposon-Mediated Stable Integration and Tetracycline-Inducible Expression of Electroporated Transgenes in Chicken Embryos 288
I. Introduction 289
II. Tet-Inducible Expression of Electroporated Transgenes 289
III. Stable Integration of Electroporated Transgenes 293
IV. Stage-Specific Manipulation of Stably Integrated Transgenes 295
V. Summary 295
Acknowledgments 297
References 297
CH$Chapter 15: Generating Transgenic Quail using Lentiviruses 298
I. Introduction 299
II. Experimental Procedures 301
III. Concluding Remarks 308
Acknowledgments 308
References 308
Part III: Functional Genomics 312
CH$Chapter 16: Gene Discovery: Macroarrays and Microarrays 314
I. Introduction 315
II. Chicken as a Model System: Integrating Embryology and Genomics 315
III. The Array: Choosing a Platform 316
IV. Strategy: Devising the Best Screen Possible 319
V. Analysis: Sorting through Several Thousand Data Points 320
VI. Protocol: Macroarray Screening 321
References 328
CH$Chapter 17: Dissection of Chick Genomic Regulatory Regions 330
I. Introduction 331
II. Rationale 332
III. Methods 336
IV. Discussion 346
Acknowledgments 352
References 352
CH$Chapter 18: Computational Approaches to Finding and Analyzing cis-Regulatory Elements 354
I. Structure and Function of cis-Regulatory Elements 355
II. Effective cis-Regulatory Sequence Analysis 356
III. Approaches and Tools for Finding Conserved Sequence Elements 359
IV. Identifying Transcription Factor Binding Sites Computationally 369
Acknowledgments 380
References 380
CH$Chapter 19: Investigating Regulatory Factors and Their DNA Binding Affinities Through Real Time Quantitative PCR (RT-QPCR) and Chromatin Immunoprecipitation (ChIP) Assays 384
I. Introduction 385
II. Materials for RT-QPCR 390
III. General Principles and Definitions 392
IV. Types of Assays 393
V. Methods of Analysis 395
VI. Assay Setup 397
VII. ChIP and RT-QPCR 398
VIII. Conclusions and Perspectives 404
Acknowledgements 406
References 406
Relevant Websites 406
IDX$Index 408
Volumes in Series 418