Methods in Avian Embryology (eBook)

Cover 1
Contents 6
Chapter 1. Culture and Microsurgical Manipulation of the Early Avian Embryo 16
I. General Introduction 16
II. Culture of the Avian Embryo 17
III. Tools for Microsurgery 28
IV. Microsurgical Approaches to the Young Embryo 29
References 34
Chapter 2. Quail-Chick Transplantations 38
I. Introduction 39
II. Differential Diagnosis of Quail and Chick Cells 42
III. Material and Equipment 44
IV. Preparation and Sealing of Eggs 50
V. Neural Tissue Transplantations 50
VI. Early Transplantations in Blastodiscs 55
VII. Hemopoietic Organ Rudiment Transplantations 59
VIII. Results, Discussion, and Perspectives 66
References 69
Chapter 3. Manipulations of Neural Crest Cells or Their Migratory Pathways 76
I. Introduction 76
II. Preparation of Avian Neural Crest Cultures 77
III. Microinjection of Cells and Antibodies into Embryos 85
IV. Labeling of Neural Crest Cells in Vivo with Vital Dyes 87
V. Grafting Techniques 89
VI. Conclusions 93
References 93
Chapter 4. Manipulation of the Avian Segmental Plate in Vivo 96
I. Introduction 96
II. Materials 97
III. Methods 99
IV. Critical Aspects of the Procedure 105
V. Results and Discussion 106
VI. Conclusions and Perspectives 106
References 107
Chapter 5. Somite Strips: An Embryo Fillet Preparation 108
I. Introduction 109
II. Fillet Preparation 109
III. Critical Aspects 116
IV. Analyzing Neurites to Assess Guidance Interactions 119
V. Conclusions and Perspectives 120
References 122
Chapter 6. Embryo Slices 124
I. Introduction 124
II. Cutting and Culturing Embryo Slices 125
III. Embryo Slice Characteristics 131
IV. Experimental Results Using Embryo Slices 132
V. Conclusions and Perspectives 137
References 138
Chapter 7. Operations on Limb Buds of Avian Embryos 140
I. Introduction 140
II. Preparing to Operate 141
III. Extraembryonic Membranes 146
IV. Making Various Kinds of Grafts 148
V. Manipulations Involving the Apical Ectodermal Ridge 154
VI. Grafts to Test Polarizing Activity 156
VII. Enzymatic Dissociation and Recombination of Limb Tissues 157
References 159
Chapter 8. Iontophoretic Dye Labeling of Embryonic Cells 168
I. Introduction 168
II. Iontophoretic Microinjection of Lineage Tracers 170
III. Iontophoretic Application of DiI 179
IV. Relative Advantages of Dextran and DiI 180
References 181
Chapter 9. Gene Transfer Using Replication-Defective Retroviral and Adenoviral Vectors 182
I. Introduction 183
II. Retroviral Vectors 183
III. Adenoviral Vectors 189
IV. Injection of Virus into Chick Embryos 194
V. Histology and Histochemistry 195
VI. Uses of Viral Vectors 198
References 202
Chapter 10. Manipulating Gene Expression with Replication-Competent Retroviruses 206
I. Introduction 207
II. Materials 216
III. Methods 217
IV. Results and Discussion 235
V. Conclusions and Perspectives 237
References 238
Chapter 11. In Situ Hybridization Analysis of Chick Embryos in Whole Mount and Tissue Sections 240
I. Introduction 241
II. When to Hybridize to Sections or Whole Mounts 242
III. Solutions 242
IV. Whole Mount in Situ Hybridization 243
V. Double Detection of Two RNAs 249
VI. Double Detection of RNA and Protein 250
VII. Combined DiI Labeling and in Situ Hybridization 251
VIII. In Situ Hybridization to Tissue Sections 251
IX. Troubleshooting Guide 254
References 256
Chapter 12. Micromass Cultures of Limb and Other Mesenchyme 258
I. Introduction 258
II. Micromass Culture Technique 259
III. Use of Micromass Cultures in Teratology 266
IV. Discussion and Perspectives 267
References 267
Chapter 13. Autonomic and Sensory Neuron Cultures 270
I. Introduction 270
II. Materials 272
III. Methods 273
IV. Critical Aspects of the Procedures 280
V. Results and Discussion 282
VI. Summary and Conclusions 283
References 283
Chapter 14. Retinal Cultures 286
I. Introduction 286
II. Obtaining Primary Cells 287
III. Cell Culture 296
IV. Neuronal Cell Assays 299
V. Concluding Remarks 303
References 303
Chapter 15. Migration and Adhesion Assays 306
I. Introduction 306
II. Materials 306
III. Assay Methods 308
IV. Conclusions 320
References 320
Chapter 16. Cell Division and Differentiation in Avian Embryos: Techniques for Study of Early Neurogenesis and Myogenesis 322
I. Cell Cycle and Cell Specialization 322
II. Cell Division and Neural Differentiation 324
III. Cell Division and Somite Differentiation 328
IV. Primary Methods Selected to Detect Dividing Cells 330
V. Selected Histological Procedures 337
References 348
Chapter 17. Time-Lapse Cinephotomicrogaphy, Videogaphy, and Videomicrography of the Avian Blastoderm 352
I. Introduction 352
II. General Methods 353
III. Time-Lapse Cinephotomatography, Videography, and Videomicrography Installation 362
IV. Critical Aspects of Time-Lapse Video Registration 364
V. Results and Discussion 364
VI. Conclusions and Perspectives 374
References 374
Index 376