Preface xi
Contributors xiii
Acknowledgments xvii
1. The Reproductive Biology and Spawning of Zebrafish in Laboratory Settings 1
1.1 Introduction 1
1.2 Overview of Zebrafish Reproductive Biology and Behavior 2
1.3 Spawning Techniques and Technology 5
1.4 Determining Factors for Reproduction in Laboratory Stocks of Zebrafish 8
1.5 Conclusions 11
References 11
2. Developmental Toxicity Assessment in Zebrafish 15
2.1 Introduction 15
2.2 Methods 16
2.3 Results 20
2.4 Discussion 23
References 25
3. Use of Emerging Models for Developmental Toxicity Testing 27
3.1 Importance of Assessing Developmental Toxicity 27
3.2 Current Methods for Assessing Developmental Toxicity 27
3.3 Use of Emerging Models for Developmental Toxicity Testing 28
3.4 New Guidelines for Chemical Testing Using Zebrafish 29
3.5 Conclusions 42
References 43
4. Assessment of Drug-Induced Cardiotoxicity in Zebrafish 45
4.1 Introduction 45
4.2 Zebrafish Heart 45
4.3 Summary of Cardiotoxicity Study Design and Results 47
4.4 Materials and Methods 47
4.5 Results 50
4.6 Conclusions 54
References 54
5. Cardiotoxicity Studies in Zebrafish 55
5.1 Introduction 55
5.2 Repolarization Toxicity 56
5.3 Initial Screening: Bradycardia 56
5.4 High-Resolution Assays of Repolarization 59
5.5 Future Directions 61
References 62
6. In Vivo Recording of the Adult Zebrafish Electrocardiogram 65
6.1 Introduction 65
6.2 Optimization of Zebrafish Electrocardiogram Recording 66
6.3 Basic Intervals 68
6.4 Drug Effects 69
6.5 Conclusions 71
References 72
7. Hematopoietic and Vascular System Toxicity 73
7.1 Introduction 73
7.2 Hematopoiesis and Vascular Development in the Zebrafish 73
7.3 Morphological and Functional Assays to Assess Toxicity 77
7.4 Summary 85
Acknowledgment 85
References 85
8. Hepatotoxicity Testing in Larval Zebrafish 89
8.1 Introduction: The Larval Zebrafish Model 89
8.2 Liver Development 89
8.3 Hepatic Gene Knockdown and Mutation 91
8.4 Hepatotoxicity Testing in Drug Discovery 92
8.5 Phenotypic-Based Larval Zebrafish Hepatotoxicity Screens 93
8.6 Secondary and Mechanistic Liver Assays 96
8.7 Conclusions 99
References 99
9. Whole Zebrafish Cytochrome P450 Assay for Assessing Drug Metabolism and Safety 103
9.1 Introduction 103
9.2 Background and Significance 104
9.3 Materials and Methods 105
9.4 Results 107
9.5 Conclusions 113
Acknowledgment 113
References 113
10. Methods for Assessing Neurotoxicity in Zebrafish 117
10.1 Introduction 117
10.2 Limitations of Current Neurotoxicity Testing 118
10.3 Assessing Neurotoxicity in Zebrafish 118
10.4 Summary 130
Acknowledgments 131
References 131
11. Zebrafish: A Predictive Model for Assessing Cancer Drug-Induced Organ Toxicity 135
11.1 Introduction 135
11.2 Materials and Methods 136
11.3 Results 139
11.4 Conclusions 149
Reference 149
12. Locomotion and Behavioral Toxicity in Larval Zebrafish: Background, Methods, and Data 151
12.1 Introduction 151
12.2 Background 152
12.3 Locomotion 153
12.4 Zebrafish Models 154
12.5 Analyzing Larval Locomotion 155
12.6 Chemical Effects on Larval Locomotion 158
12.7 Conclusions 161
Acknowledgments 162
References 162
13. Zebrafish: A Predictive Model for Assessing Seizure Liability 165
13.1 Introduction 165
13.2 Materials and Methods 167
13.3 Results 171
13.4 Conclusions 174
References 175
14. Zebrafish: A New In Vivo Model for Identifying P-Glycoprotein Efflux Modulators 177
14.1 Introduction 177
14.2 Materials and Methods 179
14.3 Results 182
14.4 Conclusions 188
Acknowledgment 188
References 189
15. Assessment of Effects on Visual Function in Larval Zebrafish 191
15.1 Introduction 191
15.2 Development of Visual System in Zebrafish 192
15.3 Methods for Assessing Visual Function in Larval Zebrafish 194
15.4 Conclusions 202
References 202
16. Development of a Hypoxia-Induced Zebrafish Choroidal Neovascularization Model 205
16.1 Introduction 205
16.2 Materials and Methods 207
16.3 Results 210
16.4 Discussion 215
Acknowledgments 217
References 217
17. Zebrafish Xenotransplant Cancer Model for Drug Screening 219
17.1 Introduction 219
17.2 Background and Significance 219
17.3 Materials and Methods 221
17.4 Results 226
17.5 Conclusions 230
References 231
18. Zebrafish Assays for Identifying Potential Muscular Dystrophy Drug Candidates 233
18.1 Introduction 233
18.2 Materials and Methods 238
18.3 Results 242
18.4 Discussion 251
Acknowledgment 252
References 252
19. Cytoprotective Activities of Water-Soluble Fullerenes in Zebrafish Models 257
19.1 Introduction 257
19.2 Materials and Methods 261
19.3 Results 265
19.4 Discussion 277
19.5 Conclusions 278
Acknowledgments 278
References 279
20. Fishing to Design Inherently Safer Nanoparticles 283
20.1 Introduction 283
20.2 Application of Embryonic Zebrafish 284
20.3 Tier 1: Rapid Toxicity Screening 286
20.4 Tier 2: Cellular Toxicity and Distribution 288
20.5 Tier 3: Molecular Expression 289
20.6 Embryonic Zebrafish Data to Design "Safer" Nanoparticles 291
20.7 Conclusions 292
References 292
21. Radiation-Induced Toxicity and Radiation Response Modifiers in Zebrafish 295
21.1 Introduction 295
21.2 Materials and Methods 296
21.3 Validation of Zebrafish Embryos as a Model System for Radiation Protectors/Sensitizers 298
21.4 Gross Morphological Alterations Associated with Radiation Exposure 299
21.5 Radiation-Associated Apoptosis Incidence 299
21.6 Radiation-Associated Gastrointestinal Toxicity 300
21.7 Radiation-Associated Nephrotoxicity 301
21.8 Ototoxicity in Irradiated Zebrafish 301
21.9 Radiation Protectors in Zebrafish 302
21.10 Summary 303
References 304
22. Caudal Fin Regeneration in Zebrafish 307
22.1 Introduction 307
22.2 Signaling and Epimorphic Regeneration 308
22.3 Caudal Fin Architecture 308
22.4 Stages of Epimorphic Regeneration 309
22.5 Methodology 311
22.6 Strategies Used to Manipulate Gene Function During Fin Regeneration 312
22.7 The Larval Fin Regeneration Model 316
22.8 Summary 317
Acknowledgments 317
References 317
Index 321
