Methods in Cell Biology Vol. 59 : The Zebrafish, Biology

by Detrich, H. William; Matsudaira, Paul

• ISBN13: 9780125441612
• ISBN10: 0125441614
• Format: Hardcover
• Copyright: 1999-01-01
• Publisher: Elsevier Science Serials

Summary

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

Table of Contents

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. D'Amico
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