Molecular Methods in Developmental Biology

1. Front Matter

   Pages i-xii

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2. The Animal Cap Assay

   • Jeremy Green

   Pages 1-13

3. Cell and Tissue Transplantation in Zebrafish Embryos

   • Toshiro Mizuno, Minori Shinya, Hiroyuki Takeda

   Pages 15-28

4. Ribonuclease Protection Analysis of Gene Expression in Xenopus

   • Craig S. Newman, Paul A. Krieg

   Pages 29-40

5. Quantitative Analysis of mRNA Levels in Xenopus Embryos by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR)

   • Oliver C. Steinbach, Ralph A. W. Rupp

   Pages 41-56

6. Wholemount In Situ Hybridization of Xenopus and Zebrafish Embryos

   • Joanne Broadbent, E. Mary Read

   Pages 57-67

7. In Situ Hybridization to Sections of Xenopus Embryos

   • David Bertwistle

   Pages 69-76

8. Zebrafish Immunohistochemistry

   • Rachel Macdonald

   Pages 77-88

9. Immunohistochemistry of Xenopus Embryos

   • Carl Robinson, Matt Guille

   Pages 89-97

10. Preparation and Testing of Synthetic mRNA for Microinjection

    • Wendy Moore, Matt Guille

    Pages 99-109

11. Microinjection into Xenopus Oocytes and Embryos

    • Matt Guille

    Pages 111-123

12. Microinjection into Zebrafish Embryos

    • Qiling Xu

    Pages 125-132

13. Expression from DNA Injected into Xenopus Embryos

    • Ondine Cleaver, Paul A. Krieg

    Pages 133-153

14. Promoter Analysis in Zebrafish Embryos

    • Jos Joore

    Pages 155-166

15. Transient Transgenesis in Xenopus laevis Facilitated by AAV-ITRs

    • Yuchang Fu, Donghui Kan, Sylvia Evans

    Pages 167-173

16. Band-Shift Analysis Using Crude Oocyte and Embryo Extracts from Xenopus laevis

    • Rob Orford, Matt Guille

    Pages 175-186

17. DNA-Footprinting using Crude Embryonic Extracts from Xenopus laevis

    • Rob Orford, Darren Mernagh, Matt Guille

    Pages 187-198

18. Mapping Protein-DNA Interactions Using In Vivo Footprinting

    • David Warshawsky, Leo Miller

    Pages 199-212

19. Back Matter

    Pages 213-217

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The process whereby a single cell, the fertilized egg, develops into an adult has fascinated for centuries. Great progress in understanding that process, h- ever, has been made in the last two decades, when the techniques of molecular biology have become available to developmental biologists. By applying these techniques, the exact nature of many of the interactions responsible for forming the body pattern are now being revealed in detail. Such studies are a large, and it seems ever-expanding, part of most life-science groups. It is at newcomers to this field that this book is primarily aimed. A number of different plants and animals serve as common model org- isms for developmental studies. In Molecular Methods in Developmental Bi- ogy: Xenopus and Zebrafish, a range of the molecular methods applicable to two of these organisms are described, these are the South African clawed frog, Xenopus laevis, and the zebrafish, Brachydanio rerio. The embryos of both of these species develop rapidly and externally, making them particularly suited to investigations of early vertebrate development. However, both Xenopus and zebrafish have their own advantages and disadvantages. Xenopus have large, robust embryos that can be manipulated surgically with ease, but their pseudotetraploidy and long generation time make them unsuitable candidates for genetics. This disadvantage may soon be overcome by using the diploid Xenopus tropicalis, and early experiments are already underway. The transp- ent embryos of zebrafish render them well-suited for in situ hybridization and immunohistochemistry, and good for observing mutations in genetic screens.

"Readers will find the systematic description of methods, illustration of structures, tables and figures, along with comments on optimizing the methods for use in other systems, very useful. ...this excellent book will facilitate studies on the lipidation of proteins. It should be useful to both beginning and experienced investigators in the field." - Analytical Biochemistry

"...this compact book comprises concise and very well structured chapters on both basic and advanced molecular and embryological technique used in studies of early stages of vertebrate development on two model species, the South Africa clawed frog Xenopus loevis and the zebrafish Brachydanio rerio....All chapters have been written by authors with first-hand expertise in individual fields, mostly those in whose laboratories the methods in question have been developed, or at least optimized. Close attention is paid to meticulous description of possible pitfalls and drawbacks, and essential steps are stressed....the book contains a wealth of information, cues on methodology and their critical assessment. It is an almost indispensable guide for anybody working in the field of vertebrate development biology."-Folia Microbiologica

"...this book is a solid introduction to some of the common techniques in zebrafish and Xenopus and I think would be of good use to its intended audience of new workers in the field." - Newsletter of the British Society of Developmental Biology

• School of Biological Sciences, University of Portsmouth, Portsmouth, UK

  Matthew Guille

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