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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.

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.

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The Animal Cap Assay, Jeremy Green. Cell and Tissue Transplantation in Zebrafish Embryos, Toshiro Mizuno, Minori Shinya, and Hiroyuki Takeda. Ribonuclease Protection Analysis of Gene Expression in Xenopus, Craig S. Newman and Paul A. Krieg. Quantitative Analysis of mRNA Levels in Xenopus Embryos by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR), Oliver C. Steinbach and Ralph A. W. Rupp. Wholemount In Situ Hybridization of Xenopus and Zebrafish Embryos, Joanne Broadbent and E. Mary Read. In Situ Hybridization to Sections of Xenopus Embryos, David Bertwistle. Zebrafish Immunohistochemistry, Rachel Macdonald. Immunohistochemistry of Xenopus Embryos, Carl Robinson and Matthew Guille. Preparation and Testing of Synthetic mRNA for Microinjection, Wendy Moore and Matthew Guille. Microinjection into Xenopus Oocytes and Embryos, Matthew Guille. Microinjection into Zebrafish Embryos, Qiling Xu. Expression from DNA Injected into Xenopus Embryos, Ondine Cleaver and Paul A. Krieg. Promoter Analysis in Zebrafish Embryos, Jos Joore. Transient Transgenesis in Xenopus laevis Facilitated by AAV-ITRs, Yuchang Fu, Donghui Kan, and Sylvia Evans. Band-Shift Analysis Using Crude Oocyte and Embryo Extracts from Xenopus laevis, Rob Orford and Matthew Guille. DNA Footprinting Using Crude Embryo Extracts from Xenopus laevis, Rob Orford, Darren Mernagh, and Matthew Guille. Mapping Protein-DNA Interactions Using In Vivo Footprinting, David Warshawsky and Leo Miller. Index.

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