Molecular Methods in Developmental Biology

Xenopus and Zebrafish
1. Auflage. Softcover version of original hardcover Auflage 1999
ISBN: 978-1-61737-189-9
Verlag: Humana Press

Buch, Englisch, Band 127, 217 Seiten, Paperback, Format (B × H): 155 mm x 235 mm, Gewicht: 401 g

Reihe: Methods in Molecular Biology

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.

Zielgruppe

Research

Autoren/Hrsg.

Guille, Matt

Fachgebiete

• Naturwissenschaften Biowissenschaften Entwicklungsbiologie
• Naturwissenschaften Biowissenschaften Biowissenschaften Evolutionsbiologie
• Naturwissenschaften Biowissenschaften Molekularbiologie

Inhaltsverzeichnis

The Animal Cap Assay.- Cell and Tissue Transplantation in Zebrafish Embryos.- Ribonuclease Protection Analysis of Gene Expression in Xenopus.- Quantitative Analysis of mRNA Levels in Xenopus Embryos by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR).- Wholemount In Situ Hybridization of Xenopus and Zebrafish Embryos.- In Situ Hybridization to Sections of Xenopus Embryos.- Zebrafish Immunohistochemistry.- Immunohistochemistry of Xenopus Embryos.- Preparation and Testing of Synthetic mRNA for Microinjection.- Microinjection into Xenopus Oocytes and Embryos.- Microinjection into Zebrafish Embryos.- Expression from DNA Injected into Xenopus Embryos.- Promoter Analysis in Zebrafish Embryos.- Transient Transgenesis in Xenopus laevis Facilitated by AAV-ITRs.- Band-Shift Analysis Using Crude Oocyte and Embryo Extracts from Xenopus laevis.- DNA-Footprinting using Crude Embryonic Extracts from Xenopus laevis.- Mapping Protein-DNA Interactions Using In Vivo Footprinting.