Molecular Methods in Developmental Biology: Xenopus and Zebrafish / Edition 1

by Matt Guille

Overview

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 immunohishemistry, and good for observing mutations in genetic screens.

Product Details

ISBN-13:	9781617371899
Publisher:	Springer-Verlag New York, LLC
Publication date:	11/10/2010
Series:	Methods in Molecular Biology , #127
Edition description:	Softcover reprint of hardcover 1st ed. 1999
Pages:	217
Product dimensions:	5.98(w) x 9.02(h) x 0.02(d)

Table of Contents

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 Immunohishemistry.- Immunohishemistry 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.

Editorial Reviews

Studies of the South African clawed frog, ( or ), and the zebrafish, , showcase the latest readily reproducible experimental techniques in the field of molecular biology intended to broaden understanding of how a fertilized egg develops into an adult. The former animal model offers large embryos that can be readily surgically manipulated, while the embryos of the latter are transparent. In 17 contributions, experts review in detail key methods in such areas as: gel shift assays, cell and tissue transplantation, quantitative and spatial analysis of mRNA and proteins, immunohistochemistry, microinjection, and in vitro and in vivo footprinting. Annotation c. Book News, Inc., Portland, OR (booknews.com)

Booknews

"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

From the Publisher