Molecular Methods in Developmental Biology: Xenopus and Zebrafish

ISBN-13: 9781617371899 / Angielski / Miękka / 2010 / 217 str.

Matt Guille

Opis

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.

Recenzje

"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

Spis treści

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.

Blurb

In Molecular Methods in Developmental Biology: Xenopus and Zebrafish, Matthew Guille assembles a hands-on collection of basic and essential molecular and embryological techniques for studying Xenopus and zebrafish. Easily reproducible and designed to succeed, these detailed methods include cellular techniques, techniques for the quantitative and spatial analysis of mRNA and proteins, and techniques for the expression of gene products in embryos. More specialized methods enable users to analyze promoters and transcription factors during early development, and include gel shift assays, as well as in vitro and in vivo footprinting. Wherever possible, these experimental approaches are applied to both Xenopus and zebrafish.
Molecular Methods in Developmental Biology: Xenopus and Zebrafish affords newcomers rapid access to a wide variety of key techniques in developmental research, and offers experienced investigators both new techniques from experts who have fine-tuned them for best results, and a plethora of time-saving tips. State-of-the-art and readily reproducible, these powerful methods constitute today's gold-standard laboratory manual for understanding the interactions responsible for development.