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Methods in Molecular Biology: Molecular Embryology: Methods and Protocols
by Paul T. Sharpe
ISBN13: 9780896033870
ISBN10: 0896033872
Synopses & Reviews
Synopses & Reviews
A comprehensive guide to cutting-edge methods used today across the dramatically growing field of molecular embryology. These powerful techniques take advantage of the most commonly used vertebrate experimental models: murine embryos for their genetics, chick embryos for in vivo manipulation, zebrafish for mutagenesis, amphibian embryos, and nonvertebrate chordates. The book emphasizes the most important methods of experimental molecular biology and the particular advantages of each different species. Detailed, easy-to-follow instructions, together with relevant background information and helpful tips, ensure easy reproducibility.
Table of Contents
Part I. Mouse Embryo
The Mouse as a Developmental Model
Paul T. Sharpe
Culture of Postimplantation Mouse Embryos
Paul Martin and David L. Cockroft
Organ Culture in the Analysis of Tissue Interactions
Irma Thesleff and Carin Sahlberg
Treatment of Mice with Retinoids In Vivo and In Vitro: Skeletal Staining
Gillian M. Morriss-Kay
Cell Grafting and Labeling in Postimplantation Mouse Embryos
Gabriel A. Quinlan, Paul A. Trainor, and Patrick P. L. Tam
Production of Transgenic Rodents by the Microinjection of Cloned DNA Into Fertilized One-Celled Eggs
San Ling Si-Hoe and David Murphy
Cre Recombinase Mediated Alterations of the Mouse Genome Using Embryonic Stem Cells
Anna-Katerina Hadjantonakis, Melinda Pirity, and András Nagy
Gene Trapping in Mouse Embryonic Stem Cells
Jane Brennan and William C. Skarnes
Production and Growth of Conditionally Immortal Cell Lines from the H-2KbtsA58 Transgenic Mouse
Mark Noble
Reporter Genes for the Study of Transcriptional Regulation in Transgenic Mouse Embryos
Jonathan D. Gilthorpe and Peter W. J. Rigby
Application of lacZ Transgenic Mice to Cell Lineage Studies
Paul A. Trainor, Sheila X. Zhou, Maala Parameswaran, Gabriel A. Quinlan, Monica Gordon, Karin Sturm, and Patrick P. L. Tam
Mouse Primordial Germ Cells: Isolation and In Vitro Culture
Patricia A. Labosky and Brigid L. M. Hogan
Part II. Chicken Embryo
The Avian Embryo: An Overview
Ivor Mason
Chick Embryos: Incubation and Isolation
Ivor Mason
New Culture
Amata Hornbruch
Grafting Hensen's Node
Claudio D. Stern
Grafting of Somites
Claudio D. Stern
Manipulating the Chick Embryo: Notochord Grafts
Andrew Lumsden and Susanne Dietrich
Transplantation of Avian Neural Tissue
Sarah Guthrie
Grafting of Apical Ridge and Polarizing Region
Cheryl Tickle
Tissue Recombinations in Collagen Gels
Marysia Placzek and Kim Dale
Quail–Chick Chimeras
Marie-Aimée Teillet, Catherine Ziller, and Nicole M. Le Douarin
Using Fluorescent Dyes for Fate Mapping, Lineage Analysis, and Axon Tracing in the Chick Embryo
Jonathan D. W. Clarke
Part III. Amphibian Embryos
An Overview of Xenopus Development
C. Michael Jones and Jim Smith
Mesoderm Induction Assays
C. Michael Jones and Jim Smith
Experimental Embryological Methods for Analysis of Neural Induction in the Amphibian
Ray Keller, Ann Poznanski, and Tamira Elul
A Method for Generating Transgenic Frog Embryos
Enrique Amaya and Kristen L. Kroll
Axolotl/newt
Malcolm Maden
Part IV. Zebrafish
The Zebrafish: An Overview of Its Early Development
Nigel Holder and Qiling Xu
Small-Scale Marker-Based Screening For Mutations in Zebrafish Development
Peter D. Currie, Thomas F. Schilling, and Philip W. Ingham
Transgenic Zebrafish
Trevor Jowett
Microinjection of DNA, RNA, and Protein into the Fertilized Zebrafish Egg For Analysis of Gene Function
Nigel Holder and Qiling Xu
Retinoids in Nonmammalian Embryos
Malcolm Maden
Part V. Nonvertebrate Chordates
Protochordates
Peter W. H. Holland and Hiroshi Wada
Part VI. Retrovirus
Gene Transfer to the Rodent Embryo by Retroviral Vectors
Grace K. Pavlath and Marla B. Luskin
Gene Transfer in Avian Embryos Using Replication-Competent Retroviruses
Cairine Logan and Philippa Francis-West
Part VII. Molecular Techniques
Subtractive Hybridization and Construction of cDNA Libraries
Bruce Blumberg and Juan Carlos Izpisúa Belmonte
Differential Display of Eukaryotic mRNA
Antonio Tugores and Juan Carlos Izpísua Belmonte
RT-PCR on Embryos Using Degenerate Oligonucleotide Primers
Anthony Graham
Single-Cell RT-PCR cDNA Subtraction
Damian L. Weaver, César Núñez, Clare Brunet, Victoria Bostock, and Gerard Brady
In Situ Hybridization of Radioactive Riboprobes to RNA in Tissue Sections
Radma Mahmood and Ivor Mason. In Situ Hybridization to RNA in Whole Embryos
Huma Shamim, Radma Mahmood, and Ivor Mason
Wholemount In Situ Hybridization to Xenopus Embryos
C. Michael Jones and James C. Smith
Wholemount In Situ Hybridization to Amphioxus Embryo
Peter W. H. Holland
In Situ Hybridization to Sections (Nonradioactive)
Maria Rex and Paul J. Scotting
Immunohistochemistry Using Polyester Wax
Andrew Kent
Immunohistochemistry on Whole Embryos
Ivor Mason
Whole Embryo Assays for Programmed Cell Death
Anthony Graham
Gene Interference Using Antisense Oligodeoxynucleotides on Whole Chick Embryos: Optimal Ring and Roller-Bottle Culture Technique
Jonathan Cooke and Alison Isaac
Protein Techniques: Immunoprecipitation, In Vitro Kinase Assays, and Western Blotting
David I. Jackson and Clive Dickson
Part VIII. Microscopy and Photography
Microscopy and Photomicrography Techniques
Richard J. T. Wingate
Index
Part I. Mouse Embryo
The Mouse as a Developmental Model
Paul T. Sharpe
Culture of Postimplantation Mouse Embryos
Paul Martin and David L. Cockroft
Organ Culture in the Analysis of Tissue Interactions
Irma Thesleff and Carin Sahlberg
Treatment of Mice with Retinoids In Vivo and In Vitro: Skeletal Staining
Gillian M. Morriss-Kay
Cell Grafting and Labeling in Postimplantation Mouse Embryos
Gabriel A. Quinlan, Paul A. Trainor, and Patrick P. L. Tam
Production of Transgenic Rodents by the Microinjection of Cloned DNA Into Fertilized One-Celled Eggs
San Ling Si-Hoe and David Murphy
Cre Recombinase Mediated Alterations of the Mouse Genome Using Embryonic Stem Cells
Anna-Katerina Hadjantonakis, Melinda Pirity, and András Nagy
Gene Trapping in Mouse Embryonic Stem Cells
Jane Brennan and William C. Skarnes
Production and Growth of Conditionally Immortal Cell Lines from the H-2KbtsA58 Transgenic Mouse
Mark Noble
Reporter Genes for the Study of Transcriptional Regulation in Transgenic Mouse Embryos
Jonathan D. Gilthorpe and Peter W. J. Rigby
Application of lacZ Transgenic Mice to Cell Lineage Studies
Paul A. Trainor, Sheila X. Zhou, Maala Parameswaran, Gabriel A. Quinlan, Monica Gordon, Karin Sturm, and Patrick P. L. Tam
Mouse Primordial Germ Cells: Isolation and In Vitro Culture
Patricia A. Labosky and Brigid L. M. Hogan
Part II. Chicken Embryo
The Avian Embryo: An Overview
Ivor Mason
Chick Embryos: Incubation and Isolation
Ivor Mason
New Culture
Amata Hornbruch
Grafting Hensen's Node
Claudio D. Stern
Grafting of Somites
Claudio D. Stern
Manipulating the Chick Embryo: Notochord Grafts
Andrew Lumsden and Susanne Dietrich
Transplantation of Avian Neural Tissue
Sarah Guthrie
Grafting of Apical Ridge and Polarizing Region
Cheryl Tickle
Tissue Recombinations in Collagen Gels
Marysia Placzek and Kim Dale
Quail–Chick Chimeras
Marie-Aimée Teillet, Catherine Ziller, and Nicole M. Le Douarin
Using Fluorescent Dyes for Fate Mapping, Lineage Analysis, and Axon Tracing in the Chick Embryo
Jonathan D. W. Clarke
Part III. Amphibian Embryos
An Overview of Xenopus Development
C. Michael Jones and Jim Smith
Mesoderm Induction Assays
C. Michael Jones and Jim Smith
Experimental Embryological Methods for Analysis of Neural Induction in the Amphibian
Ray Keller, Ann Poznanski, and Tamira Elul
A Method for Generating Transgenic Frog Embryos
Enrique Amaya and Kristen L. Kroll
Axolotl/newt
Malcolm Maden
Part IV. Zebrafish
The Zebrafish: An Overview of Its Early Development
Nigel Holder and Qiling Xu
Small-Scale Marker-Based Screening For Mutations in Zebrafish Development
Peter D. Currie, Thomas F. Schilling, and Philip W. Ingham
Transgenic Zebrafish
Trevor Jowett
Microinjection of DNA, RNA, and Protein into the Fertilized Zebrafish Egg For Analysis of Gene Function
Nigel Holder and Qiling Xu
Retinoids in Nonmammalian Embryos
Malcolm Maden
Part V. Nonvertebrate Chordates
Protochordates
Peter W. H. Holland and Hiroshi Wada
Part VI. Retrovirus
Gene Transfer to the Rodent Embryo by Retroviral Vectors
Grace K. Pavlath and Marla B. Luskin
Gene Transfer in Avian Embryos Using Replication-Competent Retroviruses
Cairine Logan and Philippa Francis-West
Part VII. Molecular Techniques
Subtractive Hybridization and Construction of cDNA Libraries
Bruce Blumberg and Juan Carlos Izpisúa Belmonte
Differential Display of Eukaryotic mRNA
Antonio Tugores and Juan Carlos Izpísua Belmonte
RT-PCR on Embryos Using Degenerate Oligonucleotide Primers
Anthony Graham
Single-Cell RT-PCR cDNA Subtraction
Damian L. Weaver, César Núñez, Clare Brunet, Victoria Bostock, and Gerard Brady
In Situ Hybridization of Radioactive Riboprobes to RNA in Tissue Sections
Radma Mahmood and Ivor Mason. In Situ Hybridization to RNA in Whole Embryos
Huma Shamim, Radma Mahmood, and Ivor Mason
Wholemount In Situ Hybridization to Xenopus Embryos
C. Michael Jones and James C. Smith
Wholemount In Situ Hybridization to Amphioxus Embryo
Peter W. H. Holland
In Situ Hybridization to Sections (Nonradioactive)
Maria Rex and Paul J. Scotting
Immunohistochemistry Using Polyester Wax
Andrew Kent
Immunohistochemistry on Whole Embryos
Ivor Mason
Whole Embryo Assays for Programmed Cell Death
Anthony Graham
Gene Interference Using Antisense Oligodeoxynucleotides on Whole Chick Embryos: Optimal Ring and Roller-Bottle Culture Technique
Jonathan Cooke and Alison Isaac
Protein Techniques: Immunoprecipitation, In Vitro Kinase Assays, and Western Blotting
David I. Jackson and Clive Dickson
Part VIII. Microscopy and Photography
Microscopy and Photomicrography Techniques
Richard J. T. Wingate
Index
