Molecular Embryology: Methods and Protocols

Cover
Paul T. Sharpe, Ivor Mason
Springer Science & Business Media, 1999 - 756 Seiten
In Molecular Embryology, expert investigators provide a comprehensive guide to the cutting-edge methods used today across the dramatically growing field of vertebrate 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 major techniques of experimental molecular biology and the particular advantages of each different species are emphasized. Detailed, easy-to-follow protocols, together with relevant background information and helpful tips, optimize the methods for success. Molecular Embryology brings together in one volume all the major techniques and common experimental species needed to study the mechanisms of biological development in vertebrates. Bound to become a standard reference in this field, the book makes it possible for experienced and novice researchers alike to move between embryos of diverse vertebrate classes as their project progresses, ensuring their ability to utilize the experimental advantages of different systems to address specific developmental questions.
 

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Inhalt

The Mouse as a Developmental Model
4
Culture of Postimplantation Mouse Embryos
7
Organ Culture in the Analysis of Tissue Interactions
23
Treatment of Mice with Retinoids In Vivo and In Vitro Skeletal Staining
33
Cell Grafting and Labeling in Postimplantation Mouse Embryos
41
Production of Transgenic Rodents by the Microinjection of Cloned DNA into Fertilized OneCelled Eggs
61
Cre Recombinase Mediated Alterations of the Mouse Genome Using Embryonic Stem Cells
101
Gene Trapping in Mouse Embryonic Stem Cells
123
ZEBRAFISH
429
The Zebrafish An Overview of Its Early Development
431
SmallScale MarkerBased Screening for Mutations in Zebrafish Development
441
Transgenic Zebrafish
461
Microinjection of DNA RNA and Protein into the Fertilized Zebrafish Egg for Analysis of Gene Function
487
Retinoids in Nonmammalian Embryos
491
NONVERTEBRATE CHORDATES
511
Protochordates
513

Production and Growth of Conditionally Immortal Cell Lines from the H2KbtsA58 Transgenic Mouse
139
Reporter Genes for the Study of Transcriptional Regulation in Transgenic Mouse Embryos
159
Application of lacZ Transgenic Mice to Cell Lineage Studies
183
Mouse Primordial Germ Cells Isolation and In Vitro Culture
201
CHICKEN EMBRYO
213
The Avian Embryo An Overview
215
Chick Embryos Incubation and Isolation
221
New Culture
225
Grafting Hensens Node
245
Grafting of Somites
255
Notochord Grafts
265
Transplantation of Avian Neural Tissue
273
Grafting of Apical Ridge and Polarizing Region
280
Tissue Recombinations in Collagen Gels
293
QuailChick Chimeras
305
Using Fluorescent Dyes for Fate Mapping Lineage Analysis and Axon Tracing in the Chick Embryo
319
AMPHIBIAN EMBRYO
329
An Overview of Xenopus Development
331
Mesoderm Induction Assays
341
Experimental Embryological Methods for Analysis of Neural Induction in the Amphibian
351
A Method for Generating Transgenic Frog Embryos
393
Axolotlnewt
415
RETROVIRUSES
517
Gene Transfer to the Rodent Embryo by Retroviral Vectors
519
Gene Transfer in Avian Embryos Using ReplicationCompetent Retroviruses
539
MOLECULAR TECHNIQUES
553
Subtractive Hybridization and Construction of cDNA Libraries
555
Differential Display of Eukaryotic mRNA
575
RTPCR on Embryos Using Degenerate Oligonucleotide Primers
591
SingleCell RTPCR cDNA Subtraction
601
In Situ Hybridization of Radioactive Riboprobes to RNA in Tissue Sections
611
In Situ Hybridization to RNA in Whole Embryos
623
Wholemount In Situ Hybridization to Xenopus Embryos
635
Wholemount In Situ Hybridization to Amphioxus Embryos
641
In Situ Hybridization to Sections Nonradioactive
645
Immunohistochemistry Using Polyester Wax
655
Immunohistochemistry on Whole Embryos
663
Whole Embryo Assays for Programmed Cell DeatH
667
Gene Interference Using Antisense Oligodeoxynucleotides on Whole Chick Embryos Optimal Ring and RollerBottle Culture Technique
673
Protein Techniques Immunoprecipitation In Vitro Kinase Assays and Western Blotting
699
MICROSCOPY AND PHOTOGRAPHY
709
Microscopy and Photomicrography Techniques
711
Index
735
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