Developmental Biology Protocols, Part 2

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Rocky S. Tuan, Cecilia W. Lo
Springer Science & Business Media, 2000 - Science - 530 pages
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The molecular biology revolution has transformed developmental biology into one of the most exciting and fruitful fields in experimental biomedical research today. In Developmental Biology Protocols, established leaders in this field demonstrate this achievement with a comprehensive collection of cutting-edge protocols for studying and analyzing the events of embryonic development. Drawing on state-of-the-art cellular and molecular techniques, as well as new and sophisticated imaging and information technologies, this 2nd volume of three describes state-of-the-art methods for the study of organogenesis, the analysis of abnormal development and teratology, the screening and mapping of novel genes and mutations, and the application of transgenesis, including the production of transgenic animals and gene knockouts. The 1st and 3rd volumes in this seminal set complete today's widest-ranging collection of techniques designed to decipher the exact cellular, molecular, and genetic mechanisms that control the form, structure, and function of the developing embryo. Volume 1 presents readily reproducible methods for establishing and characterizing several widely used experimental model systems, for both the study of developmental patterns and morphogenesis, and the examination of embryo structure and function. In addition, there are step-by-step methods for the analysis of cell lineage, the production and use of chimeras, and the experimental molecular manipulation of embryos, including the application of viral vectors. No less innovative, volume 3 introduces powerful techniques for the manipulation of developmental gene expression and function, the analysis of gene expression, the characterization of tissue morphogenesis and development, the in vitro study of differentiation and development, and the genetic analysis of developmental models of diseases. Highly practical and richly annotated, the three volumes of Developmental Biology Protocols describe multiple experimental systems and details techniques adopted from the broadest array of biomedical disciplines. Every researcher will not only better understand the principles, background, and rationale for how form and function are elaborated in an organism, but also gain full practical access to today's best methods for its analysis.
 

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Contents

Developmental Biology Protocols Overview II
4
ORGANOGENESIS
6
Drosophila as a Genetic Tool to Define Vertebrate Pathway Players
7
Bioassays for Studying the Role of the Peptide Growth Factor Activin in Early Amphibian Embryogenesis
15
Analysis of Mammary Gland Morphogenesis
27
Specification of Cardiac Mesenchyme and Heart Morphogenesis In Vitro
39
Craniofacial Development and Patterning
45
Craniofacial Skeletal Morphogenesis In Vitro
55
Cardiac Morphogenesis and Dysmorphogenesis
239
Analysis of Two Aspects of LeftRight Patterning of the Vertebrate Heart Heart Tube Position and Heart Tube Chirality
261
Biologically Based Risk Assessment Models for Developmental Toxicity
271
SCREENING AND MAPPING OF NOVEL GENES AND MUTATIONS
283
Positional Cloning
285
Gene Trapping in Embryonic Stem Cells In Vitro to Identify Novel Developmentally Regulated Genes in the Mouse
297
PCRBased Cloning of Cortically Localized RNAs from Xenopus Oocytes
309
Analysis of mRNA Expression During Preimplantation Development
315

Skeletal Morphogenesis
61
Transplantation and Culture Techniques for the Analysis of Urodele Limb Regeneration
71
Retroviral Infection of TCell Precursors in Thymic Organ Culture
79
Assay for the Isolation of Hepatogenic Factors Key Molecules in Hepatocyte Formation and Liver Morphogenesis
87
Skin Morphogenesis Embryonic Chicken Skin Explant Cultures
101
Apoptosis in Development
107
Methods to Detect Patterns of Cell Death in Drosophila
115
ABNORMAL DEVELOPMENT AND TERATOLOGY
123
Mechanisms of Neurulation
125
Neurulation and Neural Tube Closure Defects
135
Neural Tube Defects
161
Experimental Manipulation and Morphometric Analysis of Neural Tube Development
167
Isolation of Neuroepithelium and Formation of Minispheres
179
Examination of Normal and Abnormal Placentation in the Mouse
185
Palatal Dysmorphogenesis Palate Organ Culture
195
Palatal Dysmorphogenesis Quantitative RTPCR
203
Transspecies Grafting as a Tool to Understand the Basis of Murine Developmental Limb Abnormalities
219
Assessment of Laterality Defects in Rodent Embryos
227
Differential Screens with Subtracted PCRGenerated cDNA Libraries from Subregions of Single Mouse Embryos
333
HPLCBased mRNA Differential Display
345
TRANSGENESIS PRODUCTION AND GENE KNOCKOUT
351
Production of Transgenic Drosophila
353
Transgenic Manipulation of the Sea Urchin Embryo
365
Transgenic Zebrafish
375
Production of Avian Chimeras and Germline Transmission
385
Incorporation of Genetically Modified Cells in Chicken Chimeras
391
LongTerm Culture of Chicken Blastodermal Cells CBCs and Selection of Transfected CBCs Using Antibiotic Resistance
399
Nuclear Transplantation and Cloning in Mammals
405
Production of Transgenic Mice
427
Production of Transgenic Mice with Yeast Artificial Chromosomes
435
GeneTargeting Strategies
455
Chimeric Animals and Germline Transmission
465
Conditional Gene Knockout Using Cre Recombinase
477
Application of CreloxP in Drosophila SiteSpecific Recombination and Transgene Coplacement
487
Index
497
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Tuan, Thomas Jefferson University, Philadelphia, PA.

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