Molecular Embryology: Methods & Protocols,2/e > Cell Biology

Hardcover - 400 pages . 2/e
In Molecular Embryology: Methods and Protocols, Second Edition, expert investigators provide a comprehensive guide to the cutting-edge methods used across the dramatically growing field of vertebrate molecular embryology. Time-tested 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 second edition collects classic protocols which have become standard techniques in the laboratory and presents them in a complementary fashion with novel and emerging approaches, allowing researcher to become more familiar with commonly studied embryos used in biomedical research. Insightful to the experienced professional, Molecular Embryology: Methods and Protocols, Second Edition, presents cutting-edge findings of perhaps the greatest period in growth and productivity in the field of developmental biology.

Pt. I The Mouse Embryo 1
1 The Mouse as a Developmental Model 3
2 Culture of Postimplantation Mouse Embryos 7
3 Organ Culture in the Analysis of Tissue Interactions 23
4 Treatment of Mice with Retinoids In Vivo and In Vitro: Skeletal Staining 33
5 Cell Grafting and Labeling in Postimplantation Mouse Embryos 41
6 Production of Transgenic Rodents by the Microinjection of Cloned DNA into Fertilized One-Celled Eggs 61
7 Cre Recombinase Mediated Alterations of the Mouse Genome Using Embryonic Stem Cells 101
8 Gene Trapping in Mouse Embryonic Stem Cells 123
9 Production and Growth of Conditionally Immortal Cell Lines from the H-2K[superscript b]tsA58 Transgenic Mouse 139
10 Reporter Genes for the Study of Transcriptional Regulation in Transgenic Mouse Embryos 159
11 Application of IacZ Transgenic Mice to Cell Lineage Studies 183
12 Mouse Primordial Germ Cells: Isolation and In Vitro Culture 201
Pt. II Chicken Embryo 213
13 The Avian Embryo: An Overview 215
14 Chick Embryos: Incubation and Isolation 221
15 New Culture 225
16 Grafting Hensen''s Node 245
17 Grafting of Somites 255
18 Notochord Grafts 265
19 Transplantation of Avian Neural Tissue 273
20 Grafting of Apical Ridge and Polarizing Region 281
21 Tissue Recombinations in Collagen Gels 293
22 Quail-Chick Chimeras 305
23 Using Fluorescent Dyes for Fate Mapping, Lineage Analysis, and Axon Tracing in the Chick Embryo 319
Pt. III Amphibian Embryo 329
24 An Overview of Xenopus Development 331
25 Mesoderm Induction Assays 341
26 Experimental Embryological Methods for Analysis of Neural Induction in the Amphibian 351
27 A Method for Generating Transgenic Frog Embryos 393
28 Axolotl/newt 415
Pt. IV Zebrafish 429
29 The Zebrafish: An Overview of Its Early Development 431
30 Small-Scale Marker-Based Screening for Mutations in Zebrafish Development 441
31 Transgenic Zebrafish 461
32 Microinjection of DNA, RNA, and Protein into the Fertilized Zebrafish Egg for Analysis of Gene Function 487
33 Retinoids in Nonmammalian Embryos 491
Pt. V Nonvertebrate Chordates 511
34 Protochordates 513
Pt. VI Retroviruses 517
35 Gene Transfer to the Rodent Embryo by Retroviral Vectors 519
36 Gene Transfer in Avian Embryos Using Replication-Competent Retroviruses 539
Pt. VII Molecular Techniques 553
37 Subtractive Hybridization and Construction of cDNA Libraries 555
38 Differential Display of Eukaryotic mRNA 575
39 RT-PCR on Embryos Using Degenerate Oligonucleotide Primers 591
40 Single-Cell RT-PCR cDNA Subtraction 601
41 In Situ Hybridization of Radioactive Riboprobes to RNA in Tissue Sections 611
42 In Situ Hybridization to RNA in Whole Embryos 623
43 Wholemount In Situ Hybridization to Xenopus Embryos 635
44 Whole-Mount In Situ Hybridization to Amphioxus Embryos 641
45 In Situ Hybridization to Sections (Nonradioactive) 645
46 Immunohistochemistry Using Polyester Wax 655
47 Immunohistochemistry on Whole Embryos 663
48 Whole Embryo Assays for Programmed Cell Death 667
49 Gene Interference Using Antisense Oligodeoxynucleotides on Whole Chick Embryos: Optimal Ring and Roller-Bottle Culture Technique 673
50 Protein Techniques: Immunoprecipitation, In Vitro Kinase Assays, and Western Blotting 699
Pt. VIII Microscopy and Photography 709
51 Microscopy and Photomicrography Techniques 711
Index 735

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