SECTION 1: HUSBANDRY Editors: Richard M. Harland and Hazel L. Sive INTRODUCTION Xenopus Husbandry Richard M. Harland and Hazel L. Sive PROTOCOLS 1. Animal Maintenance Systems: Xenopus laevis Nikko-Ideen Shaidani, Sean McNamara, Marcin Wlizla, and Marko E. Horb 2. Animal Maintenance Systems: Xenopus tropicalis Nikko-Ideen Shaidani, Sean McNamara, Marcin Wlizla, and Marko E.
Horb 3. Raising and Maintaining Xenopus tropicalis from Tadpole to Adult Maura Ann Lane, Michael Slocum, and Mustafa K. Khokha 4. Defining the Specific Pathogen-free State of Xenopus Species Using TaqMan Assays Casey L Hensley, Kaitlin M Bowes, and Sanford H. Feldman SECTION 2: EGGS TO ADULT Editors: Hazel L. Sive and Richard M. Harland INTRODUCTION Obtaining Eggs and Embryos Hazel L. Sive and Richard M.
Harland PROTOCOLS 1. Obtaining Xenopus laevis Eggs Nikko-Ideen Shaidani, Sean McNamara, Marcin Wlizla, and Marko E. Horb 2. Obtaining Xenopus laevis Embryos Nikko-Ideen Shaidani, Sean McNamara, Marcin Wlizla, and Marko E. Horb 3. How to Grow Xenopus laevis Tadpole Stages to Adult Shoko Ishibashi and Enrique Amaya 4. Obtaining Xenopus tropicalis Eggs Maura Ann Lane, Emily K Mis, and Mustafa K. Khokha 5.
Obtaining Xenopus tropicalis Embryos by In Vitro Fertilization Maura Ann Lane and Mustafa K. Khokha 6. Obtaining Xenopus tropicalis Embryos by Natural Mating Maura Lane and Mustafa K. Khokha 7. Microinjection of Xenopus tropicalis Embryos Maura Lane, Emily K. Mis, and Mustafa K. Khokha 8 Best Practices for Xenopus tropicalis Husbandry Takuya Nakayama and Robert M. Grainger SECTION 3: OOCYTES AND OOGENESIS Editor: Kimberly L.
Mowry INTRODUCTION Using the Xenopus Oocyte Toolbox Kimberly L. Mowry PROTOCOLS 1. Isolation of Xenopus Oocytes Karen Newman, Tristan Aguero, and Mary Lou King 2. Isolation and Analysis of Xenopus Germinal Vesicles Garry T. Morgan 3. Microinjection of Xenopus Oocytes Tristan Aguero, Karen Newman, and Mary Lou King 4. Oocyte Host Transfer and Maternal mRNA Depletion Experiments in Xenopus Douglas W. Houston 5.
Heterologous Protein Expression in the Xenopus Oocyte Jonathan S. Marchant 6. Patch Clamp and Perfusion Techniques to Study Ion Channels Expressed in Xenopus Oocytes Guohui Zhang and Jianmin Cui 7. Whole-Mount In Situ Hybridization of Xenopus Oocytes Diana Bauermeister and Tomas Pieler 8. Whole-Mount Immunofluorescence for Visualizing Endogenous Proteins and Injected RNAs in Xenopus Oocytes Samantha P. Jeschonek and Kimberly L. Mowry 9. Fluorescence In Situ Hybridization of Cryosectioned Xenopus Oocytes Christopher R.
Neil and Kimberly L. Mowry SECTION 4: EMBRYOLOGY I Editor: Sally A. Moody INTRODUCTION Analysis of Cell Fate Commitment in Xenopus Embryos Sally A. Moody PROTOCOLS 1. Lineage Tracing and Fate Mapping in Xenopus Embryos Sally A. Moody 2. Microinjection of Xenopus Embryos with mRNAs and Oligonucleotides Sally A. Moody 3.
Microinjection of DNA Constructs into Xenopus Embryos for Gene Misexpression and cis-Regulatory Module Analysis Yuuri Yasuoka and Masanori Taira 4. Whole-Mount In Situ Hybridization of Xenopus Embryos Jean-Pierre Saint-Jeannet 5. Whole-Mount Immunocytochemistry in Xenopus Michael W. Klymkowsky 6. Cleavage Blastomere Explant Culture in Xenopus Sally A. Moody 7. Cleavage Blastomere Deletion and Transplantation to Test Cell Fate Commitment in Xenopus Sally A. Moody 8.
Dissecting and Culturing Animal Cap Explants Kevin S. Dingwell and Jim C. Smith SECTION 5 EMBRYOLOGY II Editor: Sally A. Moody INTRODUCTION Xenopus Explants and Transplants Sally A. Moody PROTOCOLS 1. Spemann-Mangold Grafts Helene Cousin 2. Einsteck Transplants Helene Cousin 3. Microsurgical Methods to Isolate and Culture the Early Gastrula Dorsal Marginal Zone Lance A.
Davidson 4. Microsurgical Manipulations to Isolate Collectively Migrating Mesendoderm Lance A. Davidson 5. Microsurgical Methods to Make the Keller Sandwich Explant and the Dorsal Isolate Lance A. Davidson 6. Cranial Neural Crest Explants Helene Cousin and Dominique Alfandari 7. Cranial Neural Crest Transplants Helene Cousin 8. In Vitro Induction of Xenopus Embryonic Organs Using Animal Cap Cells Takashi Ariizumi, Tatsuo Michiue, and Makoto Asashima 9.
Special Considerations for Making Explants and Transplants with Xenopus tropicalis Marilyn Fisher and Robert M. Grainger SECTION 6: CELL BIOLOGY Editor: Anna Philpott INTRODUCTION The Use of Xenopus for Cell Biology Applications Anna Philpott PROTOCOLS 1. In Vivo Assessment of Neural Precursor Cell Cycle Kinetics in the Amphibian Retina Morgane Locker and Muriel Perron 2. Applying Tensile and Compressive Force to Xenopus Animal Cap Tissue Georgina K. Goddard, Nawseen Tarannum, and Sarah Woolner 3. Analysis of Phosphorylation Status of Ectopically Expressed Proteins in Early Xenopus Embryos Laura Hardwick and Anna Philpott 4. Analysis of Chromatin Binding of Ectopically Expressed Proteins in Early Xenopus Embryos Laura Hardwick and Anna Philpott 5. Raising Antibodies for Use in Xenopus Maya Z.
Piccinni and Matthew J. Guille 6. Assessing the Immune Response When Raising Antibodies for Use in Xenopus #105593 Maya Z. Piccinni and Matthew J. Guille 7. Confirming Antibody Specificity in Xenopus Sian A. Martin, Suzannah J. Page, Maya Z.
Piccinni, and Matthew J. Guille 8. Purifying Antibodies Raised against Xenopus Peptides Maya Z. Piccinni and Matthew J. Guille SECTION 7: XENOPUS EXTRACT SYSTEMS Editor: Rebecca Heald INTRODUCTION The Use of Cell-Free Xenopus Extracts to Investigate Cytoplasmic Events Romain Gibeaux and Rebecca Heald PROTOCOLS 1. Preparation of Cellular Extracts from Xenopus Eggs and Embryos Matthew C. Good and Rebecca Heald 2. Robustly Cycling Xenopus laevis Cell-Free Extracts in Teflon Chambers Jeremy B.
Chang and James E. Ferrell, Jr. 3. Protein Immunodepletion and Complementation in Xenopus laevis Egg Extracts Christopher Jenness, David Wynne, and Hironori Funabiki 4. Assembly of Spindles and Asters in Xenopus Egg Extracts Christine M. Field and Timothy J. Mitchison 5. Dissecting Protein Complexes in Branching Microtubule Nucleation using Meiotic Xenopus Egg Extracts Jae-Geun Song and Sabine Petry 6.
Filopodia-Like Structure Formation from Xenopus Egg Extracts Helen Mary Fox and Jennifer Louise Gallop 7. Endoplasmic Reticulum Network Formation with Xenopus Egg Extracts Songyu Wang, Fabian B. Romano, and Tom A. Rapoport 8. Microfluidic Encapsulation of Demembranated Sperm Nuclei in Xenopus Egg Extracts John Oakey and Jesse C. Gatlin 9. Calculating the Degradation Rate of Individual Proteins using Xenopus Extract Systems Gary S. McDowell and Anna Philpott 10.
Assessing Ubiquitylation of Individual Proteins Using Xenopus Extract Systems Gary S. McDowell and Anna Philpott SECTION 8: CHROMOSOME DYNAMICS IN VITRO Editor: Rebecca Heald INTRODUCTION Reconstituting Nuclear and Chromosome Dynamics Using Xenopus Extracts Susannah Rankin PROTOCOLS 1. Isolation and Demembranation of Xenopus Sperm Nuclei James W. Hazel and Jesse C. Gatlin 2. Chromosome Cohesion and Condensation in Xenopus Egg Extracts Eullia M.L. da Silva and Susannah Rankin 3.
Chromatin Characterization in Xenopus laevis Cell-Free Egg Extracts and Embryos Wei-Lin Wang, Takashi Onikubo, and David Shechter 4. Centromere and Kinetochore Assembly in Xenopus laevis Egg Extract Julio Servin and Aaron F. Straight 5. Analysis of Mitotic Checkpoint Function in Xenopus Egg Extracts Yinghui Mao 6. Nucleus Assembly and Import in Xenopus laevis Egg Extract Pan Chen and Daniel L. Levy 7. Extracts for Analysis of DNA Replication in a Nucleus-Free System Justin Lee Sparks and Johannes Walter SECTION 9: IMAGING Editors: Lance A. Davidson and Laura Anne Lowery INTRODUCTION Imaging Methods in Xenopus Cells, Embryos, and Tadpoles Lance A.
Davidson and Laura Anne Lowery PROTOCOLS 1 Whole-Mount RNA In Situ Hybridization and Immunofluorescence of Xenopus Embryos and Tadpoles Helen Rankin Willsey 2. Cryosectioning and Immunostaining of Xenopus Embryonic Tissues Olga Ossipova and Sergei Y. Sokol 3 Live Imaging of Cytoskeletal Dynamics in Embryonic Xenopus laevis Growth Cones and Neura.