Planarian Regeneration

Description

This volume explores the various facets of planaria as a biomedical model system and discusses techniques used to study the fascinating biology of these animals. The chapters in this book are divided into two parts: Part One looks at the biodiversity of planarian species, the molecular orchestration of regeneration, ecology of planarians in their natural habitats and their history as lab models. Part Two talks about experimental protocols for studying planarians, ranging from the establishment of a planarian research colony, to RNA and DNA extraction techniques, all the way to single stem cell transplantations or metabolomics analysis. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

 Comprehensive and cutting-edge, Planarian Regeneration: Methods and Protocols is a valuable resource for both newcomers to the field and experts within established planarian laboratories. Front Matter ....Pages i-xi Planarian Diversity and Phylogeny (Ronald Sluys, Marta Riutort)....Pages 1-56 Stem Cells, Patterning and Regeneration in Planarians: Self-Organization at the Organismal Scale (Jochen C. Rink)....Pages 57-172 The Ecology of Freshwater Planarians (Miquel Vila-Farré, Jochen C. Rink)....Pages 173-205 Planarians and the History of Animal Regeneration: Paradigm Shifts and Key Concepts in Biology (Sarah A. Elliott, Alejandro Sánchez Alvarado)....Pages 207-239 Culturing Planarians in the Laboratory (M. Shane Merryman, Alejandro Sánchez Alvarado, James C. Jenkin)....Pages 241-258 Total RNA Isolation from Planarian Tissues (Shang-Yun Liu, Jochen C. Rink)....Pages 259-265 Small- and Large-Scale High Molecular Weight Genomic DNA Extraction from Planarians (Markus A. Grohme, Miquel Vila-Farré, Jochen C. Rink)....Pages 267-275 Planarian High Molecular Weight DNA Isolation by Spooling (Shasha Zhang, Alejandro Sánchez Alvarado)....Pages 277-284 Basic Histological Techniques for Planarians (Leigh Winsor, Ronald Sluys)....Pages 285-351 Fixation, Processing, and Immunofluorescent Labeling of Whole Mount Planarians (David J. Forsthoefel, Kelly G. Ross, Phillip A. Newmark, Ricardo M. Zayas)....Pages 353-366 Immunohistochemistry on Paraffin-Embedded Planarian Tissue Sections (Teresa Adell, Sara Barberán, Miquel Sureda-Gómez, María Almuedo-Castillo, Nidia de Sousa, Francesc Cebrià)....Pages 367-378 Whole-Mount In Situ Hybridization of Planarians (Ryan S. King, Phillip A. Newmark)....Pages 379-392 RNA In Situ Hybridization on Planarian Paraffin Sections (Jordi Solana)....Pages 393-404 General Principles of Planarian Embryogenesis and Its Analysis by In Situ Hybridization and Immunohistochemistry Methods (José María Martín-Durán)....Pages 405-421 Whole-Mount BrdU Staining with Fluorescence In Situ Hybridization in Planarians (Li-Chun Cheng, Alejandro Sánchez Alvarado)....Pages 423-434 Detection of Apoptotic Cells in Planarians by Whole-Mount TUNEL (Brad Stubenhaus, Jason Pellettieri)....Pages 435-444 Systemic RNA Interference in Planarians by Feeding of dsRNA Containing Bacteria (Carolyn E. Adler, Alejandro Sánchez Alvarado)....Pages 445-454 RNA Interference in Planarians: Feeding and Injection of Synthetic dsRNA (Norito Shibata, Kiyokazu Agata)....Pages 455-466 A Subtractive FACS Method for Isolation of Planarian Stem Cells and Neural Cells (Tetsutaro Hayashi, Kiyokazu Agata)....Pages 467-478 Clonal Analysis of Planarian Stem Cells by Subtotal Irradiation and Single-Cell Transplantation (Irving E. Wang, Daniel E. Wagner, Peter W. Reddien)....Pages 479-495 Tissue Transplantations in Planarians (José I. Rojo-Laguna, Emili Saló)....Pages 497-505 Live Imaging of Planaria (Wei Shen, Yun Shen, Yun Wah Lam, Danny Chan)....Pages 507-518 Processing Schmidtea mediterranea for Transmission Electron Microscopy: Classical and Microwave Techniques (John L. Brubacher, Ana P. Vieira, Juliette Azimzadeh)....Pages 519-538 Tissue Extracts for Quantitative Mass Spectrometry of Planarian Proteins Using SILAC (Alexander Böser, Hannes C. A. Drexler, Kerstin Bartscherer)....Pages 539-553 Quantification of Neurotransmitters from Intact and Regenerating Planarians Using UHPLC-MS/SRM Method (Kannan Rangiah, Dasaradhi Palakodeti)....Pages 555-570 Back Matter ....Pages 571-573