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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.
Inhaltsverzeichnis
Planarian Diversity and Phylogeny.- Stem Cells, Patterning, and Regeneration in Planarians-Self-Organization at the Organismal Scale.- The Ecology of Freshwater Planarians.- Planarians and the History of Animal Regeneration: Paradigm Shifts and Key Concepts in Biology.- Culturing Planarians in the Laboratory.- Total RNA Isolation from Planarian Tissues.- Small- and Large-Scale High Molecular Weight Genomic DNA Extraction from Planarians.- Planarian High Molecular Weight DNA Isolation by Spooling.- Basic Histological Techniques for Planarians.- Fixation, Processing, and Immunofluorescent Labeling of Whole Mount Planarians.- Immunohistochemistry on Paraffin-Embedded Planarian Tissue Sections.- Whole-Mount In Situ Hybridization of Planarians.- RNA In Situ Hybridization on Planarian Paraffin Sections.- General Principles of Planarian Embryogenesis and Its Analysis by In Situ Hybridization and Immunohistochemistry Methods.- Whole-Mount BrdU Staining with Fluorescence In Situ Hybridization in Planarians.- Detection of Apoptotic Cells in Planarians by Whole-Mount TUNEL.- Systemic RNA Interference in Planarians by Feeding of dsRNA Containing Bacteria.- RNA Interference in Planarians: Feeding and Injection of Synthetic dsRNA.- A Subtractive FACS Method for Isolation of Planarian Stem Cells and Neural Cells.- Clonal Analysis of Planarian Stem Cells by Subtotal Irradiation and Single-Cell Transplantation.- Tissue Transplantations in Planarians.- Live Imaging Planaria.- Processing Schmidtea mediterranea for Transmission Electron Microscopy: Classical and Microwave Techniques.- Tissue Extracts for Quantitative Mass Spectrometry of Planarian Proteins Using SILAC.- Quantification of Neurotransmitters from Intact and Regenerating Planarians Using UHPLC-MS/SRM Method.
Über den Autor / die Autorin
Jochen Rink, PhD
Group Leader
Max Planck Institute of Molecular Cell Biology and Genetics
Pfotenhauerstrasse 108, 01307 Dresden, Germany
Tel: +49-(0)351-210-2435
Email: rink@mpi-cbg.de
Zusammenfassung
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.
