Dictyostelium discoideum Protocols

Dictyostelium discoideum Protocols

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Dictyostelium discoideum is a simple but fascinating eukaryotic microorg- ism, whose natural habitat is deciduous forest soil and decaying leaves, where the amoebae feed on bacteria and grow as independent single cells. Exhaustion of the bacterial food source triggers a developmental program, in which up to 100,000 cells aggregate by chemotaxis towards cAMP. Morphogenesis and cell different- tion then culminate in the production of spores enabling the organism to survive unfavorable conditions. Dictyostelium offers unique advantages for studying f- damental cellular processes with the aid of powerful molecular genetic, bioche- cal, and cell biological tools. These processes include signal transduction, chemotaxis, cell motility, cytokinesis, phagocytosis, and aspects of development such as cell sorting, pattern formation and cell type differentiation. Recently, D- tyostelium was also described as a suitable host for pathogenic bacteria in which one can conveniently study the process of infection. In addition, Dictyostelium has many of the experimental conveniences of Saccharomyces cerevisiae and is pr- ably the best experimentally manipulatable protozoan, providing insight into this diverse group of organisms, which includes some of the most dangerous human parasites. The recent completion of the Dictyostelium genome sequencing project strengthens the position of D. discoideum as a model organism. The completed genome sequence and other valuable community resources constitute the source for basic biological and biomedical research and for genome-wide analyses.
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Product details

  • Hardback | 564 pages
  • 154.9 x 231.1 x 33mm | 907.2g
  • Totowa, NJ, United States
  • English
  • 2006 ed.
  • 186 Illustrations, black and white; XVI, 564 p. 186 illus.
  • 1588296237
  • 9781588296238

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Dictyostelium discoideum Protocols presents a comprehensive collection of cutting-edge molecular biological protocols for studying D. discoideum, with emphasis on its utility in the study of fundamental cellular processes including signal transduction, chemotaxis, cell motility, cytokinesis, phagocytosis, and aspects of development such as cell sorting, pattern formation, and cell type differentiation. Written by experts in the field, this important guide provides easy access to current methods and techniques for the study of D. discoideum. For the newcomer to the field, it offers a complete and inclusive introduction to this important model organism. The book is divided into four main parts, which include an introduction to the organism that provides integral community resources and genome-wide approaches; basic methods and available molecular genetic techniques for study of the organism; imaging and localization methods; and a discussion of D. discoideum, with emphasis on its unique advantages as a model system.
This in-depth guide provides a complete introduction to and thorough handbook for the study of D. discoideum, including a discussion of the implications of the recently completed Dictyostelium genome project in strengthening the position of D. discoideum as a model organism for studying fundamental cell processes and aspects of development. Researchers at all levels of experience will benefit from the presentations throughout of the most current, useful, and innovative techniques for the study of D. discoideum from leading Dictyostelium scientists.
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Table of contents

Part I. The Organism, Community Resources, and Genome-wide Studies

The Secret Lives of Dictyostelium
Richard H. Kessin

The Genome of Dictyostelium discoideum
Adam Kuspa and William F. Loomis

The cDNA Sequencing Project
Hideko Urushihara, Takahiro Morio, and Yoshimasa Tanaka

dictyBase and the Dicty Stock Center
Petra Fey, Pascale Gaudet, Karen E. Pilcher, Jakob Franke, and Rex L. Chisholm

Analysis of Gene Expression Using cDNA Microarrays
Marcel Kaul and Ludwig Eichinger

Proteomic Analysis of Dictyostelium discoideum
Udo Roth, Stefan Muller, and Franz-Georg Hanisch

Part II. Basic Methods and Molecular Genetic Techniques

Cultivation, Spore Production, and Mating
Hideko Urushihara

Parasexual Genetics Using Axenic Cells
Jason King and Robert Insall

Slug Phototaxis, Thermotaxis, and Spontaneous Turning Behavior
Paul R. Fisher and Sarah J. Annesley

Purification Techniques of Subcellular Compartments for Analytical and Preparative Purposes
Laurence Aubry and Gerard Klein

Generation of Multiple Knockout Mutants Using the Cre-loxP System
Alan R. Kimmel and Jan Faix

Restriction Enzyme-Mediated Integration (REMI) Mutagenesis
Adam Kuspa

RNA Interference and Antisense-Mediated Gene Silencing in Dictyostelium
Markus Kuhlmann, Blagovesta Popova, and Wolfgang Nellen

Part III. Imaging and Localization Methods

Application of Fluorescent Protein Tags as Reporters in Live-Cell Imaging Studies
Annette Muller-Taubenberger

Investigating Gene Expression: In Situ Hybridization and Reporter Genes
Ricardo Escalante and Leandro Sastre

Application of 2D and 3D DIAS to Motion Analysis of Live Cells in Transmission and Confocal Microscopy Imaging
Deborah Wessels, Spencer Kuhl, and David R. Soll

UsingQuantitative Fluorescence Microscopy and FRET Imaging to Measure Spatiotemporal Signaling Events in Single Living Cells
Xuehua Xu, Joseph A. Brzostowski, and Tian Jin

Visualizing Signaling and Cell Movement During the Multicellular Stages of Dictyostelium Development
Dirk Dormann and Cornelis J. Weijer

Under-Agarose Chemotaxis of Dictyostelium discoideum
David Woznica and David A. Knecht

Optimized Fixation and Immunofluorescence Staining Methods for Dictyostelium Cells
Monica Hagedorn, Eva M. Neuhaus, and Thierry Soldati

Cryofixation Methods for Ultrastructural Studies of Dictyostelium discoideum
Mark J. Grimson and Richard L. Blanton

Part IV. Dictyostelium as Model Organism

Analysis of Signal Transduction: Formation of cAMP, cGMP, and Ins(1,4,5)P3 In Vivo and In Vitro
Peter J. M. Van Haastert

Assaying Chemotaxis of Dictyostelium Cells
Michelle C. Mendoza and Richard A. Firtel

Characterization of Cross-Linked Actin Filament Gels and Bundles Using Birefringence and Polarized Light Scattering
Ruth Furukawa and Marcus Fechheimer

Quantitative and Microscopic Methods for Studying the Endocytic Pathway
Francisco Rivero and Markus Maniak

Preparation of Intact, Highly Purified Phagosomes from Dictyostelium
Daniel Gotthardt, Regis Dieckmann, Vincent Blancheteau, Claudia Kistler, Frank Reichardt, and Thierry Soldati

Assaying Cell-Cell Adhesion
Salvatore Bozzaro

Periodic Activation of ERK2 and Partial Involvement of G Protein in ERK2 Activation by cAMP in Dictyostelium Cells
Mineko Maeda

An Improved Method for Dictyostelium Centrosome Isolation
Irene Schulz, Yvonne Reinders, Albert Sickmann, and Ralph Graf

Epigenetics in Dictyostelium
Markus Kaller, Wolfgang Nellen, and Jonathan R. Chubb

Dictyostelium discoideum as a Model to Study
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