Dictyostelium discoideum Protocols / Edition 1

by Ludwig Eichinger

Overview

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.

Product Details

ISBN-13:	9781617376771
Publisher:	Springer-Verlag New York, LLC
Publication date:	11/19/2010
Series:	Methods in Molecular Biology , #346
Edition description:	Softcover reprint of hardcover 1st ed. 2006
Pages:	564
Product dimensions:	5.98(w) x 9.02(h) x 0.35(d)

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 Müller, 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 Gérard 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 Müller-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

Using Quantitative 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, Régis 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 Gräf

Epigenetics in Dictyostelium

Markus Kaller, Wolfgang Nellen, and Jonathan R. Chubb

Dictyostelium discoideum as a Model to Study Host-Pathogen Interactions

Can Ünal and Michael Steinert

Pharmacogenetics: Defining the Genetic Basis of Drug Action and Inositol Trisphosphate Analysis

Kathryn E. Adley, Melanie Keim, and Robin S. B.Williams

How to Assess and Study Cell Death in Dictyostelium discoideum

Artemis Kosta, Catherine Laporte, David Lam, Emilie Tresse, Marie-Françoise Luciani, and Pierre Golstein

Index