Dictyostelium discoideum protocols pdf

Methods in Molecular Biology, Vol. Thus, based on these premises, should result nearly obvious the biological reasons setting forth an increasingly use of this model organism for the study of cell motility, chemotaxis, phagocytosis, endocytic vesicle traffic, cell adhesion, pattern formation, caspase-independent cell death, and, more recently, autophagy and social evolution ; what can result less obvious is the fact that now-a-days Dictyostelium is being increasingly used in the study of human diseases host-pathogen interactions, microbial infections, mitochondrial diseases, pharmacogenetic studies eventhough we are speaking of a nonmammalian model, as clearly stated by Salvatore Bozzaro Dept.

All of the studies using Dictyostelium got a powerful acceleration from the publication of its genome sequence because from that moment large-scale methods are amenable to many studies. Chapter 1 — 6 illustrate the many resources available to the Dictyostelium scientific community, particularly well describing the vaste opportunities provided by the dictyBase and the Dicty stock center.

In chapters 7 — 11 genome-wide analysis are described including a chapter devoted to the pharmacogenetics of cisplatin, and other anticancer drugs, resistance given particular attention to the role played by the sphingolipid metabolism. Chapters 12 — 20 illustrate a panoply of methods ranging from molecular genetics to biophysical techniques passing through the more traditional, but so useful, biochemical and cell biological techniques: the use of the Cre-loxP system to generate multiple Knock-out and Knock-in targeted loci is fascinating.

Dictyostelium discoideum is a well-establish eukaryotic model organism that offers unique advantages for studying fundamental cellular processes, including signal transduction, random and directed cell motility, cytokinesis, endocytosis and vesicle transport and development. Dictyostelium is also increasingly used for the investigation of human disease genes and the crosstalk between host and pathogen.

The availability of the genome sequence at a mouse click together with a whole range of supporting information and resources along with a powerful armoury of molecular genetics techniques have considerably enhanced the experimental attractiveness of D.

The second edition of Dictyostelium discoideum Protocols incorporates the most recent developments in a number of fields. The book is divided into four parts. The first part provides an introduction to the amoebozoa and community resources. The second part presents large-scale analysis methods made possible by the completion of the Dictyostelium genome sequence.

The third part is dedicated to molecular genetics techniques, cell biological, biochemical and biophysical methods. The chapters in the fourth part describe the use of Dictyostelium as a model system to study vesicle formation, trafficking and infection by bacterial pathogens.

Authoritative and practical, Dictyostelium discoideum Protocols, Second Edition seeks to aid scientists in the application of innovating techniques to study a range of fundamental biological processes in this attractive model organism. Skip to main content Skip to table of contents. Advertisement Hide. This service is more advanced with JavaScript available. Dictyostelium discoideum Protocols. Editors view affiliations Ludwig Eichinger Francisco Rivero. Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.

Front Matter Pages i-xii. The Amoebozoa. In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript. Dictyostelium amoebae have been used as a host model to measure virulence of a wide range of bacterial pathogens. The simple protocol described here takes advantage of the ability of Dictyostelium to grow and form plaques on a lawn of nonpathogenic bacteria but not on virulent bacteria.

This assay can be modulated to measure the virulence of different bacterial pathogens. By adjusting various parameters such as cell numbers or media, a more quantitative measure of bacterial virulence can also be obtained. The entire procedure takes about 5 h to compete, and up to 2 further weeks may be required for plaques to form on the bacterial lawn.

Casadevall, A. Host—pathogen interactions: redefining the basic concepts of virulence and pathogenicity. Hilbi, H. Environmental predators as models for bacterial pathogenesis. Kurz, C. Infection in a dish: high-throughput analyses of bacterial pathogenesis. Cosson, P. Eat, kill or die: when amoeba meets bacteria, Curr. Fey, P. Protocols for growth and development of Dictyostelium discoideum. Eichinger, L.

The genome of the social amoeba Dictyostelium discoideum. Nature , 43 Sussman, R. Cultivation of Dictyostelium discoideum in axenic medium. Watts, D. Growth of myxameobae of the cellular slime mould Dictyostelium discoideum in axenic culture. Pseudomonas aeruginosa virulence analyzed in a Dictyostelium discoideum host system. Alibaud, L. Pseudomonas aeruginosa virulence genes identified in a Dictyostelium host model.

Cell Microbiol. Froquet, R.